| Size | Price | Stock | Qty |
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| 500mg |
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| 1g |
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| 5g |
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| Other Sizes |
Purity: ≥98%
Chloroquine phosphate is reported to be highly effective in combating SARS-CoV-2 (COVID-19, CoronaVirus, or the COVID-19 pandemic) infection in vitro. It functions as a strong inhibitor of autophagy and toll-like receptors (TLRs) and a 4-aminoquinoline antimalarial drug that is used to treat and prevent malaria in regions where the disease is known to be susceptible to its effects. In addition to being an ATM activator, it also has anti-rheumatoid properties. Chloroquine diphosphate has been documented as an adjuvant for chemotherapy and radiation therapy to induce autophagy in cells and prevent the proliferation or metastasis of anti-cancer cells. The process by which chloroquine diphosphate induces autophagy in cells involves arresting them in the G1 phase, which up-regulates the expression of p53 and p27 and down-regulates the expression of CDK2 and cyclin D1.
| Targets |
Plasmodium; Malaria; TLRs; SARS-COV-2; HIV-1
Chloroquine Phosphate targets p38 mitogen-activated protein kinase (p38 MAPK) [1] Chloroquine Phosphate targets autophagic pathway in cancer cells [2] Chloroquine Phosphate targets 2019-novel coronavirus (2019-nCoV) replication (IC50 = 1.13 μM) [5] Chloroquine Phosphate targets human immunodeficiency virus type 1 (HIV-1) replication [6] |
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| ln Vitro |
In vitro activity: Chloroquine is a chemotherapeutic drug used in the clinical management of malaria. Because chloroquine can bind to DNA, it can prevent RNA synthesis and DNA replication, which ultimately leads to cell death. The creation of a toxic heme-chloroquine complex could potentially be connected to the effects of chloroquine. By raising the pH of vacuoles and blocking the actions of vacuolar phospholipase, vacuolar proteases, and heme polymerase, chloroquine prevents the breakdown of trophozoite hemoglobin. There are specific antirheumatic properties of chloroquine. Due to its immuno-modulatory properties, chloroquine inhibits the release and synthesis of interleukin 6 and tumour necrosis factor. Chloroquine also has direct antiviral effects, preventing the replication of various viruses, including retroviruses, coronaviruses, and flaviviruses, at pH-dependent stages. The effects it has on HIV replication have received the most research. Through the process of ion trapping, chloroquine can accumulate within the macrophage phagolysosome, where it possesses strong antifungal properties against both Cryptococcus neoformans and Histoplasma capsulatum through different mechanisms. While chloroquine directly poisons C. neoformans, it inhibits the growth of H. capsulatum through pH-dependent iron deprivation. In human peripheral blood mononuclear cells (PBMCs) and monocyte-derived Langerhans-like cells (MoLCs) co-culture system, Chloroquine Phosphate (1–10 μM) promotes IL-17 production by CD4+ T cells in a dose-dependent manner. It induces MoLCs to secrete IL-23 (mRNA and protein levels upregulated by ~2.5–3.5 fold), which is dependent on p38 MAPK activation (increased p-p38 expression detected by Western blot). Pretreatment with p38 inhibitor SB203580 reverses the IL-17-promoting effect [1] - In triple-negative breast cancer (TNBC) cell lines (MDA-MB-231, MDA-MB-468), Chloroquine Phosphate exhibits dual effects: low concentrations (0.1–1 μM) promote cell proliferation (viability increased by ~15–25%) and migration (transwell assay shows ~30% increase in migrated cells); high concentrations (10–50 μM) inhibit proliferation (IC50 = 28.5 μM for MDA-MB-231) and induce apoptosis (Annexin V-FITC/PI staining shows apoptotic rate ~40% at 30 μM). It blocks autophagy by increasing LC3-II accumulation and p62 protein levels (Western blot) [2] - In Vero cells infected with 2019-nCoV, Chloroquine Phosphate (0.5–5 μM) inhibits viral replication in a concentration-dependent manner, with an IC50 of 1.13 μM. At 5 μM, it reduces viral load by ~90% compared to control, without significant cytotoxicity (cell viability >80%) [5] - In HIV-1-infected H9 and CEM-T4 cells, Chloroquine Phosphate (5–20 μM) inhibits viral replication: it reduces HIV-1 p24 antigen production by ~50–70% at 10 μM, and blocks viral entry or early post-entry steps (no effect on viral reverse transcriptase activity in cell-free assays) [6] |
| ln Vivo |
Chloroquine (80 mg/kg, i.p.) has no effect on the growth of triple-negative MDA-MB-231 cells with high or low TLR9 expression levels in the orthotopic mouse model. The mouse xenograft model shows significant tumor growth inhibition upon TLR7 and TLR9 inhibition with IRS-954 or chloroquine. Chloroquine also significantly inhibits the development of HCC in the DEN/NMOR rat model. In a mouse model using a subcutaneous xenograft of 4T1 cells, treatment with chloroquine diphosphate dramatically reduced tumor growth and tumor cell metastasis to the lung, improving the mice's survival. Chloroquine diphosphate worked in concert with 5-FU to significantly increase the tumor growth inhibition caused by 5-FU in BALB/c mice that received subcutaneous injections of colon26 cells.This was achieved by increasing the proportion of apoptotic cells.
In a subcutaneous xenograft model of TNBC (MDA-MB-231 cells implanted in nude mice), intraperitoneal administration of Chloroquine Phosphate shows dual effects: low dose (5 mg/kg/day) promotes tumor growth (tumor volume increased by ~30% vs. control); high dose (25 mg/kg/day) inhibits tumor growth by ~55% vs. control. High-dose treatment increases apoptotic cells (TUNEL assay) and LC3-II/p62 accumulation in tumor tissues, confirming autophagy inhibition [2] |
| Enzyme Assay |
Chloroquine suppressed matrix metalloproteinase (MMP)-2 and MMP-9 mRNA expression and protein activity, whereas MMP-13 mRNA expression and proteolytic activity were increased. Despite enhancing TLR9 mRNA expression, chloroquine suppressed TLR9 protein expression in vitro.[2]
p38 MAPK activity assay: MoLCs were treated with Chloroquine Phosphate (1–10 μM) for 24 hours, and cell lysates were prepared. Recombinant p38 substrate peptide and ATP were added to the lysates, and the mixture was incubated at 30°C for 60 minutes. Phosphorylated peptide was detected by ELISA, and p38 activation was quantified by the ratio of phosphorylated to total peptide. The assay confirmed that Chloroquine Phosphate activates p38 MAPK in MoLCs [1] - HIV-1 reverse transcriptase (RT) activity assay: Cell-free HIV-1 RT was incubated with poly(rA)-oligo(dT) substrate, [3H]-TTP, and Chloroquine Phosphate (5–50 μM) at 37°C for 90 minutes. Radioactivity of the synthesized DNA was measured by liquid scintillation counting. No significant inhibition of RT activity was observed, indicating Chloroquine Phosphate acts on viral entry/post-entry steps rather than RT [6] |
| Cell Assay |
The cells are cultivated in 6-well plates using normal culture medium with either vehicle or 25 or 50 μM chloroquine until they are almost confluent. Afterward, they are rinsed with sterile phosphate-buffered saline (PBS) and continue to culture in serum-free culture medium for the specified durations. Centrifugation is used to quickly harvest the cells in lysis buffer and clarify them after the culture medium is discarded at the predetermined time points. Once the supernatants have been boiled in reducing sodium dodecyl sulphate (SDS) sample buffer, 100 μg of protein is loaded into each lane. The samples are then electrophoresed into 10 or 4–20% gradient polyacrylamide SDS gels before being placed onto a nitrocellulose membrane. TLR9 is detected by incubating the blots with anti-TLR9 antibodies diluted 1:500 in Tris-buffered saline containing 0.1% (v/v) Tween-20 (TBST) for an overnight period at 4°C. Polyclonal rabbit anti-actin is used to confirm equal loading. Horseradish peroxidase-linked secondary antibodies are used in secondary detection. Using an ECL kit, the protein bands can be seen through chemiluminescence.
CD4+ T cell IL-17 production assay: PBMCs were isolated and MoLCs were differentiated from monocytes. MoLCs were pretreated with Chloroquine Phosphate (1–10 μM) for 24 hours, then co-cultured with CD4+ T cells (isolated by magnetic sorting) for 72 hours. IL-17 levels in the supernatant were measured by ELISA, and CD4+IL-17+ T cells were quantified by flow cytometry [1] - TNBC cell proliferation and apoptosis assay: MDA-MB-231/MDA-MB-468 cells were seeded in 96-well plates, treated with Chloroquine Phosphate (0.1–50 μM) for 48 hours. Cell viability was detected by CCK-8 assay; apoptosis was analyzed by Annexin V-FITC/PI staining and flow cytometry. For autophagy detection, cells were transfected with GFP-LC3 plasmid, treated with the drug, and LC3 puncta were observed by fluorescence microscopy [2] - 2019-nCoV inhibition assay: Vero cells were seeded in 96-well plates and infected with 2019-nCoV (MOI = 0.01) for 1 hour. Chloroquine Phosphate (0.5–5 μM) was added, and cells were cultured for 48 hours. Viral RNA was extracted, and viral load was quantified by qRT-PCR (ORF1ab gene); cell viability was measured by CCK-8 assay to exclude cytotoxicity [5] - HIV-1 inhibition assay: H9/CEM-T4 cells were infected with HIV-1 (strain IIIB) at MOI = 0.01, then treated with Chloroquine Phosphate (5–20 μM) for 72 hours. HIV-1 p24 antigen levels in the supernatant were measured by ELISA, and infected cells were detected by flow cytometry using anti-p24 antibody [6] |
| Animal Protocol |
Mice with impaired immunity, four weeks old, have their mammary fat pads injected with a combination of control and TLR9 siRNA MDA-MB-231 cells (5×105 cells in 100 μL) (athymic nude/nu Foxn1). Seven days after the tumor cell inoculation, treatments begin. Every day, the mice receive either a vehicle (PBS) or intraperitoneal (i.p.) chloroquine (80 mg/kg). Every day, the animals are observed for clinical indications. Tumor volumes are estimated using the formula V=(π/6) (d1×d2)3/2, where d1 and d2 are the perpendicular tumor diameters. Tumor measurements are carried out twice a week. The mice are killed and the tumors are removed for one last measurement after the tumors are left to grow for 22 days. The animals are kept in controlled, pathogen-free environments with 20–21°C temperatures, 30–60% relative humidity, and a 12-hour lighting cycle throughout the trials. Small-animal food pellets are fed to the mice, and they are given access to sterile water whenever they need it.
TNBC xenograft model: Nude mice (4-week-old, female) were subcutaneously injected with MDA-MB-231 cells (5×106 cells/mouse) into the right flank. When tumors reached ~100 mm3, mice were randomly divided into control, low-dose (5 mg/kg/day), and high-dose (25 mg/kg/day) Chloroquine Phosphate groups (n = 8 per group). The drug was dissolved in normal saline and administered via intraperitoneal injection once daily for 21 days. Tumor volume was measured every 3 days (volume = length × width2 / 2), and mice were euthanized for tumor weight measurement and immunohistochemical analysis (LC3, p62, Ki-67, TUNEL) [2] |
| Toxicity/Toxicokinetics |
Effects During Pregnancy and Lactation
◉ Overview of Medication Use During Lactation Very small amounts of chloroquine are excreted into breast milk; a weekly dose is insufficient to harm the infant or protect them from malaria infection. The UK Malaria Treatment Guidelines recommend 500 mg of chloroquine weekly until the end of lactation and when primaquine can be administered. Breastfeeding infants should receive the recommended dose of chloroquine for malaria prevention. In HIV-infected women, women treated with chloroquine showed a more significant decrease in HIV viral load in breast milk compared to women receiving a combination of sulfadoxine and pyrimethamine. Since there is currently no information on daily chloroquine use during lactation, hydroxychloroquine or other medications may be a better option in such cases, especially for breastfeeding newborns or premature infants. ◉ Impact on Breastfed Infants Several authors have noted that in malaria-endemic areas, it is common for breastfeeding mothers to take chloroquine for malaria prevention. As of the revision date, no reports of adverse reactions in breastfed infants have been published. ◉ Effects on lactation and breast milk As of the revision date, no relevant published information was found. In vitro cytotoxicity: Chloroquine phosphate at concentrations up to 5 μM showed no significant cytotoxicity to Vero cells, H9/CEM-T4 cells, or normal human mammary epithelial cells (HMEC) (cell viability >80% vs. control group) [5,6,2] - In vivo toxicity: Nude mice treated with chloroquine phosphate (25 mg/kg/day, intraperitoneal injection, for 21 days) did not show significant weight loss, lethargy, or organ damage. Serum biochemical analysis (ALT, AST, BUN, creatinine) and histological examination of liver, kidney and heart tissues showed no abnormalities [2] - Plasma protein binding rate: The plasma protein binding rate of chloroquine phosphate in human plasma is approximately 55-65% (the study cited in the relevant literature review used the balanced dialysis method to determine this) [4] |
| References | |
| Additional Infomation |
Chloroquine phosphate is the phosphate salt of chloroquine, a quinoline compound with antimalarial and anti-inflammatory properties. Chloroquine is currently the most widely used antimalarial drug, except for malaria caused by chloroquine-resistant Plasmodium falciparum. Although its mechanism of action is not fully understood, studies have shown that chloroquine inhibits heme polymerase, an enzyme in parasites that converts toxic heme into non-toxic heme, leading to the accumulation of toxic heme within the parasite. Chloroquine may also interfere with nucleic acid biosynthesis. See also: chloroquine (containing the active moiety); chloroquine phosphate; primaquine phosphate (component); chloroquine phosphate; embutrothion; lidocaine (component).
Chloroquine phosphate is a synthetic antimalarial drug that, in addition to its antimalarial effects, has a variety of pharmacological effects, including immunomodulation, autophagy inhibition and antiviral activity[4] - Its immunomodulatory mechanism involves activating p38 MAPK in monocyte lymphocytes (MoLCs), promoting IL-23 secretion, and then inducing CD4+ T cells to produce IL-17, which may regulate the inflammatory response[1] - In triple-negative breast cancer (TNBC), its dual role is associated with autophagy regulation: low doses mildly inhibit autophagy to promote cell survival, while high doses strongly block autophagy, leading to apoptosis[2] - As an antiviral drug, it inhibits viral replication by altering endosomal pH (blocking viral fusion/entry) and interfering with viral protein processing, and is active against 2019-nCoV, HIV-1 and other enveloped viruses. [5,6,4] - This drug has been used clinically to treat malaria and rheumatoid arthritis, and has been repurposed for the treatment of COVID-19 in preclinical and early clinical studies. [4] |
| Molecular Formula |
C18H32CLN3O8P2
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| Molecular Weight |
515.8625
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| Exact Mass |
515.14
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| Elemental Analysis |
C, 41.91; H, 6.25; Cl, 6.87; N, 8.15; O, 24.81; P, 12.01
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| CAS # |
50-63-5
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| Related CAS # |
Chloroquine phosphate;50-63-5;Chloroquine-d5;1854126-41-2;Chloroquine dihydrochloride;3545-67-3;Chloroquine-d5 diphosphate; 132-73-0 (sulfate); 1854126-42-3; 54-05-7 ;151-69-9 (acetate) ; 1446-17-9 (phosphate); 3545-67-3 (HCl) ; 50-63-5 (diphosphate) ;
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| PubChem CID |
64927
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| Appearance |
White to light yellow crystalline powder
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| Boiling Point |
460.6ºC at 760 mmHg
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| Melting Point |
200 °C
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| Flash Point |
232.3ºC
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| LogP |
3.03
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| Hydrogen Bond Donor Count |
7
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| Hydrogen Bond Acceptor Count |
11
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| Rotatable Bond Count |
8
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| Heavy Atom Count |
32
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| Complexity |
359
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
QKICWELGRMTQCR-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C18H26ClN3.2H3O4P/c1-4-22(5-2)12-6-7-14(3)21-17-10-11-20-18-13-15(19)8-9-16(17)18;2*1-5(2,3)4/h8-11,13-14H,4-7,12H2,1-3H3,(H,20,21);2*(H3,1,2,3,4)
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| Chemical Name |
4-N-(7-chloroquinolin-4-yl)-1-N,1-N-diethylpentane-1,4-diamine;phosphoric acid
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| Synonyms |
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
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| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: 100 mg/mL (193.85 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication (<60°C).
(Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.9385 mL | 9.6926 mL | 19.3851 mL | |
| 5 mM | 0.3877 mL | 1.9385 mL | 3.8770 mL | |
| 10 mM | 0.1939 mL | 0.9693 mL | 1.9385 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
| NCT Number | Recruitment | interventions | Conditions | Sponsor/Collaborators | Start Date | Phases |
| NCT03529396 | Active Recruiting |
Drug: Chloroquine Drug: Primaquine |
Vivax Malaria G6PD Deficiency |
Fundação de Medicina Tropical Dr. Heitor Vieira Dourado |
July 20, 2018 | Phase 2 |
| NCT02932007 | Active Recruiting |
Drug: Chloroquine Phosphate | Atrial Fibrillation | University of South Florida | March 28, 2017 | Phase 2 |
| NCT06076837 | Not yet recruiting | Drug: Chloroquine Phosphate Drug: Balstilimab |
Pancreatic Cancer Metastatic | HonorHealth Research Institute | December 2023 | Phase 1 |
| NCT05443178 | Recruiting | Drug: Nivaquine ® (Chloroquine) | Tuberculosis Infection | University of Zurich | January 4, 2022 | Phase 1 |
| NCT04704999 | Not yet recruiting | Drug: Chloroquine Drug: Tafenoquine |
Plasmodium Vivax Malaria | University of Oxford | September 18, 2023 | Phase 4 |
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