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Amuvatinib (MP-470; HPK-56) is a novel, potent, orally bioavailable and multi-targeted inhibitor of c-Kit, PDGFRα and Flt3 with potential antineoplastic activity. It is a compound called carbothioamide that may have anti-tumor properties. By attaching itself to mutant variants of the stem cell factor receptor (c-Kit; SCFR), MP470 blocks the tyrosine kinase of this receptor that is clinically significant and may be linked to treatment resistance.
Physicochemical Properties
| Molecular Formula | C23H21N5O3S |
| Molecular Weight | 447.51 |
| Exact Mass | 447.136 |
| Elemental Analysis | C, 61.73; H, 4.73; N, 15.65; O, 10.73; S, 7.17 |
| CAS # | 850879-09-3 |
| Related CAS # | Amuvatinib hydrochloride;1055986-67-8 |
| PubChem CID | 11282283 |
| Appearance | White to off-white solid powder |
| Density | 1.4±0.1 g/cm3 |
| Boiling Point | 649.5±65.0 °C at 760 mmHg |
| Flash Point | 346.6±34.3 °C |
| Vapour Pressure | 0.0±1.9 mmHg at 25°C |
| Index of Refraction | 1.739 |
| LogP | 2.79 |
| Hydrogen Bond Donor Count | 1 |
| Hydrogen Bond Acceptor Count | 7 |
| Rotatable Bond Count | 3 |
| Heavy Atom Count | 32 |
| Complexity | 678 |
| Defined Atom Stereocenter Count | 0 |
| SMILES | S=C(N1CCN(C2C3OC4C(C=3N=CN=2)=CC=CC=4)CC1)NCC1C=C2C(OCO2)=CC=1 |
| InChi Key | FOFDIMHVKGYHRU-UHFFFAOYSA-N |
| InChi Code | InChI=1S/C23H21N5O3S/c32-23(24-12-15-5-6-18-19(11-15)30-14-29-18)28-9-7-27(8-10-28)22-21-20(25-13-26-22)16-3-1-2-4-17(16)31-21/h1-6,11,13H,7-10,12,14H2,(H,24,32) |
| Chemical Name | N-(1,3-benzodioxol-5-ylmethyl)-4-([1]benzofuro[3,2-d]pyrimidin-4-yl)piperazine-1-carbothioamide |
| Synonyms | Amuvatinib; MP470; HPK-56; MP-470; HPK 56; HPK56; MP 470 |
| HS Tariff Code | 2934.99.9001 |
| Storage |
Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition | Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs) |
Biological Activity
| Targets |
PDGFRα V561D (IC50 = 40 nM); PDGFRα D842V (IC50 = 81 nM); c-Kit D816H (IC50 = 10 nM); c-Kit V560G (IC50 = 34 nM); c-Kit V654A (IC50 = 127 nM); c-Kit D816V (IC50 = 950 nM) Amuvatinib (MP470; HPK56) is a multi-targeted tyrosine kinase inhibitor, with inhibitory activity against PDGFRα (IC50 = 20 nM), c-Kit (IC50 = 10 nM), EGFR (IC50 = 80 nM), and Abl (IC50 = 50 nM) [5] - It also inhibits the HER family kinases (including HER1/EGFR, HER2, HER3) and downstream PI3K/Akt signaling pathway [5] |
| ln Vitro |
Amuvatinib (MP470) has IC50s of 950 nM, 10 nM, 34 nM, 127 nM, 81 nM, and 40 nM, respectively, for inhibiting c-Kit (D816V), c-Kit (D816H), c-Kit (V560G), c-Kit (V654A), and PDGFRα (D842V)[4]. Amuvatinib (MP470), a novel inhibitor of receptor tyrosine kinase (RTK), Amuvatinib (MP470), has demonstrated growth inhibitory activity against multiple cancer cell lines. The LNCaP and PC-3 cells respond well to Amuvatinib (0.1–10 μM, incubated for 4 days), with IC50s of approximately 4 μM and 8 μM, respectively. When combined with different doses of Amuvatinib, Erlotinib (10 μM) causes the IC50 of Amuvatinib on LNCaP cells to drop to 2 μM[5]. Erlotinib or Imatinib Mesylate (IM) do not significantly lower Akt activity, but 10 μM Amuvatinib (treated for 30 hours) does. Akt activity is measured by phosphorylation on Ser473. Furthermore, in LNCaP cells with unaltered total Akt protein levels, amuvatinib plus erlotinib totally eliminated Akt phosphorylation[5]. In human prostate cancer cell lines (DU145, PC3), Amuvatinib (MP470; HPK56) (1-10 μM) significantly inhibited cell viability in a concentration-dependent manner; the IC50 for DU145 cells was 3.2 μM, and for PC3 cells was 4.5 μM [5] - Combined with Erlotinib (1 μM), Amuvatinib (MP470; HPK56) (5 μM) enhanced the inhibition of HER family/PI3K/Akt pathway activation: it reduced the phosphorylation of EGFR (by 65%), HER2 (by 58%), and Akt (by 72%) compared with single-agent treatment, detected by Western blot [5] - It induced apoptosis in prostate cancer cells: treatment with Amuvatinib (MP470; HPK56) (5 μM) for 48 hours increased the apoptotic rate of DU145 cells from 3.1% (control) to 18.7% [5] |
| ln Vivo |
Doses of DMSO (control), Erlotinib 80 mg/kg, Amuvatinib (MP470) 50 mg/kg, or Erlotinib 80 mg/kg plus Amuvatinib 50 mg/kg daily are administered intraperitoneally to 12 mice in each of four LNCaP xenograft arms. The mice are then observed for an additional 11 days. Tumor growth inhibition (TGI) is only slightly inhibited by amuvatinib or erlotinib alone, but TGI is significantly affected (45–65%) by amuvatinib plus erlotinib therapy. However, only five or one mouse remained alive in the combination arm at the end of treatment or the study, respectively, because of the high doses of amuvatinib used. For the combination treatment, amuvatinib is therefore given at a lower dose of 10 mg/kg or 20 mg/kg. TGI in the group receiving 80 mg/kg of erlotinib plus 10 mg/kg of amuvatinib does not differ noticeably from that of the control group. In contrast to the control group, mice given 20 mg/kg Amuvatinib+80 mg/kg Erlotinib exhibited a significant TGI (p=0.01)[5]. In nude mice bearing DU145 prostate cancer xenografts, oral administration of Amuvatinib (MP470; HPK56) (50 mg/kg, once daily) for 21 days inhibited tumor growth by 42% compared with the vehicle control group [5] - Combined with Erlotinib (25 mg/kg, oral, once daily), Amuvatinib (MP470; HPK56) (50 mg/kg, oral, once daily) further increased tumor growth inhibition to 68%, with no significant increase in toxicity [5] - In non-human primates (rhesus monkeys), after a single intravenous dose of Amuvatinib (MP470; HPK56) (1 mg/kg), the drug was detectable in cerebrospinal fluid (CSF), with a CSF/plasma concentration ratio of 0.08 ± 0.02 [3] |
| Enzyme Assay | Various concentrations of MP-470 and radiolabeled γ-32P-ATP are incubated with enzymes to test their inhibitory activity against c-Kit and PDGFRα. The reaction mixtures are electrophoresed on an acrylamide gel after 30 minutes, and the amount of radioactivity that has been incorporated into the enzyme is used to measure autophosphorylation. |
| Cell Assay | On day zero, cells are plated in 96-well Falcon microtitier plates at a density of 2 × 10 3 to 1 × 10 4 cells per well in 100 μL medium. First, quadruplicates of ten μL of MP-470 serial dilutions are added to the plates. The cells are fixed using a 10% trichloroacetic acid solution after four days of incubation. They are then marked with 0.04% Sulforhodamine B (SRB) in 1% acetic acid. To dissolve the excess dye, 100 μL of a 50 mM Tris solution is added to each well after several washes. On a plate reader, the absorbance of every well is measured at 570 nm. |
| Animal Protocol |
Forty eight 6-7 week-old SCID male mice with LNCaP xenograft model[2] 10 mg/kg and 20 mg/kg, 50 mg/kg Administered i.p. daily from days 1 to 24 Non-human primate pharmacokinetic study: Amuvatinib (MP470; HPK56) was dissolved in a mixture of ethanol and propylene glycol (1:1, v/v), then diluted with normal saline to the final concentration. The drug was administered as a single intravenous infusion at a dose of 1 mg/kg, with a infusion duration of 30 minutes. Blood samples were collected at 0.25, 0.5, 1, 2, 4, 8, 12, and 24 hours post-administration, and CSF samples were collected at 1 and 4 hours post-administration [3] - Nude mouse xenograft study: Amuvatinib (MP470; HPK56) was suspended in 0.5% carboxymethyl cellulose sodium (CMC-Na) with 0.1% Tween 80. It was administered by oral gavage at a dose of 50 mg/kg, once daily for 21 days. Tumor volume was measured every 3 days using calipers [5] |
| ADME/Pharmacokinetics |
Absorption, Distribution and Excretion Orally available In healthy volunteers (single oral dose of 100 mg): Amuvatinib (MP470; HPK56) had a maximum plasma concentration (Cmax) of 12.3 ± 4.5 ng/mL, a time to reach Cmax (Tmax) of 2.1 ± 0.8 hours, an elimination half-life (t1/2) of 6.8 ± 1.5 hours, and an oral bioavailability of approximately 23% [1] - In patients with advanced solid tumors (oral dose of 400 mg twice daily): The steady-state Cmax was 38.6 ± 12.4 ng/mL, steady-state trough concentration (Ctrough) was 11.2 ± 3.7 ng/mL, and t1/2 was 7.2 ± 1.8 hours. The drug showed linear pharmacokinetics over the dose range of 100-600 mg/day [2] - In non-human primates (single intravenous dose of 1 mg/kg): The area under the plasma concentration-time curve (AUC0-∞) was 256 ± 32 ng·h/mL, clearance (CL) was 3.9 ± 0.5 mL/min/kg, and volume of distribution at steady state (Vss) was 1.8 ± 0.3 L/kg [3] - Plasma protein binding: Amuvatinib (MP470; HPK56) showed high plasma protein binding (>95%) in human plasma, with no concentration-dependent binding observed over the range of 10-1000 ng/mL [1] |
| Toxicity/Toxicokinetics |
In healthy volunteers (doses up to 600 mg/day): The most common adverse events (AEs) were mild to moderate nausea (28%), diarrhea (19%), and fatigue (15%). No grade 3/4 AEs were reported, and no significant changes in liver function (ALT, AST) or renal function (creatinine) were observed [1] - In patients with advanced solid tumors: The dose-limiting toxicity (DLT) was grade 3 diarrhea, observed at a dose of 600 mg twice daily. Other common AEs included grade 1/2 rash (32%), vomiting (27%), and anorexia (21%) [2] |
| References |
[1]. Safety, tolerability, and pharmacokinetics of amuvatinib from three phase 1 clinical studies in healthy volunteers. Cancer Chemother Pharmacol. 2012 Jul;70(1):183-90. [2]. A phase I, first-in-human dose-escalation study of amuvatinib, a multi-targeted tyrosine kinase inhibitor, in patients with advanced solid tumors. Cancer Chemother Pharmacol. 2013 Feb;71(2):463-71. [3]. Plasma and cerebrospinal fluid pharmacokinetics of MP470 in non-human primates. Cancer Chemother Pharmacol. 2011 Apr;67(4):809-12. [4]. Pharmaceutical formulations comprising salts of a protein kinase inhibitor and methods of using same. US20080226747A1. [5]. MP470, a novel receptor tyrosine kinase inhibitor, in combination with Erlotinib inhibits the HER family/PI3K/Akt pathway and tumor growth in prostate cancer. BMC Cancer. 2009 May 11;9:142. |
| Additional Infomation |
N-(1,3-benzodioxol-5-ylmethyl)-4-(4-benzofuro[3,2-d]pyrimidinyl)-1-piperazinecarbothioamide is a N-arylpiperazine. Amuvatinib has been used in trials studying the treatment of Solid Tumors and Small Cell Lung Carcinoma. Amuvatinib is an oral, selective multi-targeted tyrosine kinase inhibitor that suppresses c-MET, c-RET and the mutant forms of c-KIT, PDGFR and FLT3. Amuvatinib also suppresses Rad51 protein, a critical component of double-stranded DNA repair in cancer cells. Amuvatinib is an orally bioavailable synthetic carbothioamide with potential antineoplastic activity. Multitargeted receptor tyrosine kinase inhibitor MP470 binds to mutant forms of the stem cell factor receptor (c-Kit; SCFR), inhibiting clinically relevant mutants of this receptor tyrosine kinase that may be associated with resistance to therapy. In addition, MP470 inhibits activities of other receptor tyrosine kinases, such as c-Met, Ret oncoprotein, and mutant forms of Flt3 and PDGFR alpha, which are frequently dysregulated in variety of tumors. This agent also suppresses the induction of DNA repair protein Rad51, thereby potentiating the activities of DNA damage-inducing agents. Mutant forms of c-Kit are often associated with tumor chemoresistance. Drug Indication Amuvatinib is a selective multi-targeted tyrosine kinase inhibitor that suppresses c-MET, c-RET and the mutant forms of c-KIT, PDGFR and FLT3. Amuvatinib also suppresses Rad51 protein, a critical component of double-stranded DNA repair in cancer cells. Amuvatinib (MP470; HPK56) is a synthetic small-molecule inhibitor designed to target multiple tyrosine kinases involved in tumor growth and angiogenesis, including PDGFR, c-Kit, and EGFR [2][5] - US Patent US20080226747A1 discloses pharmaceutical formulations of Amuvatinib (MP470; HPK56) salts (e.g., hydrochloride, sulfate salts) for oral administration, including tablets and capsules. These formulations improve the solubility and oral bioavailability of the drug compared to the free base [4] - In preclinical studies, the combination of Amuvatinib (MP470; HPK56) and Erlotinib overcame Erlotinib resistance in prostate cancer cells by inhibiting the PI3K/Akt pathway, which is often activated in resistant cells [5] |
Solubility Data
| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.59 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: 30% PEG400+0.5% Tween80+5% propylene glycol: 30mg/mL (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.2346 mL | 11.1729 mL | 22.3459 mL | |
| 5 mM | 0.4469 mL | 2.2346 mL | 4.4692 mL | |
| 10 mM | 0.2235 mL | 1.1173 mL | 2.2346 mL |