Disease Information

General Information of the Disease (ID: DIS00146)

• Name: Astrocytoma
• ICD: ICD-11: 2F36

Type(s) of Resistant Mechanism of This Disease

  ADTT: Aberration of the Drug's Therapeutic Target

  DISM: Drug Inactivation by Structure Modification

  IDUE: Irregularity in Drug Uptake and Drug Efflux

  UAPP: Unusual Activation of Pro-survival Pathway

Drug Resistance Data Categorized by Drug

Approved Drug(s) 12 drug(s) in total

Amodiaquine

Drug Resistance Data Categorized by Their Corresponding Mechanisms

Irregularity in Drug Uptake and Drug Efflux (IDUE)

Key Molecule: Chloroquine resistance transporter (CRT) [1]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.K76T
• Resistant Drug: Amodiaquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: MIP probes and PCR sequencing assay
• Experiment for Drug Resistance: SYBR Green I detection assay
• Mechanism Description: Increasingly, molecular genetic markers for antimalarial drug resistance have been identified, an advance that facilitates the monitoring of the emergence and spread of resistance. Currently, reliable molecular markers are available for P. falciparum resistance to artemisinins (mutations in the propeller region of Pfkelch), sulfadoxine-pyrimethamine (mutations in the dihydrofolate reductase [PfDHFR] and dihydropteroate synthase [PfDHPS] genes), mefloquine (MQ) (amplification of the multidrug resistance-1 gene [PfMDR1]), and piperaquine (amplification of PfPlasmepsin2/3 and specific mutations in the P. falciparum chloroquine resistance transporter gene.

Key Molecule: Multidrug resistance protein 1 (ABCB1) [1]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.N86Y
• Resistant Drug: Amodiaquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: MIP probes and PCR sequencing assay
• Experiment for Drug Resistance: SYBR Green I detection assay
• Mechanism Description: Increasingly, molecular genetic markers for antimalarial drug resistance have been identified, an advance that facilitates the monitoring of the emergence and spread of resistance. Currently, reliable molecular markers are available for P. falciparum resistance to artemisinins (mutations in the propeller region of Pfkelch), sulfadoxine-pyrimethamine (mutations in the dihydrofolate reductase [PfDHFR] and dihydropteroate synthase [PfDHPS] genes), mefloquine (MQ) (amplification of the multidrug resistance-1 gene [PfMDR1]), and piperaquine (amplification of PfPlasmepsin2/3 and specific mutations in the P. falciparum chloroquine resistance transporter gene.

Drug Sensitivity Data Categorized by Their Corresponding Mechanisms

Irregularity in Drug Uptake and Drug Efflux (IDUE)

Key Molecule: Chloroquine resistance transporter (CRT) [2]

• Sensitive Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.C101F
• Sensitive Drug: Amodiaquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum asexual blood-stage parasites; 5833
• Experiment for Molecule Alteration: DNA clones asssay
• Experiment for Drug Resistance: SYBR Green I detection assay
• Mechanism Description: This mutation (C101F) also reversed Dd2-mediated CQ resistance, sensitized parasites to amodiaquine, quinine, and artemisinin, and conferred amantadine and blasticidin resistance.

Artemisinin

Drug Sensitivity Data Categorized by Their Corresponding Mechanisms

Irregularity in Drug Uptake and Drug Efflux (IDUE)

Key Molecule: Chloroquine resistance transporter (CRT) [2]

• Sensitive Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.C101F
• Sensitive Drug: Artemisinin
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum asexual blood-stage parasites; 5833
• Experiment for Molecule Alteration: DNA clones asssay
• Experiment for Drug Resistance: SYBR Green I detection assay
• Mechanism Description: This mutation (C101F) also reversed Dd2-mediated CQ resistance, sensitized parasites to amodiaquine, quinine, and artemisinin, and conferred amantadine and blasticidin resistance.

Chloroquine

Drug Resistance Data Categorized by Their Corresponding Mechanisms

Irregularity in Drug Uptake and Drug Efflux (IDUE)

Key Molecule: Chloroquine resistance transporter (CRT) [3], [4], [5]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.H97Y
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: PacBio amplicon sequencing assay; Whole genome sequencing assay
• Experiment for Drug Resistance: Piperaquine susceptibility testing assay
• Mechanism Description: In parasites with single-copy pfpm2, those with the PfCRT F145I, G353V, or I218F mutations had a significantly greater log10-transformed piperaquine IC90 compared to Dd2 (linear regression; P <.0001, P =.0022, and P =.019, respectively), while other mutations did not show a significant difference in piperaquine IC90 compared to Dd2 (perhaps owing to smaller sample.

Key Molecule: Chloroquine resistance transporter (CRT) [6], [7], [8]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.K76T
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: MIP probes and PCR sequencing assay
• Experiment for Drug Resistance: SYBR Green I detection assay; [3H]-hypoxanthine assay
• Mechanism Description: Notably, the PfCRT Lys76Thr substitution was associated with significantly decreased susceptibility to chloroquine, monodesethylamodiaquine, and AQ-13; associations with other aminoquinolines were not conclusive.

Key Molecule: Multidrug resistance protein 1 (ABCB1) [1], [9], [10]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.N86Y
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: MIP probes and PCR sequencing assay
• Experiment for Drug Resistance: SYBR Green I detection assay
• Mechanism Description: Increasingly, molecular genetic markers for antimalarial drug resistance have been identified, an advance that facilitates the monitoring of the emergence and spread of resistance. Currently, reliable molecular markers are available for P. falciparum resistance to artemisinins (mutations in the propeller region of Pfkelch), sulfadoxine-pyrimethamine (mutations in the dihydrofolate reductase [PfDHFR] and dihydropteroate synthase [PfDHPS] genes), mefloquine (MQ) (amplification of the multidrug resistance-1 gene [PfMDR1]), and piperaquine (amplification of PfPlasmepsin2/3 and specific mutations in the P. falciparum chloroquine resistance transporter gene.

Key Molecule: Chloroquine resistance transporter (CRT) [4], [11], [12]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.C350R
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: Amino acid sequence alignments assay
• Experiment for Drug Resistance: Flow cytometry assay
• Mechanism Description: Functional studies on the newly emerging PfCRT F145I and C350R mutations, associated with decreased PPQ susceptibility in Asia and South America respectively reveal their ability to mediate PPQ transport in 7G8 variant proteins and to confer resistance in gene-edited parasites. The apparent dichotomy observed between CQ and PPQ for the F145I and C350R mutants, which evolved on CQ-R isoforms and caused CQ resensitization along with a gain of PPQ resistance, highlights the value of extending this research to rapidly emerging PfCRT mutations.

Key Molecule: Multidrug resistance protein 1 (ABCB1) [13], [14], [15]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.Y184F
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: Nested PCR; Sequence assay
• Experiment for Drug Resistance: [3H]-hypoxanthine assay
• Mechanism Description: Parasites with a chloroquine IC50 > 100 nM were significantly associated with PfCRT 97L and pfmdr1 184F, and a pfmdr1 copy number >= 4 was more common in those with a chloroquine IC50 <=100 nM.

Key Molecule: Chloroquine resistance transporter (CRT) [16], [17]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.M74I
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: Direct DNA sequencing method assay
• Mechanism Description: High prevalence of mutant Pfcrt genotypes associated with chloroquine resistance in Assam and Arunachal Pradesh, India. The k76T mutation was observed in 77.78% cases followed by M74I (61.11%), N75E (61.11%) and C72S (16.67%). Triple mutant allele M74I+N75E+k76T was found in 61.11% P. falciparum field isolates. Double mutant allele C72S+k76T was seen among 16.67% samples.

Key Molecule: Chloroquine resistance transporter (CRT) [16], [17]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.N75E
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: Direct DNA sequencing method assay
• Mechanism Description: High prevalence of mutant Pfcrt genotypes associated with chloroquine resistance in Assam and Arunachal Pradesh, India. The k76T mutation was observed in 77.78% cases followed by M74I (61.11%), N75E (61.11%) and C72S (16.67%). Triple mutant allele M74I+N75E+k76T was found in 61.11% P. falciparum field isolates. Double mutant allele C72S+k76T was seen among 16.67% samples.

Key Molecule: Chloroquine resistance transporter (CRT) [7], [17]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.A220S
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: [3H]-hypoxanthine assay
• Mechanism Description: Plasmodium falciparum chloroquine resistance (CQR) transporter protein (PfCRT) is the important key of CQR. There is a link between mutations in the gene pfcrt and resistance to chloroquine in P. falciparum. Almost all of the parasites characterized carried the previously reported mutations k76T, A220S, Q271E, N326S, I356T and R371I. On complete sequencing, isolates were identified with novel mutations at k76A and E198K.

Key Molecule: Chloroquine resistance transporter (CRT) [7], [17]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.I356T
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: [3H]-hypoxanthine assay
• Mechanism Description: Plasmodium falciparum chloroquine resistance (CQR) transporter protein (PfCRT) is the important key of CQR. There is a link between mutations in the gene pfcrt and resistance to chloroquine in P. falciparum. Almost all of the parasites characterized carried the previously reported mutations k76T, A220S, Q271E, N326S, I356T and R371I. On complete sequencing, isolates were identified with novel mutations at k76A and E198K.

Key Molecule: Chloroquine resistance transporter (CRT) [7], [17]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.N326S
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: [3H]-hypoxanthine assay
• Mechanism Description: Plasmodium falciparum chloroquine resistance (CQR) transporter protein (PfCRT) is the important key of CQR. There is a link between mutations in the gene pfcrt and resistance to chloroquine in P. falciparum. Almost all of the parasites characterized carried the previously reported mutations k76T, A220S, Q271E, N326S, I356T and R371I. On complete sequencing, isolates were identified with novel mutations at k76A and E198K.

Key Molecule: Chloroquine resistance transporter (CRT) [7], [17]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.R371I
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: [3H]-hypoxanthine assay
• Mechanism Description: Plasmodium falciparum chloroquine resistance (CQR) transporter protein (PfCRT) is the important key of CQR. There is a link between mutations in the gene pfcrt and resistance to chloroquine in P. falciparum. Almost all of the parasites characterized carried the previously reported mutations k76T, A220S, Q271E, N326S, I356T and R371I. On complete sequencing, isolates were identified with novel mutations at k76A and E198K.

Key Molecule: Chloroquine resistance transporter (CRT) [18]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.C101F+p.F145I+p.M343L+p.G353V+T93S+I218F
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum D10; 5833
• In Vitro Model: Plasmodium falciparum Dd2; 5833
• In Vitro Model: Plasmodium falciparum PfCRT; 5833
• Experiment for Molecule Alteration: Western blotting analysis
• Mechanism Description: Plasmodium falciparum parasites resistant to chloroquine, amodiaquine, or piperaquine harbor mutations in the P. falciparum chloroquine resistance transporter (PfCRT), a transporter resident on the digestive vacuole membrane that in its variant forms can transport these weak-base 4-aminoquinoline drugs out of this acidic organelle, thus preventing these drugs from binding heme and inhibiting its detoxification.

Key Molecule: Chloroquine resistance transporter (CRT) [19]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.C101
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum isolate 3D7; 5833
• In Vitro Model: Plasmodium falciparum isolate 7G8; 5833
• In Vitro Model: Plasmodium falciparum isolate Dd2; 5833
• Experiment for Molecule Alteration: Western blotting analysis
• Experiment for Drug Resistance: P. falciparum proliferation assay
• Mechanism Description: Drug-resistance-conferring mutations reduce both the peptide transport capacity and substrate range of PfCRT, explaining the impaired fitness of drug-resistant parasites. Two PfCRT mutations that arose separately under in vitro drug pressure (C101F and L272F) incur both a fitness cost and a monstrously swollen DV. Three laboratory-derived isoforms that cause a gross enlargement of the DV-L272F-PfCRT3D7, L272F-PfCRTDd2 and C101F-PfCRTDd2-displayed unusually low capacities for VF-6 tra.

Key Molecule: Chloroquine resistance transporter (CRT) [19]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.L272F
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum isolate 3D7; 5833
• In Vitro Model: Plasmodium falciparum isolate 7G8; 5833
• In Vitro Model: Plasmodium falciparum isolate Dd2; 5833
• Experiment for Molecule Alteration: Western blotting analysis
• Experiment for Drug Resistance: P. falciparum proliferation assay
• Mechanism Description: Drug-resistance-conferring mutations reduce both the peptide transport capacity and substrate range of PfCRT, explaining the impaired fitness of drug-resistant parasites. Two PfCRT mutations that arose separately under in vitro drug pressure (C101F and L272F) incur both a fitness cost and a monstrously swollen DV. Three laboratory-derived isoforms that cause a gross enlargement of the DV-L272F-PfCRT3D7, L272F-PfCRTDd2 and C101F-PfCRTDd2-displayed unusually low capacities for VF-6 tra.

Key Molecule: Chloroquine resistance transporter (CRT) [4]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.C101F
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum W2; 5833
• Experiment for Molecule Alteration: Pfcrt genotyping assay
• Experiment for Drug Resistance: HRP2 ELISA-based assay Malaria Ag Celisa kit assay
• Mechanism Description: In a context of dihydroartemisinin-piperaquine resistance in Cambodia and high prevalence of k13 C580Y mutation associated with artemisinin resistance, new pfcrt mutations (H97Y, M343L, and G353V) were revealed to induce in vitro piperaquine resistance. Treatment failures with dihydroartemisinin-piperaquine were associated with T93S, H97Y, F145I and I218F mutations in PfCRT and with plasmepsin 2/3 amplification in Cambodia, Thailand and Vietnam.

Key Molecule: Chloroquine resistance transporter (CRT) [4]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.F145I
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum W2; 5833
• Experiment for Molecule Alteration: Pfcrt genotyping assay
• Experiment for Drug Resistance: HRP2 ELISA-based assay Malaria Ag Celisa kit assay
• Mechanism Description: In a context of dihydroartemisinin-piperaquine resistance in Cambodia and high prevalence of k13 C580Y mutation associated with artemisinin resistance, new pfcrt mutations (H97Y, M343L, and G353V) were revealed to induce in vitro piperaquine resistance. Treatment failures with dihydroartemisinin-piperaquine were associated with T93S, H97Y, F145I and I218F mutations in PfCRT and with plasmepsin 2/3 amplification in Cambodia, Thailand and Vietnam.

Key Molecule: Chloroquine resistance transporter (CRT) [4]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.G353V
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum W2; 5833
• Experiment for Molecule Alteration: Pfcrt genotyping assay
• Experiment for Drug Resistance: HRP2 ELISA-based assay Malaria Ag Celisa kit assay
• Mechanism Description: In a context of dihydroartemisinin-piperaquine resistance in Cambodia and high prevalence of k13 C580Y mutation associated with artemisinin resistance, new pfcrt mutations (H97Y, M343L, and G353V) were revealed to induce in vitro piperaquine resistance. Treatment failures with dihydroartemisinin-piperaquine were associated with T93S, H97Y, F145I and I218F mutations in PfCRT and with plasmepsin 2/3 amplification in Cambodia, Thailand and Vietnam.

Key Molecule: Chloroquine resistance transporter (CRT) [4]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.M343L
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum W2; 5833
• Experiment for Molecule Alteration: Pfcrt genotyping assay
• Experiment for Drug Resistance: HRP2 ELISA-based assay Malaria Ag Celisa kit assay
• Mechanism Description: In a context of dihydroartemisinin-piperaquine resistance in Cambodia and high prevalence of k13 C580Y mutation associated with artemisinin resistance, new pfcrt mutations (H97Y, M343L, and G353V) were revealed to induce in vitro piperaquine resistance. Treatment failures with dihydroartemisinin-piperaquine were associated with T93S, H97Y, F145I and I218F mutations in PfCRT and with plasmepsin 2/3 amplification in Cambodia, Thailand and Vietnam.

Key Molecule: Chloroquine resistance transporter (CRT) [4]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.T93S
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum W2; 5833
• Experiment for Molecule Alteration: Pfcrt genotyping assay
• Experiment for Drug Resistance: HRP2 ELISA-based assay Malaria Ag Celisa kit assay
• Mechanism Description: In a context of dihydroartemisinin-piperaquine resistance in Cambodia and high prevalence of k13 C580Y mutation associated with artemisinin resistance, new pfcrt mutations (H97Y, M343L, and G353V) were revealed to induce in vitro piperaquine resistance. Treatment failures with dihydroartemisinin-piperaquine were associated with T93S, H97Y, F145I and I218F mutations in PfCRT and with plasmepsin 2/3 amplification in Cambodia, Thailand and Vietnam.

Key Molecule: Chloroquine resistance transporter (CRT) [17]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.C72S
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• Mechanism Description: Mutant PfCRT molecules have acquired the ability to expel CQ out of the DV. All CQR haplotypes carry the key k76T mutation that removes a positive charge in TM 1, suggesting a charge-dependent transport mechanism as CQ is di-protonated in the acidic DV10,13. The k76T mutation is always accompanied by additional mutations which may increase the CQR level and/or attenuate the fitness cost of resistance.

Key Molecule: Chloroquine resistance transporter (CRT) [17]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.I356L
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• Mechanism Description: Mutant PfCRT molecules have acquired the ability to expel CQ out of the DV. All CQR haplotypes carry the key k76T mutation that removes a positive charge in TM 1, suggesting a charge-dependent transport mechanism as CQ is di-protonated in the acidic DV10,13. The k76T mutation is always accompanied by additional mutations which may increase the CQR level and/or attenuate the fitness cost of resistance.

Key Molecule: Chloroquine resistance transporter (CRT) [17]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.N326D
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• Mechanism Description: Mutant PfCRT molecules have acquired the ability to expel CQ out of the DV. All CQR haplotypes carry the key k76T mutation that removes a positive charge in TM 1, suggesting a charge-dependent transport mechanism as CQ is di-protonated in the acidic DV10,13. The k76T mutation is always accompanied by additional mutations which may increase the CQR level and/or attenuate the fitness cost of resistance.

Key Molecule: Multidrug resistance protein 1 (ABCB1) [9]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.N86F
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Mechanism Description: Resistance to chloroquine (CQ) in P. falciparum parasites is predominantly linked to a single mutation in the P. falciparum transporter gene (Pfcrt) on chromosome 7, which encodes a protein localized on the parasite digestive vacuole (DV) membrane. The replacement of lysine (k) at position 76 to a threonine (T), i.e. the k76T mutation, has been established as the most important prognostic marker of treatment failure. Another point mutation N86Y in P. falciparum multidrug resistance gene 1 (Pfmdr1), on chromosome 5, that encodes a P-glycoprotein homologue and is located on the parasite DV membrane has also been implicated in CQ resistance.

Key Molecule: Chloroquine resistance transporter (CRT) [11]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.F145I
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: Amino acid sequence alignments assay
• Experiment for Drug Resistance: Flow cytometry assay
• Mechanism Description: Functional studies on the newly emerging PfCRT F145I and C350R mutations, associated with decreased PPQ susceptibility in Asia and South America respectively reveal their ability to mediate PPQ transport in 7G8 variant proteins and to confer resistance in gene-edited parasites. The apparent dichotomy observed between CQ and PPQ for the F145I and C350R mutants, which evolved on CQ-R isoforms and caused CQ resensitization along with a gain of PPQ resistance, highlights the value of extending this research to rapidly emerging PfCRT mutations.

Key Molecule: Chloroquine resistance transporter (CRT) [20]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Phosphorylation; Up-regulation
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: Western blotting analysis
• Experiment for Drug Resistance: SYBR Green I detection assay
• Mechanism Description: Phosphorylation at Ser-33 and Ser-411 of PfCRT of the chloroquine-resistant P. falciparum strain Dd2 and kinase inhibitors can sensitize drug responsiveness.

Key Molecule: Putative chloroquine resistance transporter (PVCRT) [21]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.K10 insertion (c.AAG)
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• Mechanism Description: Mutations in k10 insertion in the Pvcrt-o gene have been identified as a possible molecular marker of CQ resistance in P.vivax.

Key Molecule: Putative chloroquine resistance transporter (PVCRT) [21]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.K10 insertion (c.AAG)
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium vivax isolates; 5855
• Experiment for Molecule Alteration: Nested PCR
• Mechanism Description: Mutations in k10 insertion in the Pvcrt-o gene have been identified as a possible molecular marker of CQ resistance in P.vivax.

Key Molecule: Putative chloroquine resistance transporter (PVCRT) [22]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.S249P
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Yeast CH1305; 4932
• Experiment for Molecule Alteration: Western blotting analysis
• Experiment for Drug Resistance: Quantitative Growth Rate assay
• Mechanism Description: It is surprising then that the data presented here suggests a single mutation (S249P in PvCRT isoform CQR3) can increase CQ transport by 32% relative to wild type.

Key Molecule: Chloroquine resistance transporter (CRT) [16]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.C72S
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: Direct DNA sequencing method assay
• Mechanism Description: High prevalence of mutant Pfcrt genotypes associated with chloroquine resistance in Assam and Arunachal Pradesh, India. The k76T mutation was observed in 77.78% cases followed by M74I (61.11%), N75E (61.11%) and C72S (16.67%). Triple mutant allele M74I+N75E+k76T was found in 61.11% P. falciparum field isolates. Double mutant allele C72S+k76T was seen among 16.67% samples.

Key Molecule: Chloroquine resistance transporter (CRT) [16]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.C72S+p.K76T
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: Direct DNA sequencing method assay
• Mechanism Description: High prevalence of mutant Pfcrt genotypes associated with chloroquine resistance in Assam and Arunachal Pradesh, India. The k76T mutation was observed in 77.78% cases followed by M74I (61.11%), N75E (61.11%) and C72S (16.67%). Triple mutant allele M74I+N75E+k76T was found in 61.11% P. falciparum field isolates. Double mutant allele C72S+k76T was seen among 16.67% samples.

Key Molecule: Chloroquine resistance transporter (CRT) [16]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.M74I+p.N75E+p.K76T
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: Direct DNA sequencing method assay
• Mechanism Description: High prevalence of mutant Pfcrt genotypes associated with chloroquine resistance in Assam and Arunachal Pradesh, India. The k76T mutation was observed in 77.78% cases followed by M74I (61.11%), N75E (61.11%) and C72S (16.67%). Triple mutant allele M74I+N75E+k76T was found in 61.11% P. falciparum field isolates. Double mutant allele C72S+k76T was seen among 16.67% samples.

Key Molecule: Chloroquine resistance transporter (CRT) [23]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Chromosome variation; Dd2 genotype
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Drug Resistance: lactate dehydrogenase (pLDH) assay
• Mechanism Description: Chloroquine resistance-conferring isoforms of PfCRT reduced the susceptibility of the parasite to QC, MB, and AO. In chloroquine-resistant (but not chloroquine-sensitive) parasites, AO and QC increased the parasite's accumulation of, and susceptibility to, chloroquine. All 3 compounds were shown to bind to Pf.

Key Molecule: Chloroquine resistance transporter (CRT) [23]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Chromosome variation; K1 genotype
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Drug Resistance: lactate dehydrogenase (pLDH) assay
• Mechanism Description: Chloroquine resistance-conferring isoforms of PfCRT reduced the susceptibility of the parasite to QC, MB, and AO. In chloroquine-resistant (but not chloroquine-sensitive) parasites, AO and QC increased the parasite's accumulation of, and susceptibility to, chloroquine. All 3 compounds were shown to bind to Pf.

Key Molecule: Chloroquine resistance transporter (CRT) [24]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; Mutant pfcrt alleles PH1 and PH2
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Drug Resistance: [3H]-hypoxanthine assay
• Mechanism Description: Variant alleles from the Philippines (PH1 and PH2, which differ solely by the C72S mutation) both conferred a moderate gain of chloroquine resistance and a reduction in growth rates in vitro. Of the two, PH2 showed higher IC50 values, contrasting with reduced.

Key Molecule: Putative chloroquine resistance transporter (PVCRT) [25]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Expression; Up-regulation
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium vivax strains; 5855
• Mechanism Description: Patients with CQ-resistant P. vivax parasites presented a higher gene expression of pvcrt-o and pvmdr-1 at D0 and DR when compared to the susceptible group. For the CQR patients, median gene expression values at D0 and DR, presented 2.4 fold (95% CI: 0.96-7.1) and 6.1 fold (95% CI: 3.8-14.3) increase in pvcrt-o levels compared to the susceptible patients at D0 with 0.12 fold (95% CI: 0.034-0.324). Median gene expression for pvmdr-1 presented 2.0 fold (95% CI: 0.95-3.8) and 2.4 fold (95% CI: 0.53-9.1) increase levels at D0 and DR, for the CQR patients versus 0.288 fold (95% CI: 0.068-0.497) for the susceptible patients.

Key Molecule: Multidrug resistance protein 1 (ABCB1) [25]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Expression; Up-regulation
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium vivax strains; 5855
• Mechanism Description: Patients with CQ-resistant P. vivax parasites presented a higher gene expression of pvcrt-o and pvmdr-1 at D0 and DR when compared to the susceptible group. For the CQR patients, median gene expression values at D0 and DR, presented 2.4 fold (95% CI: 0.96-7.1) and 6.1 fold (95% CI: 3.8-14.3) increase in pvcrt-o levels compared to the susceptible patients at D0 with 0.12 fold (95% CI: 0.034-0.324). Median gene expression for pvmdr-1 presented 2.0 fold (95% CI: 0.95-3.8) and 2.4 fold (95% CI: 0.53-9.1) increase levels at D0 and DR, for the CQR patients versus 0.288 fold (95% CI: 0.068-0.497) for the susceptible patients.

Key Molecule: Chloroquine resistance transporter (CRT) [8]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.K76T
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum C-1Dd2; 5833
• In Vitro Model: Plasmodium falciparum C2GC03; 5833
• In Vitro Model: Plasmodium falciparum C3Dd2; 5833
• In Vitro Model: Plasmodium falciparum C67G8; 5833
• In Vitro Model: Plasmodium falciparum GC03; 5833
• In Vitro Model: Plasmodium falciparum T76k-1Dd2; 5833
• Experiment for Molecule Alteration: Western blotting analysis
• Experiment for Drug Resistance: [3H]-hypoxanthine assay
• Mechanism Description: The k76T mutation in PfCRT generates structural changes that are sufficient to allow GSH transport, but not CQ transport. mutant pfcrt allows enhanced transport of GSH into the parasite's DV. The elevated levels of GSH in the DV reduce the level of free heme available for CQ binding, which mediates the lower susceptibility to CQ in the PfCRT mutant parasites.

Key Molecule: Multidrug resistance protein 1 (ABCB1) [26]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.976F
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium vivax isolates; 5855
• Mechanism Description: In Southeast Asia the pvmdr1 976 F allele has been associated with reduced susceptibility to CQ. Finding the pvmdr1 976 F allele in 7/41 (17%) P. vivax might thus indicate a degree of CQ tolerance but probably not resistance in Honduras.

Key Molecule: Chloroquine resistance transporter (CRT) [13]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.H97L
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: Nested PCR
• Experiment for Drug Resistance: [3H]-hypoxanthine assay
• Mechanism Description: Parasites with a chloroquine IC50 > 100 nM were significantly associated with PfCRT 97L and pfmdr1 184F, and a pfmdr1 copy number >= 4 was more common in those with a chloroquine IC50 <=100 nM.

Key Molecule: Chloroquine resistance transporter (CRT) [7]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.E198K
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: [3H]-hypoxanthine assay
• Mechanism Description: Plasmodium falciparum chloroquine resistance (CQR) transporter protein (PfCRT) is the important key of CQR. There is a link between mutations in the gene pfcrt and resistance to chloroquine in P. falciparum. Almost all of the parasites characterized carried the previously reported mutations k76T, A220S, Q271E, N326S, I356T and R371I. On complete sequencing, isolates were identified with novel mutations at k76A and E198K.

Key Molecule: Chloroquine resistance transporter (CRT) [7]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.K76A
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: [3H]-hypoxanthine assay
• Mechanism Description: Plasmodium falciparum chloroquine resistance (CQR) transporter protein (PfCRT) is the important key of CQR. There is a link between mutations in the gene pfcrt and resistance to chloroquine in P. falciparum. Almost all of the parasites characterized carried the previously reported mutations k76T, A220S, Q271E, N326S, I356T and R371I. On complete sequencing, isolates were identified with novel mutations at k76A and E198K.

Key Molecule: Chloroquine resistance transporter (CRT) [7]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.Q271E
• Resistant Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: [3H]-hypoxanthine assay
• Mechanism Description: Plasmodium falciparum chloroquine resistance (CQR) transporter protein (PfCRT) is the important key of CQR. There is a link between mutations in the gene pfcrt and resistance to chloroquine in P. falciparum. Almost all of the parasites characterized carried the previously reported mutations k76T, A220S, Q271E, N326S, I356T and R371I. On complete sequencing, isolates were identified with novel mutations at k76A and E198K.

Drug Sensitivity Data Categorized by Their Corresponding Mechanisms

Irregularity in Drug Uptake and Drug Efflux (IDUE)

Key Molecule: Chloroquine resistance transporter (CRT) [27], [28]

• Sensitive Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Expression; Down-regulation
• Sensitive Drug: Chloroquine
• Experimental Note: Revealed Based on the Cell Line Data
• Cell Pathway Regulation: Cell proliferation; Inhibition; hsa05200
• In Vitro Model: Human foreskin fibroblasts; N.A.; Saccharomyces cerevisiae; N.A.
• In Vitro Model: Toxoplasma gondii Prudetaku80S/Luc; 1080348
• In Vivo Model: C57BL/6J mice xenograft model; Mus musculus
• Experiment for Molecule Alteration: Western blotting analysis
• Experiment for Drug Resistance: Tachyzoite plaque assay
• Mechanism Description: Rrecombinant PfCRT transports amino acids, small peptides, and chloroquine, suggests that PfCRT functions to transport products of hemoglobin digestion out of the digestive vacuole. TgCRT is also able to transport amino acids and small peptides out of the VAC. TgCRT-deficient tachyzoites also grow more slowly in vitro and are compromised in their ability to cause mortality in mice during acute infection, suggesting that an inability to transport digested material out of the VAC and into the parasite cytosol has a moderate effect on T. gondii tachyzoites.

Key Molecule: Chloroquine resistance transporter (CRT) [29], [30], [31]

• Sensitive Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Expression; Down-regulation
• Sensitive Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: Western blotting analysis
• Experiment for Drug Resistance: SYBR Green I detection assay; [3H]-hypoxanthine assay
• Mechanism Description: Phosphorylation at Ser-33 and Ser-411 of PfCRT of the chloroquine-resistant P. falciparum strain Dd2 and kinase inhibitors can sensitize drug responsiveness.

Key Molecule: Chloroquine resistance transporter (CRT) [2]

• Sensitive Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.C101F
• Sensitive Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum asexual blood-stage parasites; 5833
• Experiment for Molecule Alteration: DNA clones asssay
• Experiment for Drug Resistance: SYBR Green I detection assay
• Mechanism Description: This mutation (C101F) also reversed Dd2-mediated CQ resistance, sensitized parasites to amodiaquine, quinine, and artemisinin, and conferred amantadine and blasticidin resistance.

Key Molecule: Chloroquine resistance transporter (CRT) [32]

• Sensitive Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.C101F
• Sensitive Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum 3D7; 36329
• In Vitro Model: Plasmodium falciparum 3D7L272F mutant; 36329
• Experiment for Drug Resistance: SYBR Green I detection assay
• Mechanism Description: Carry PfCRT mutations (C101F or L272F), causing the development of enlarged food vacuoles. These parasites also have increased sensitivity to chloroquine and some other quinoline antimalarials. Furthermore, the introduction of the C101F or L272F mutation into a chloroquine-resistant variant of PfCRT reduced the ability of this protein to transport chloroquine by approximately 93 and 82% when expressed in Xenopus oocytes.

Key Molecule: Chloroquine resistance transporter (CRT) [32]

• Sensitive Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.L272F
• Sensitive Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum 3D7; 36329
• In Vitro Model: Plasmodium falciparum 3D7L272F mutant; 36329
• Experiment for Drug Resistance: SYBR Green I detection assay
• Mechanism Description: Carry PfCRT mutations (C101F or L272F), causing the development of enlarged food vacuoles. These parasites also have increased sensitivity to chloroquine and some other quinoline antimalarials. Furthermore, the introduction of the C101F or L272F mutation into a chloroquine-resistant variant of PfCRT reduced the ability of this protein to transport chloroquine by approximately 93 and 82% when expressed in Xenopus oocytes.

Key Molecule: Chloroquine resistance transporter (CRT) [30]

• Sensitive Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.S163R
• Sensitive Drug: Chloroquine
• Experimental Note: Revealed Based on the Cell Line Data
• In Vitro Model: Plasmodium falciparum D10; 5833
• In Vitro Model: Plasmodium falciparum Dd2; 5833
• In Vitro Model: Plasmodium falciparum PfCRT; 5833
• Mechanism Description: T76k and S163R mutations in PfCRTCQR restore CQ sensitivity to CQR parasites. The introduction of either one of these changes to PfCRTCQR, each of which entailed the addition of a positive charge to the putative substrate-binding site of the protein, resulted in the loss of CQ transport activity.

Key Molecule: Chloroquine resistance transporter (CRT) [30]

• Sensitive Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.T76K
• Sensitive Drug: Chloroquine
• Experimental Note: Revealed Based on the Cell Line Data
• In Vitro Model: Plasmodium falciparum D10; 5833
• In Vitro Model: Plasmodium falciparum Dd2; 5833
• In Vitro Model: Plasmodium falciparum PfCRT; 5833
• Mechanism Description: T76k and S163R mutations in PfCRTCQR restore CQ sensitivity to CQR parasites. The introduction of either one of these changes to PfCRTCQR, each of which entailed the addition of a positive charge to the putative substrate-binding site of the protein, resulted in the loss of CQ transport activity.

Key Molecule: ABC transporter (ABCT) [29]

• Sensitive Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Expression; Down-regulation
• Sensitive Drug: Chloroquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Mechanism Description: Phenothiazine drugs (chlor-promazine, trifluoperazine, prochlorperazine, methotrime-prazin or fluphenazin) can enhance in vitro the potency of CQ against P. falciparum CQ-resistant strains. phenothiazines may exert their CQ resistance reversal activity by interacting with Pgh1. Combinations of chlorpromazine or prochlorperazine with CQ confirm the reversal effect of these drugs on the CQ resistance as showed by cures obtained in Aotus monkeys infected with CQ-resistant P. falciparum.

Chlorpheniramine

Drug Resistance Data Categorized by Their Corresponding Mechanisms

Irregularity in Drug Uptake and Drug Efflux (IDUE)

Key Molecule: Chloroquine resistance transporter (CRT) [33]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.K76N
• Resistant Drug: Chlorpheniramine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum; 5833
• Experiment for Drug Resistance: Malaria SYBR Green I-based fluorescence (MSF) method assay
• Mechanism Description: Mutations within PfCRT, particularly changes from a charged amino acid residue (lysine, k76) to an uncharged residue (such as threonine [76T], asparagine [76N], or isoleucine [76I]), seem to be important not only in the acquisition of resistance to quinoline antimalarials (e.g., by allowing efflux of diprotic CQ), but also in the mechanism of resistance reversal actions for chemosens.

Key Molecule: Chloroquine resistance transporter (CRT) [33]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.K76T
• Resistant Drug: Chlorpheniramine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum; 5833
• Experiment for Drug Resistance: Malaria SYBR Green I-based fluorescence (MSF) method assay
• Mechanism Description: Mutations within PfCRT, particularly changes from a charged amino acid residue (lysine, k76) to an uncharged residue (such as threonine [76T], asparagine [76N], or isoleucine [76I]), seem to be important not only in the acquisition of resistance to quinoline antimalarials (e.g., by allowing efflux of diprotic CQ), but also in the mechanism of resistance reversal actions for chemosens.

Glutathione

Drug Resistance Data Categorized by Their Corresponding Mechanisms

Irregularity in Drug Uptake and Drug Efflux (IDUE)

Key Molecule: Chloroquine resistance transporter (CRT) [8]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.K76T
• Resistant Drug: Glutathione
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum C-1Dd2; 5833
• In Vitro Model: Plasmodium falciparum C2GC03; 5833
• In Vitro Model: Plasmodium falciparum C3Dd2; 5833
• In Vitro Model: Plasmodium falciparum C67G8; 5833
• In Vitro Model: Plasmodium falciparum GC03; 5833
• In Vitro Model: Plasmodium falciparum T76k-1Dd2; 5833
• Experiment for Molecule Alteration: Western blotting analysis
• Experiment for Drug Resistance: [3H]-hypoxanthine assay
• Mechanism Description: The k76T mutation in PfCRT generates structural changes that are sufficient to allow GSH transport, but not CQ transport. mutant pfcrt allows enhanced transport of GSH into the parasite's DV. The elevated levels of GSH in the DV reduce the level of free heme available for CQ binding, which mediates the lower susceptibility to CQ in the PfCRT mutant parasites.

Drug Sensitivity Data Categorized by Their Corresponding Mechanisms

Drug Inactivation by Structure Modification (DISM)

Key Molecule: Gamma-glutamylcysteine synthetase (GGCS) [34]

• Sensitive Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Expression; Down-regulation
• Sensitive Drug: Glutathione
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium berghei ANkA 2.34; 5823
• In Vitro Model: Plasmodium berghei pbggcs-ko; 5821
• In Vitro Model: Plasmodium berghei pbggcs-oe; 5821
• In Vivo Model: Swiss albino CD-1 female mice xenograft model; Mus musculus
• Experiment for Molecule Alteration: Western blotting analysis
• Experiment for Drug Resistance: In vivo drug suppressive test assay
• Mechanism Description: To analyze the role of GSH levels in CQ and ART resistance, we generated transgenic Plasmodium berghei parasites either deficient in or overexpressing the gamma-glutamylcysteine synthetase gene (pbggcs) encoding the rate-limiting enzyme in GSH biosynthesis. These lines produce either lower (pbggcs-ko) or higher (pbggcs-oe) levels of GSH than wild type parasites. Recrudescence assays after the parasites have been exposed to a sub-lethal dose of ART showed that parasites with low levels of GSH are more sensitive to ART tre.

Halofantrine

Drug Sensitivity Data Categorized by Their Corresponding Mechanisms

Irregularity in Drug Uptake and Drug Efflux (IDUE)

Key Molecule: Multidrug resistance protein 1 (ABCB1) [35]

• Sensitive Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Expression; Down-regulation
• Sensitive Drug: Halofantrine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Drug Resistance: SYBR Green I detection assay
• Mechanism Description: The presence of non-toxic concentrations of Mk571 sensitized both chloroquine-sensitive and -resistant parasites to mefloquine and halofantrine, likely by competing against PfMDR1-mediated sequestering of the drugs into the DV compartment and away from the drugs' cytosolic targets.

Mefloquine

Drug Resistance Data Categorized by Their Corresponding Mechanisms

Irregularity in Drug Uptake and Drug Efflux (IDUE)

Key Molecule: Multidrug resistance protein 1 (ABCB1) [36], [1]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Expression; Up-regulation
• Resistant Drug: Mefloquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: Western blotting analysis
• Experiment for Drug Resistance: SYBR Green I detection assay
• Mechanism Description: Increasingly, molecular genetic markers for antimalarial drug resistance have been identified, an advance that facilitates the monitoring of the emergence and spread of resistance. Currently, reliable molecular markers are available for P. falciparum resistance to artemisinins (mutations in the propeller region of Pfkelch), sulfadoxine-pyrimethamine (mutations in the dihydrofolate reductase [PfDHFR] and dihydropteroate synthase [PfDHPS] genes), mefloquine (MQ) (amplification of the multidrug resistance-1 gene [PfMDR1]), and piperaquine (amplification of PfPlasmepsin2/3 and specific mutations in the P. falciparum chloroquine resistance transporter gene.

Drug Sensitivity Data Categorized by Their Corresponding Mechanisms

Irregularity in Drug Uptake and Drug Efflux (IDUE)

Key Molecule: Chloroquine resistance transporter (CRT) [1]

• Sensitive Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.N86Y
• Sensitive Drug: Mefloquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: MIP probes and PCR sequencing assay
• Experiment for Drug Resistance: SYBR Green I detection assay
• Mechanism Description: Despite the availability of few mutant parasites for comparison, the PfMDR1 Asn86Tyr substitution appeared to be associated with increased susceptibility to lumefantrine and mefloquine, as seen prev.

Key Molecule: Multidrug resistance protein 1 (ABCB1) [37]

• Sensitive Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.N86+p.Y184
• Sensitive Drug: Mefloquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum isolates; 5833
• Experiment for Molecule Alteration: Quantitative trait loci (QTL) assay
• Experiment for Drug Resistance: SYBR Green I detection assay
• Mechanism Description: By QTL analysis, lumefantrine and mefloquine phenotypes mapped to a chromosome 5 region containing codon polymorphisms N86Y and Y184F in the P. falciparum multidrug resistance 1 protein.

Key Molecule: Chloroquine resistance transporter (CRT) [10]

• Sensitive Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.K76T
• Sensitive Drug: Mefloquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: Nested PCR
• Mechanism Description: Both in vitro and molecular surveillance studies have associated CQ resistance mainly with the pfcrt 76T allele, but also with pfmdr1 86Y and 184F alleles. Pfcrt 76T and pfmdr1 86Y mutant alleles have also been reported to decrease P. falciparum susceptibility to amodiaquine but increase parasite sensitivity to dihydroartemisinin, lumefantrine and mefl.

Key Molecule: Multidrug resistance protein 1 (ABCB1) [10]

• Sensitive Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.Y184F
• Sensitive Drug: Mefloquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: Nested PCR
• Mechanism Description: Both in vitro and molecular surveillance studies have associated CQ resistance mainly with the pfcrt 76T allele, but also with pfmdr1 86Y and 184F alleles. Pfcrt 76T and pfmdr1 86Y mutant alleles have also been reported to decrease P. falciparum susceptibility to amodiaquine but increase parasite sensitivity to dihydroartemisinin, lumefantrine and mefl.

Piperaquine

Drug Resistance Data Categorized by Their Corresponding Mechanisms

Irregularity in Drug Uptake and Drug Efflux (IDUE)

Key Molecule: Chloroquine resistance transporter (CRT) [3], [4], [5]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.H97Y
• Resistant Drug: Piperaquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: MIP probes and PCR sequencing assay
• Experiment for Drug Resistance: SYBR Green I detection assay
• Mechanism Description: In contrast, gene-edited parasites with PfCRT H97Y, F145I, M343L, or G353V mutations are resistant to piperaquine in vitro.

Key Molecule: Chloroquine resistance transporter (CRT) [3], [1]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.F145I
• Resistant Drug: Piperaquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: MIP probes and PCR sequencing assay
• Experiment for Drug Resistance: SYBR Green I detection assay
• Mechanism Description: In contrast, gene-edited parasites with PfCRT H97Y, F145I, M343L, or G353V mutations are resistant to piperaquine in vitro.

Key Molecule: Chloroquine resistance transporter (CRT) [3], [1]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.G353V
• Resistant Drug: Piperaquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: MIP probes and PCR sequencing assay
• Experiment for Drug Resistance: SYBR Green I detection assay
• Mechanism Description: In contrast, gene-edited parasites with PfCRT H97Y, F145I, M343L, or G353V mutations are resistant to piperaquine in vitro.

Key Molecule: Chloroquine resistance transporter (CRT) [3], [1]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.M343L
• Resistant Drug: Piperaquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: MIP probes and PCR sequencing assay
• Experiment for Drug Resistance: SYBR Green I detection assay
• Mechanism Description: In contrast, gene-edited parasites with PfCRT H97Y, F145I, M343L, or G353V mutations are resistant to piperaquine in vitro.

Key Molecule: Chloroquine resistance transporter (CRT) [5]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.F145I + p.G353V+ p.I218F
• Resistant Drug: Piperaquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: PacBio amplicon sequencing assay; Whole genome sequencing assay
• Experiment for Drug Resistance: Piperaquine susceptibility testing assay
• Mechanism Description: In parasites with single-copy pfpm2, those with the PfCRT F145I, G353V, or I218F mutations had a significantly greater log10-transformed piperaquine IC90 compared to Dd2 (linear regression; P <.0001, P =.0022, and P =.019, respectively), while other mutations did not show a significant difference in piperaquine IC90 compared to Dd2 (perhaps owing to smaller sample.

Key Molecule: Chloroquine resistance transporter (CRT) [5]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation + Chromosome variation; PfCRT p.F145I+p.G353V+p.I218F + Haplotype
• Resistant Drug: Piperaquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: PacBio amplicon sequencing assay; Whole genome sequencing assay
• Experiment for Drug Resistance: Piperaquine susceptibility testing assay
• Mechanism Description: Parasites with the Dd2 haplotype and pfpm2 amplification had significantly greater mean log10-transformed piperaquine IC90 compared to Dd2 parasites without pfpm2 amplification (t test, P =.0079). In parasites with newly emerged PfCRT mutations, mean log10-transformed piperaquine IC90 was not significantly different between parasites with or without pfpm2 amplification.

Key Molecule: Chloroquine resistance transporter (CRT) [38]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.T93S+p.H97Y+p.F145I+p.I218F
• Resistant Drug: Piperaquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: Western blotting analysis
• Experiment for Drug Resistance: Piperaquine survival assay
• Mechanism Description: The characterization of culture-adapted isolates revealed that the presence of novel pfcrt mutations (T93S, H97Y, F145I, and I218F) with E415G-Exo mutation can confer PPQ-resistance, in the absence of pfpm2 amplification.

Key Molecule: Putative chloroquine resistance transporter (PVCRT) [12]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.C350R
• Resistant Drug: Piperaquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Yeast strain CH1305; 4932
• Experiment for Molecule Alteration: Western blotting analysis
• Mechanism Description: At 300 uM PPQ, C350R/7G8 PfCRT shows a 3.9-fold increased rate of PPQ transport relative to that of 7G8 PfCRT, and F145I/Dd2 PfCRT shows a 2.7-fold increased rate of PPQ transport relative to that of Dd2.

Key Molecule: Chloroquine resistance transporter (CRT) [2]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.C101F
• Resistant Drug: Piperaquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum asexual blood-stage parasites; 5833
• Experiment for Molecule Alteration: DNA clones asssay
• Experiment for Drug Resistance: SYBR Green I detection assay
• Mechanism Description: Addition of the C101F mutation to the chloroquine (CQ) resistance-conferring PfCRT Dd2 isoform common to Asia can confer PPQ resistance to cultured parasites. Resistance was demonstrated as significantly higher PPQ concentrations causing 90% inhibition of parasite growth (IC90) or 50% parasite killing.

Unusual Activation of Pro-survival Pathway (UAPP)

Key Molecule: Plasmepsin II (PMII) [5]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation + Chromosome variation; PfCRT p.F145I+p.G353V+p.I218F + pfpm2 Amplification
• Resistant Drug: Piperaquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: PacBio amplicon sequencing assay; Whole genome sequencing assay
• Experiment for Drug Resistance: Piperaquine susceptibility testing assay
• Mechanism Description: Parasites with the Dd2 haplotype and pfpm2 amplification had significantly greater mean log10-transformed piperaquine IC90 compared to Dd2 parasites without pfpm2 amplification (t test, P =.0079). In parasites with newly emerged PfCRT mutations, mean log10-transformed piperaquine IC90 was not significantly different between parasites with or without pfpm2 amplification.

Key Molecule: Plasmepsin II (PMII) [5]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Chromosome variation; pfpm2 Amplification+Haplotype
• Resistant Drug: Piperaquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: PacBio amplicon sequencing assay; Whole genome sequencing assay
• Experiment for Drug Resistance: Piperaquine susceptibility testing assay
• Mechanism Description: Parasites with the Dd2 haplotype and pfpm2 amplification had significantly greater mean log10-transformed piperaquine IC90 compared to Dd2 parasites without pfpm2 amplification (t test, P?=?.0079).

Pyrimethamine/Sulfadoxine

Drug Resistance Data Categorized by Their Corresponding Mechanisms

Aberration of the Drug's Therapeutic Target (ADTT)

Key Molecule: Dihydrofolate reductase (DHFR) [39]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.C59R
• Resistant Drug: Pyrimethamine/Sulfadoxine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: PCR
• Mechanism Description: The quintuple mutant of pfdhfr (S108N, N51I and C59R) and pfdhps (A437G and k540E) were associated with a high relative risk of treatment failure, and this haplotype was suggested as a relevant molecular marker for failure of SP treatment in uncomplicated P. falciparum.

Key Molecule: Dihydrofolate reductase (DHFR) [39]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.C59R+I164L
• Resistant Drug: Pyrimethamine/Sulfadoxine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: PCR
• Mechanism Description: The quintuple mutant of pfdhfr (S108N, N51I and C59R) and pfdhps (A437G and k540E) were associated with a high relative risk of treatment failure, and this haplotype was suggested as a relevant molecular marker for failure of SP treatment in uncomplicated P. falciparum.

Key Molecule: Dihydrofolate reductase (DHFR) [39]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.C59R+S108N
• Resistant Drug: Pyrimethamine/Sulfadoxine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: PCR
• Mechanism Description: The quintuple mutant of pfdhfr (S108N, N51I and C59R) and pfdhps (A437G and k540E) were associated with a high relative risk of treatment failure, and this haplotype was suggested as a relevant molecular marker for failure of SP treatment in uncomplicated P. falciparum.

Key Molecule: Dihydrofolate reductase (DHFR) [39]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.C59R+S108N+I164L
• Resistant Drug: Pyrimethamine/Sulfadoxine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: PCR
• Mechanism Description: The quintuple mutant of pfdhfr (S108N, N51I and C59R) and pfdhps (A437G and k540E) were associated with a high relative risk of treatment failure, and this haplotype was suggested as a relevant molecular marker for failure of SP treatment in uncomplicated P. falciparum.

Key Molecule: Dihydrofolate reductase (DHFR) [39]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.N51I+C59R
• Resistant Drug: Pyrimethamine/Sulfadoxine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: PCR
• Mechanism Description: The quintuple mutant of pfdhfr (S108N, N51I and C59R) and pfdhps (A437G and k540E) were associated with a high relative risk of treatment failure, and this haplotype was suggested as a relevant molecular marker for failure of SP treatment in uncomplicated P. falciparum.

Key Molecule: Dihydrofolate reductase (DHFR) [39]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.N51I+C59R+I164L
• Resistant Drug: Pyrimethamine/Sulfadoxine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: PCR
• Mechanism Description: The quintuple mutant of pfdhfr (S108N, N51I and C59R) and pfdhps (A437G and k540E) were associated with a high relative risk of treatment failure, and this haplotype was suggested as a relevant molecular marker for failure of SP treatment in uncomplicated P. falciparum.

Key Molecule: Dihydrofolate reductase (DHFR) [39]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.N51I+C59R+S108N
• Resistant Drug: Pyrimethamine/Sulfadoxine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: PCR
• Mechanism Description: The quintuple mutant of pfdhfr (S108N, N51I and C59R) and pfdhps (A437G and k540E) were associated with a high relative risk of treatment failure, and this haplotype was suggested as a relevant molecular marker for failure of SP treatment in uncomplicated P. falciparum.

Key Molecule: Dihydrofolate reductase (DHFR) [39]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.N51I+C59R+S108N+I164L
• Resistant Drug: Pyrimethamine/Sulfadoxine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: PCR
• Mechanism Description: The quintuple mutant of pfdhfr (S108N, N51I and C59R) and pfdhps (A437G and k540E) were associated with a high relative risk of treatment failure, and this haplotype was suggested as a relevant molecular marker for failure of SP treatment in uncomplicated P. falciparum.

Key Molecule: Dihydrofolate reductase (DHFR) [39]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.A437G
• Resistant Drug: Pyrimethamine/Sulfadoxine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: PCR
• Mechanism Description: The quintuple mutant of pfdhfr (S108N, N51I and C59R) and pfdhps (A437G and k540E) were associated with a high relative risk of treatment failure, and this haplotype was suggested as a relevant molecular marker for failure of SP treatment in uncomplicated P. falciparum.

Pyronaridine

Drug Resistance Data Categorized by Their Corresponding Mechanisms

Irregularity in Drug Uptake and Drug Efflux (IDUE)

Key Molecule: Chloroquine resistance transporter (CRT) [40]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.K76T
• Resistant Drug: Pyronaridine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Drug Resistance: [3H]-hypoxanthine assay
• Mechanism Description: The pyronaridine IC50 (inhibitory concentration 50 %) ranged from 0.55 to 80.0 nM. Ex vivo responses to pyronaridine were significantly associated with the k76T mutation (p-value = 0.020). The reduced susceptibility to pyronaridine, defined as IC50 > 60 nM, was significantly associated with the k76T mutation (p-value = 0.004).

Quinine

Drug Resistance Data Categorized by Their Corresponding Mechanisms

Irregularity in Drug Uptake and Drug Efflux (IDUE)

Key Molecule: Chloroquine resistance transporter (CRT) [41], [42]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.76T
• Resistant Drug: Quinine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: PCR; Genotypic characterization assay
• Mechanism Description: Pfcrt is involved in the transport of quinine and that SNPs in pfcrt, including 76T, decrease P. falciparum susceptibility to quinine.

Key Molecule: Multidrug resistance protein 1 (ABCB1) [43]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.M908L
• Resistant Drug: Quinine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium vivax isolates; 5855
• Experiment for Molecule Alteration: In vitro drug assay
• Experiment for Drug Resistance: Analysis of genetic polymorphisms assay
• Mechanism Description: The pvmdr1 M908L substitutions in pvmdr1 in our samples was associated with reduced sensitivity to chloroquine, mefloquine, pyronaridine, piperaquine, quinine, artesunate and dihydroartem.

Key Molecule: Chloroquine resistance transporter (CRT) [4]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.I356T
• Resistant Drug: Quinine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Drug Resistance: Malaria Ag Celisa kit assay
• Mechanism Description: The mutation I356T, identified in 54.7% (n = 326) of the African isolates, was significantly associated with reduced susceptibility to quinine (p < 0.02) and increased susceptibility to mefloquine.

Key Molecule: Chloroquine resistance transporter (CRT) [20]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Phosphorylation; Up-regulation
• Resistant Drug: Quinine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: Western blotting analysis
• Experiment for Drug Resistance: SYBR Green I detection assay
• Mechanism Description: Phosphorylation of Ser-33 augments the level of PfCRT-conferred resistance to the antimalarial drugs chloroquine and quinine via stimulation of the transport velocity.

Key Molecule: Multidrug resistance protein 1 (ABCB1) [41]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.184F
• Resistant Drug: Quinine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: Genotypic characterization assay
• Experiment for Drug Resistance: SYBR Green I detection assay
• Mechanism Description: Eighty-two percent of parasites resistant to quinine carried mutant alleles at these codons (Pfmdr1-86Y, Pfmdr1-184F, and Pfcrt-76T), whereas 74% of parasites susceptible to quinine carried the wild-type allele (Pfmdr1-N86, Pfmdr1-Y184, and Pfcrt-k76, respect.

Key Molecule: Multidrug resistance protein 1 (ABCB1) [41]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.86Y
• Resistant Drug: Quinine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: Genotypic characterization assay
• Experiment for Drug Resistance: SYBR Green I detection assay
• Mechanism Description: Eighty-two percent of parasites resistant to quinine carried mutant alleles at these codons (Pfmdr1-86Y, Pfmdr1-184F, and Pfcrt-76T), whereas 74% of parasites susceptible to quinine carried the wild-type allele (Pfmdr1-N86, Pfmdr1-Y184, and Pfcrt-k76, respect.

Key Molecule: Na+/H+ exchanger-1 (PFNHE1) [41]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation + Chromosome variation; ms4760+ 3 DNNND repeats
• Resistant Drug: Quinine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: Genotypic characterization assay
• Experiment for Drug Resistance: SYBR Green I detection assay
• Mechanism Description: Eighty-two percent of parasites resistant to quinine carried mutant alleles at these codons (Pfmdr1-86Y, Pfmdr1-184F, and Pfcrt-76T), whereas 74% of parasites susceptible to quinine carried the wild-type allele (Pfmdr1-N86, Pfmdr1-Y184, and Pfcrt-k76, respect.

Drug Sensitivity Data Categorized by Their Corresponding Mechanisms

Irregularity in Drug Uptake and Drug Efflux (IDUE)

Key Molecule: Multidrug resistance protein 1 (ABCB1) [44], [45]

• Sensitive Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.184F
• Sensitive Drug: Quinine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: Genotypic characterization assay
• Experiment for Drug Resistance: [3H]-hypoxanthine assay; In vitro sensitivity assay
• Mechanism Description: 86Y allele exhibited significantly increased QN sensitivity compared with the wild-type counterpart. The parasites with the pfmdr1 184F allele exhibited approximately twice less susceptible to QN than the parasites with the pfmd.

Key Molecule: Multidrug resistance protein 1 (ABCB1) [44], [45]

• Sensitive Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.86Y
• Sensitive Drug: Quinine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: Genotypic characterization assay
• Experiment for Drug Resistance: [3H]-hypoxanthine assay; In vitro sensitivity assay
• Mechanism Description: 86Y allele exhibited significantly increased QN sensitivity compared with the wild-type counterpart. The parasites with the pfmdr1 184F allele exhibited approximately twice less susceptible to QN than the parasites with the pfmd.

Key Molecule: Chloroquine resistance transporter (CRT) [46], [47]

• Sensitive Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.K76I
• Sensitive Drug: Quinine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: Sequence assay
• Experiment for Drug Resistance: SYBR Green I detection assay
• Mechanism Description: In addition to producing CQ resistance in P. falciparum, a novel PfCRT k76I mutation resulted in a dramatic increase in QN susceptibility, reversing the normally observed potency order of QD > QN.

Key Molecule: Chloroquine resistance transporter (CRT) [38]

• Sensitive Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.T93S+p.H97Y+p.F145I+p.I218F
• Sensitive Drug: Quinine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: Western blotting analysis
• Experiment for Drug Resistance: Drug combination assay
• Mechanism Description: The presence of novel pfcrt mutations (T93S, H97Y, F145I, and I218F) with E415G-Exo mutation can confer PPQ-resistance, in the absence of pfpm2 amplification. In vitro testing of PPQ resistant parasites demonstrated a bimodal dose-response, the existence of a swollen digestive vacuole phenotype, and an increased susceptibility to quinine, chloroquine, mefloquine and lumefa.

Key Molecule: Chloroquine resistance transporter (CRT) [20]

• Sensitive Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.S33A
• Sensitive Drug: Quinine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: Western blotting analysis
• Experiment for Drug Resistance: SYBR Green I detection assay
• Mechanism Description: Substituting Ser-33 with alanine reduced chloroquine and quinine resistance by 50% compared with the parental P. falciparum strain Dd2, whereas the phosphomimetic amino acid aspartic acid could fully and glutamic acid could partially reconstitute the level of chloroquine/quinine resistance.

Key Molecule: Chloroquine resistance transporter (CRT) [2]

• Sensitive Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.C101F
• Sensitive Drug: Quinine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum asexual blood-stage parasites; 5833
• Experiment for Molecule Alteration: DNA clones asssay
• Experiment for Drug Resistance: SYBR Green I detection assay
• Mechanism Description: This mutation (C101F) also reversed Dd2-mediated CQ resistance, sensitized parasites to amodiaquine, quinine, and artemisinin, and conferred amantadine and blasticidin resistance.

Key Molecule: Chloroquine resistance transporter (CRT) [48]

• Sensitive Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Expression; Down-regulation
• Sensitive Drug: Quinine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Mechanism Description: This study describes the activities of a series of dimeric quinine compounds. These agents were found to be the most potent inhibitors of PfCRTCQR described to date with IC50 values between 1 and 5 M but are not themselves substrates of the transporter.

Sulphadoxine

Drug Resistance Data Categorized by Their Corresponding Mechanisms

Aberration of the Drug's Therapeutic Target (ADTT)

Key Molecule: Dihydrofolate reductase (DHFR) [49]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.N51I+p.C59R+p.S108N
• Resistant Drug: Sulphadoxine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: PCR
• Mechanism Description: The high prevalence of the pfdhfr 108N (99%) and 51I + 108N/59R + 108N (92%) in our study indicate that decreased susceptibility to sulphadoxine-pyrimethamine is widespread in Pakistan. However, only seven patients had infections with the triple pfdhfr resistance associated haplotype and only one patient was infected with P. falciparum that had the quintuple pfdhfr + pfdhps haplotype associated with high grade sulphadoxine-pyrimethamine resistance. These results indicate that high grade resistance to sulphadoxine-pyrimethamine is not wide.

Key Molecule: Dihydrofolate reductase (DHFR) [49]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.G437A+p.E540K
• Resistant Drug: Sulphadoxine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: PCR
• Mechanism Description: The high prevalence of the pfdhfr 108N (99%) and 51I + 108N/59R + 108N (92%) in our study indicate that decreased susceptibility to sulphadoxine-pyrimethamine is widespread in Pakistan. However, only seven patients had infections with the triple pfdhfr resistance associated haplotype and only one patient was infected with P. falciparum that had the quintuple pfdhfr + pfdhps haplotype associated with high grade sulphadoxine-pyrimethamine resistance. These results indicate that high grade resistance to sulphadoxine-pyrimethamine is not wide.

Key Molecule: Dihydrofolate reductase (DHFR) [49]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.N51I+p.C59R+p.S108N
• Resistant Drug: Sulphadoxine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: PCR
• Mechanism Description: The high prevalence of the pfdhfr 108N (99%) and 51I + 108N/59R + 108N (92%) in our study indicate that decreased susceptibility to sulphadoxine-pyrimethamine is widespread in Pakistan. However, only seven patients had infections with the triple pfdhfr resistance associated haplotype and only one patient was infected with P. falciparum that had the quintuple pfdhfr + pfdhps haplotype associated with high grade sulphadoxine-pyrimethamine resistance. These results indicate that high grade resistance to sulphadoxine-pyrimethamine is not wide.

Clinical Trial Drug(s) 2 drug(s) in total

AQ-13

Drug Resistance Data Categorized by Their Corresponding Mechanisms

Irregularity in Drug Uptake and Drug Efflux (IDUE)

Key Molecule: Chloroquine resistance transporter (CRT) [50]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.K76T
• Resistant Drug: AQ-13
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: MIP probes and PCR sequencing assay
• Experiment for Drug Resistance: SYBR Green I detection assay
• Mechanism Description: Notably, the PfCRT Lys76Thr substitution was associated with significantly decreased susceptibility to chloroquine, monodesethylamodiaquine, and AQ-13; associations with other aminoquinolines were not conclusive.

Lumefantrine

Drug Resistance Data Categorized by Their Corresponding Mechanisms

Irregularity in Drug Uptake and Drug Efflux (IDUE)

Key Molecule: Chloroquine resistance transporter (CRT) [37]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.N326S+p.I356T
• Resistant Drug: Lumefantrine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: Quantitative trait loci (QTL) assay
• Experiment for Drug Resistance: SYBR Green I detection assay
• Mechanism Description: Comparisons of the MEF and HLF responses showed that the Cambodian 803 line, as for LUM, was less susceptible than Ghanaian GB4 to these drugs: the geometric mean EC50s of 803 relative to GB4 were 2.9-fold greater with MEF and 4.6-fold greater with HLF, whereas these were 2.0-fold greater with CQ and 1.7-fold reduced w.

Key Molecule: Multidrug resistance protein 1 (ABCB1) [36]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Expression; Up-regulation
• Resistant Drug: Lumefantrine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Drug Resistance: HRP-2 ELISA assay
• Mechanism Description: Isolates with multiple pfmdr1 copies had significantly higher IC50s against OZ78, OZ277, MQ, and LUM. In contrast, no significant differences in IC50s between isolates with single and multiple pfmdr1 copy numbers were observed for the other test compounds.

Drug Sensitivity Data Categorized by Their Corresponding Mechanisms

Irregularity in Drug Uptake and Drug Efflux (IDUE)

Key Molecule: Multidrug resistance protein 1 (ABCB1) [1]

• Sensitive Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.N86Y
• Sensitive Drug: Lumefantrine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: MIP probes and PCR sequencing assay
• Experiment for Drug Resistance: SYBR Green I detection assay
• Mechanism Description: Despite the availability of few mutant parasites for comparison, the PfMDR1 Asn86Tyr substitution appeared to be associated with increased susceptibility to lumefantrine and mefloquine, as seen prev.

Key Molecule: Multidrug resistance protein 1 (ABCB1) [37]

• Sensitive Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.N86+p.Y184
• Sensitive Drug: Lumefantrine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum isolates; 5833
• Experiment for Molecule Alteration: Quantitative trait loci (QTL) assay
• Experiment for Drug Resistance: SYBR Green I detection assay
• Mechanism Description: The geometric mean LUM EC50 of 803 was 5.8-fold greater than GB4 (3.21 nM, 95% Confidence Interval 2.80-3.66 nM vs. 0.55 nM, 95% CI 0.46-0.67 nM, respectively). The Cambodian 803 line, as for LUM, was less susceptible than Ghanaian GB4 to these drugs: the geometric mean EC50s of 803 relative to GB4 were 2.9-fold greater with MEF and 4.6-fold greater with HLF, whereas these were 2.0-fold greater with CQ and 1.7-fold reduced with DHA.

Key Molecule: Chloroquine resistance transporter (CRT) [10]

• Sensitive Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.K76T
• Sensitive Drug: Lumefantrine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: Nested PCR
• Mechanism Description: Both in vitro and molecular surveillance studies have associated CQ resistance mainly with the pfcrt 76T allele, but also with pfmdr1 86Y and 184F alleles. Pfcrt 76T and pfmdr1 86Y mutant alleles have also been reported to decrease P. falciparum susceptibility to amodiaquine but increase parasite sensitivity to dihydroartemisinin, lumefantrine and mefl.

Key Molecule: Multidrug resistance protein 1 (ABCB1) [10]

• Sensitive Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.N86Y
• Sensitive Drug: Lumefantrine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: Nested PCR
• Mechanism Description: Both in vitro and molecular surveillance studies have associated CQ resistance mainly with the pfcrt 76T allele, but also with pfmdr1 86Y and 184F alleles. Pfcrt 76T and pfmdr1 86Y mutant alleles have also been reported to decrease P. falciparum susceptibility to amodiaquine but increase parasite sensitivity to dihydroartemisinin, lumefantrine and mefl.

Investigative Drug(s) 2 drug(s) in total

Desethylamodiaquine

Drug Resistance Data Categorized by Their Corresponding Mechanisms

Irregularity in Drug Uptake and Drug Efflux (IDUE)

Key Molecule: Chloroquine resistance transporter (CRT) [50]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.K76T
• Resistant Drug: Desethylamodiaquine
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: MIP probes and PCR sequencing assay
• Experiment for Drug Resistance: SYBR Green I detection assay
• Mechanism Description: Notably, the PfCRT Lys76Thr substitution was associated with significantly decreased susceptibility to chloroquine, monodesethylamodiaquine, and AQ-13; associations with other aminoquinolines were not conclusive.

Dihydroartemisinin

Drug Resistance Data Categorized by Their Corresponding Mechanisms

Irregularity in Drug Uptake and Drug Efflux (IDUE)

Key Molecule: Multidrug resistance protein 1 (ABCB1) [43]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.M908L
• Resistant Drug: Dihydroartemisinin
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium vivax isolates; 5855
• Experiment for Drug Resistance: In vitro drug assay
• Mechanism Description: Studies of genetic polymorphisms in two candidate genes of drug resistance (pvmdr1 and pvcrt-o) of the P. vivax isolates from this area and found association between the M908L substitution in pvmdr1 with reduced sensitivities to and chloroquine, mefloquine, pyronaridine, piperaquine, quinine, artesunate and dihydroartem.

Key Molecule: Chloroquine resistance transporter (CRT) [51]

• Resistant Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.F145I
• Resistant Drug: Dihydroartemisinin
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Mechanism Description: The PfCRT 145I mutation was only observed in parasites with amplified plasmepsin II/III copy number, suggesting that perhaps in nature this mutation has only occurred or only attains high frequency on a background of amplified plasmepsin II/III. Moreover, the mean piperaquine IC90 was greater in parasites with both amplified plasmepsin II/III and PfCRT 145I compared with parasites with just amplified plasmepsin II/III, suggesting that 145I results in an additional resistance effect beyond that caused by amplified plasmepsin.

Drug Sensitivity Data Categorized by Their Corresponding Mechanisms

Irregularity in Drug Uptake and Drug Efflux (IDUE)

Key Molecule: Chloroquine resistance transporter (CRT) [10]

• Sensitive Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.K76T
• Sensitive Drug: Dihydroartemisinin
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: Nested PCR
• Mechanism Description: Both in vitro and molecular surveillance studies have associated CQ resistance mainly with the pfcrt 76T allele, but also with pfmdr1 86Y and 184F alleles. Pfcrt 76T and pfmdr1 86Y mutant alleles have also been reported to decrease P. falciparum susceptibility to amodiaquine but increase parasite sensitivity to dihydroartemisinin, lumefantrine and mefl.

Key Molecule: Multidrug resistance protein 1 (ABCB1) [10]

• Sensitive Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.N86Y
• Sensitive Drug: Dihydroartemisinin
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: Nested PCR
• Mechanism Description: Both in vitro and molecular surveillance studies have associated CQ resistance mainly with the pfcrt 76T allele, but also with pfmdr1 86Y and 184F alleles. Pfcrt 76T and pfmdr1 86Y mutant alleles have also been reported to decrease P. falciparum susceptibility to amodiaquine but increase parasite sensitivity to dihydroartemisinin, lumefantrine and mefl.

Key Molecule: Multidrug resistance protein 1 (ABCB1) [52]

• Sensitive Disease: Malaria [ICD-11: 1F45.0]
• Molecule Alteration: Missense mutation; p.N86Y
• Sensitive Drug: Dihydroartemisinin
• Experimental Note: Discovered Using In-vivo Testing Model
• In Vitro Model: Plasmodium falciparum strains; 5833
• Experiment for Molecule Alteration: DNA assay
• Experiment for Drug Resistance: SYBR Green I detection assay
• Mechanism Description: The most striking phenotype observed with the replacement of N86Y with the wild-type N86 residue was a significant increase in the IC50 and IC90 values for LMF, MFQ and DHA. In the case of DHA, the change to N86 resulted in -1.5-fold increased IC50 values in both backgrounds.

References

• Ref 1: Evolution of Multidrug Resistance in Plasmodium falciparum: a Longitudinal Study of Genetic Resistance Markers in the Greater Mekong Subregion. Antimicrob Agents Chemother. 2021 Nov 17;65(12):e0112121. doi: 10.1128/AAC.01121-21. Epub 2021 Sep 13.
• Ref 2: A Variant PfCRT Isoform Can Contribute to Plasmodium falciparum Resistance to the First-Line Partner Drug Piperaquine. mBio. 2017 May 9;8(3):e00303-17. doi: 10.1128/mBio.00303-17.
• Ref 3: Emerging Southeast Asian PfCRT mutations confer Plasmodium falciparum resistance to the first-line antimalarial piperaquine. Nat Commun. 2018 Aug 17;9(1):3314. doi: 10.1038/s41467-018-05652-0.
• Ref 4: Prevalence of mutations in the Plasmodium falciparum chloroquine resistance transporter, PfCRT, and association with ex vivo susceptibility to common anti-malarial drugs against African Plasmodium falciparum isolates. Malar J. 2020 Jun 5;19(1):201. doi: 10.1186/s12936-020-03281-x.
• Ref 5: Distribution and Temporal Dynamics of Plasmodium falciparum Chloroquine Resistance Transporter Mutations Associated With Piperaquine Resistance in Northern Cambodia. J Infect Dis. 2021 Sep 17;224(6):1077-1085. doi: 10.1093/infdis/jiab055.
• Ref 6: The malaria parasite's chloroquine resistance transporter is a member of the drug/metabolite transporter superfamily. Mol Biol Evol. 2004 Oct;21(10):1938-49. doi: 10.1093/molbev/msh205. Epub 2004 Jul 7.
• Ref 7: Sequence and gene expression of chloroquine resistance transporter (pfcrt) in the association of in vitro drugs resistance of Plasmodium falciparum. Malar J. 2011 Feb 15;10:42. doi: 10.1186/1475-2875-10-42.
• Ref 8: Glutathione transport: a new role for PfCRT in chloroquine resistance. Antioxid Redox Signal. 2013 Sep 1;19(7):683-95. doi: 10.1089/ars.2012.4625. Epub 2012 Dec 20.
• Ref 9: En-route to the 'elimination' of genotypic chloroquine resistance in Western and Southern Zambia, 14 years after chloroquine withdrawal. Malar J. 2019 Dec 3;18(1):391. doi: 10.1186/s12936-019-3031-4.
• Ref 10: Prevalence of chloroquine and antifolate drug resistance alleles in Plasmodium falciparum clinical isolates from three areas in Ghana. AAS Open Res. 2018 Dec 3;1:1. doi: 10.12688/aasopenres.12825.2. eCollection 2018.
• Ref 11: Structure and drug resistance of the Plasmodium falciparum transporter PfCRT. Nature. 2019 Dec;576(7786):315-320. doi: 10.1038/s41586-019-1795-x. Epub 2019 Nov 27.
• Ref 12: Altered Drug Transport by Plasmodium falciparum Chloroquine Resistance Transporter Isoforms Harboring Mutations Associated with Piperaquine Resistance. Biochemistry. 2020 Jul 14;59(27):2484-2493. doi: 10.1021/acs.biochem.0c00247. Epub 2020 Jul 1.
• Ref 13: Role of Plasmodium falciparum chloroquine resistance transporter and multidrug resistance 1 genes on in vitro chloroquine resistance in isolates of Plasmodium falciparum from Thailand. Am J Trop Med Hyg. 2011 Oct;85(4):606-11. doi: 10.4269/ajtmh.2011.11-0108.
• Ref 14: Survey of Plasmodium falciparum multidrug resistance-1 and chloroquine resistance transporter alleles in Haiti. Malar J. 2013 Nov 19;12:426. doi: 10.1186/1475-2875-12-426.
• Ref 15: Sequence analysis of pfcrt and pfmdr1 genes and its association with chloroquine resistance in Southeast Indian Plasmodium falciparum isolates. Genom Data. 2016 Apr 18;8:85-90. doi: 10.1016/j.gdata.2016.04.010. eCollection 2016 Jun.
• Ref 16: Mutational prevalence of chloroquine resistance transporter gene among Plasmodium falciparum field isolates in Assam and Arunachal Pradesh, India. Indian J Med Microbiol. 2016 Apr-Jun;34(2):193-7. doi: 10.4103/0255-0857.180298.
• Ref 17: Structural and evolutionary analyses of the Plasmodium falciparum chloroquine resistance transporter. Sci Rep. 2020 Mar 16;10(1):4842. doi: 10.1038/s41598-020-61181-1.
• Ref 18: Molecular Mechanisms of Drug Resistance in Plasmodium falciparum Malaria. Annu Rev Microbiol. 2020 Sep 8;74:431-454. doi: 10.1146/annurev-micro-020518-115546.
• Ref 19: The natural function of the malaria parasite's chloroquine resistance transporter. Nat Commun. 2020 Aug 6;11(1):3922. doi: 10.1038/s41467-020-17781-6.
• Ref 20: Phosphomimetic substitution at Ser-33 of the chloroquine resistance transporter PfCRT reconstitutes drug responses in Plasmodium falciparum. J Biol Chem. 2019 Aug 23;294(34):12766-12778. doi: 10.1074/jbc.RA119.009464. Epub 2019 Jul 8.
• Ref 21: Distribution of Mutations Associated with Antifolate and Chloroquine Resistance among Imported Plasmodium vivax in the State of Qatar. Am J Trop Med Hyg. 2017 Dec;97(6):1797-1803. doi: 10.4269/ajtmh.17-0436. Epub 2017 Sep 28.
• Ref 22: Analysis of Plasmodium vivax Chloroquine Resistance Transporter Mutant Isoforms. Biochemistry. 2017 Oct 17;56(41):5615-5622. doi: 10.1021/acs.biochem.7b00749. Epub 2017 Sep 26.
• Ref 23: Verapamil-Sensitive Transport of Quinacrine and Methylene Blue via the Plasmodium falciparum Chloroquine Resistance Transporter Reduces the Parasite's Susceptibility to these Tricyclic Drugs. J Infect Dis. 2016 Mar 1;213(5):800-10. doi: 10.1093/infdis/jiv509. Epub 2015 Oct 26.
• Ref 24: Balancing drug resistance and growth rates via compensatory mutations in the Plasmodium falciparum chloroquine resistance transporter. Mol Microbiol. 2015 Jul;97(2):381-95. doi: 10.1111/mmi.13035. Epub 2015 May 20.
• Ref 25: Expression levels of pvcrt-o and pvmdr-1 are associated with chloroquine resistance and severe Plasmodium vivax malaria in patients of the Brazilian Amazon. PLoS One. 2014 Aug 26;9(8):e105922. doi: 10.1371/journal.pone.0105922. eCollection 2014.
• Ref 26: Drug resistance associated genetic polymorphisms in Plasmodium falciparum and Plasmodium vivax collected in Honduras, Central America. Malar J. 2011 Dec 19;10:376. doi: 10.1186/1475-2875-10-376.
• Ref 27: Characterization of the chloroquine resistance transporter homologue in Toxoplasma gondii. Eukaryot Cell. 2014 Nov;13(11):1360-70. doi: 10.1128/EC.00027-14. Epub 2014 May 23.
• Ref 28: Role of Toxoplasma gondii Chloroquine Resistance Transporter in Bradyzoite Viability and Digestive Vacuole Maintenance. mBio. 2019 Aug 6;10(4):e01324-19. doi: 10.1128/mBio.01324-19.
• Ref 29: Inhibition of efflux of quinolines as new therapeutic strategy in malaria. Curr Top Med Chem. 2008;8(7):563-78. doi: 10.2174/156802608783955593.
• Ref 30: Chloroquine transport via the malaria parasite's chloroquine resistance transporter. Science. 2009 Sep 25;325(5948):1680-2. doi: 10.1126/science.1175667.
• Ref 31: Saquinavir inhibits the malaria parasite's chloroquine resistance transporter. Antimicrob Agents Chemother. 2012 May;56(5):2283-9. doi: 10.1128/AAC.00166-12. Epub 2012 Feb 21.
• Ref 32: Mutations in the Plasmodium falciparum chloroquine resistance transporter, PfCRT, enlarge the parasite's food vacuole and alter drug sensitivities. Sci Rep. 2015 Sep 30;5:14552. doi: 10.1038/srep14552.
• Ref 33: Design, synthesis, and evaluation of 10-N-substituted acridones as novel chemosensitizers in Plasmodium falciparum. Antimicrob Agents Chemother. 2007 Nov;51(11):4133-40. doi: 10.1128/AAC.00669-07. Epub 2007 Sep 10.
• Ref 34: Implications of Glutathione Levels in the Plasmodium berghei Response to Chloroquine and Artemisinin. PLoS One. 2015 May 26;10(5):e0128212. doi: 10.1371/journal.pone.0128212. eCollection 2015.
• Ref 35: A 2-amino quinoline, 5-(3-(2-(7-chloroquinolin-2-yl)ethenyl)phenyl)-8-dimethylcarbamyl-4,6-dithiaoctanoic acid, interacts with PfMDR1 and inhibits its drug transport in Plasmodium falciparum. Mol Biochem Parasitol. 2014 Jun;195(1):34-42. doi: 10.1016/j.molbiopara.2014.05.006. Epub 2014 Jun 8.
• Ref 36: Ex vivo activity of endoperoxide antimalarials, including artemisone and arterolane, against multidrug-resistant Plasmodium falciparum isolates from Cambodia. Antimicrob Agents Chemother. 2014 Oct;58(10):5831-40. doi: 10.1128/AAC.02462-14. Epub 2014 Jul 21.
• Ref 37: Evidence for linkage of pfmdr1, pfcrt, and pfk13 polymorphisms to lumefantrine and mefloquine susceptibilities in a Plasmodium falciparum cross. Int J Parasitol Drugs Drug Resist. 2020 Dec;14:208-217. doi: 10.1016/j.ijpddr.2020.10.009. Epub 2020 Oct 27.
• Ref 38: Piperaquine resistant Cambodian Plasmodium falciparum clinical isolates: in vitro genotypic and phenotypic characterization. Malar J. 2020 Jul 25;19(1):269. doi: 10.1186/s12936-020-03339-w.
• Ref 39: Molecular epidemiology of drug resistance markers of Plasmodium falciparum in Yunnan Province, China. Malar J. 2012 Jul 28;11:243. doi: 10.1186/1475-2875-11-243.
• Ref 40: The Plasmodium falciparum chloroquine resistance transporter is associated with the ex vivo P. falciparum African parasite response to pyronaridine. Parasit Vectors. 2016 Feb 9;9:77. doi: 10.1186/s13071-016-1358-z.
• Ref 41: Polymorphisms in Pfmdr1, Pfcrt, and Pfnhe1 genes are associated with reduced in vitro activities of quinine in Plasmodium falciparum isolates from western Kenya. Antimicrob Agents Chemother. 2014 Jul;58(7):3737-43. doi: 10.1128/AAC.02472-14. Epub 2014 Apr 21.
• Ref 42: Temporal and seasonal changes of genetic polymorphisms associated with altered drug susceptibility to chloroquine, lumefantrine, and quinine in Guinea-Bissau between 2003 and 2012. Antimicrob Agents Chemother. 2015 Feb;59(2):872-9. doi: 10.1128/AAC.03554-14. Epub 2014 Nov 24.
• Ref 43: Ex vivo susceptibilities of Plasmodium vivax isolates from the China-Myanmar border to antimalarial drugs and association with polymorphisms in Pvmdr1 and Pvcrt-o genes. PLoS Negl Trop Dis. 2020 Jun 12;14(6):e0008255. doi: 10.1371/journal.pntd.0008255. eCollection 2020 Jun.
• Ref 44: Distribution of pfmdr1 polymorphisms in Plasmodium falciparum isolated from Southern Thailand. Malar J. 2014 Mar 27;13:117. doi: 10.1186/1475-2875-13-117.
• Ref 45: Phenotypic and genotypic characterization of Thai isolates of Plasmodium falciparum after an artemisinin resistance containment project. Malar J. 2018 May 15;17(1):197. doi: 10.1186/s12936-018-2347-9.
• Ref 46: Mutations in transmembrane domains 1, 4 and 9 of the Plasmodium falciparum chloroquine resistance transporter alter susceptibility to chloroquine, quinine and quinidine. Mol Microbiol. 2007 Jan;63(1):270-82. doi: 10.1111/j.1365-2958.2006.05511.x. Epub 2006 Dec 5.
• Ref 47: Mutation in the Plasmodium falciparum CRT protein determines the stereospecific activity of antimalarial cinchona alkaloids. Antimicrob Agents Chemother. 2012 Oct;56(10):5356-64. doi: 10.1128/AAC.05667-11. Epub 2012 Aug 6.
• Ref 48: Quinine dimers are potent inhibitors of the Plasmodium falciparum chloroquine resistance transporter and are active against quinoline-resistant P. falciparum. ACS Chem Biol. 2014 Mar 21;9(3):722-30. doi: 10.1021/cb4008953. Epub 2014 Jan 6.
• Ref 49: Prevalence of resistance associated polymorphisms in Plasmodium falciparum field isolates from southern Pakistan. Malar J. 2011 Jan 28;10:18. doi: 10.1186/1475-2875-10-18.
• Ref 50: Drug susceptibility of Plasmodium falciparum in eastern Uganda: a longitudinal phenotypic and genotypic study. Lancet Microbe. 2021 Sep;2(9):e441-e449. doi: 10.1016/s2666-5247(21)00085-9. Epub 2021 Jun 18.
• Ref 51: Association of a Novel Mutation in the Plasmodium falciparum Chloroquine Resistance Transporter With Decreased Piperaquine Sensitivity. J Infect Dis. 2017 Aug 15;216(4):468-476. doi: 10.1093/infdis/jix334.
• Ref 52: Globally prevalent PfMDR1 mutations modulate Plasmodium falciparum susceptibility to artemisinin-based combination therapies. Nat Commun. 2016 May 18;7:11553. doi: 10.1038/ncomms11553.