VEGFR2 (IC50 = 1 nM); RET (IC50 = 13 nM)

Apatinib significantly and dose-dependently inhibits the growth of a variety of human tumor xenografts. The ABCB1-mediated MDR in the nude mouse xenograft model is reversed by aparinib. Apatinib dramatically increases doxorubicin's antitumor activity in nude mice that have K562/ADR xenografts [3]

Apatinib significantly and dose-dependently inhibits the growth of a variety of human tumor xenografts. The ABCB1-mediated MDR in the nude mouse xenograft model is reversed by aparinib. Apatinib dramatically increases doxorubicin's antitumor activity in nude mice that have K562/ADR xenografts.[3]

The substrate solution containing tyrosine is a poly(glu, ala, tyr) 6:3:1 random copolymer. In order to coat 96 well plates (100 L/well), the substrate is kept as a 1 mg/mL stock in PBS at 20 °C and diluted 1 in 500 with PBS. The day before the assay, plates are coated, sealed with adhesive seals, and kept at 4 °C for the entire night. The substrate solution is disposed of on the day of the assay, and the assay plate wells are twice washed—once with Hepes buffer (50 mM, pH 7.4) and once with PBST (PBS containing 0.05% v/v Tween 20). The assay plates are washed and the test compounds are diluted with 10% dimethylsulfoxide (DMSO) de-ionized water, with 25 μL volumes being transferred to the wells. All test wells are then filled with 25 μL of a manganese chloride solution (40 mM) containing 8 μM ATP. To find the assay's dynamic range, additional wells are added that contain blank and control solutions, respectively, containing manganese chloride solution with and without ATP and compound diluent. Each well receives 50 L of freshly diluted enzyme, which is then added. The plates are then left to sit at room temperature for 20 min. Subsequently, the liquid is disposed of and the wells are twice cleaned using PBST. After adding 100 L/well of mouse IgG anti-phosphotyrosine antibody diluted 1:6000 with PBST containing 0.5% (w/v) bovine serum albumin (BSA), the plates were incubated for 1h at room temperature before the liquid was discarded and the wells were twice washed with PBST. The sheep anti-mouse Ig antibody linked to horseradish peroxidase (HRP) is diluted 1:500 with PBST containing 0.5% (w/v) BSA. After adding 100 μL/well, the plates are incubated for an additional hour at room temperature. The liquid is then discarded, and the wells are twice washed with PBST. Freshly prepared 50 mM phosphate-citrate buffer (pH 5.0) containing 0.03% (w/v) sodium perborate is mixed with 1 mg/mL of 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid, and 100 μL is added to each well. The plates are then incubated at room temperature for 20 to 60 minutes, or until the control wells' optical density value, measured at 405 nm, is roughly 1.0. Microcal Origin is used to interpolate IC50 values for compound enzyme inhibition after subtracting blank values.

In 96-well plates, the HUVEC are seeded. The test agents (vehicle serving as the control) are added to the cells along with 20 ng/mL VEGF or 20% FBS and incubated for an additional 72 hours. Following 10% trichloroacetic acid fixation, the cells are stained for 30 minutes at 37 °C using 0.4% sulforhodamine B, and then they are cleaned with 1% acetic acid. After dissolving the complex with tris, the optical density is measured at 520 nM.

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YN968D1 both by itself and in conjunction with chemotherapeutic agents efficiently and minimally harmed the growth of multiple well-established human tumor xenograft models in vivo[1].

Ls174t, HCT 116, SGC-7901, HT-29, A549, NCI-H460 xenografted BALB/cA nude mice
50, 100, 200 mg/kg
p.o.

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Nude mouse human tumor xenograft model.  The effects of Apatinib (YN968D1) on tumor growth were tested against various human tumors grown subcutaneously in BALB/cA nude mice. Tumor growth was initiated by subcutaneous inoculation of cells into mice. Tumors were allowed to establish and grow to 100–300 mm3, at which time the mice were randomized into experimental groups. YN968D1 was administered once daily by oral gavage for the indicated periods (Table 1). In combination treatment experiments, mice were administered YN968D1 alone by oral gavage; 5‐FU, oxaliplatin, docetaxel and doxorubicin alone by intravenous injection; or YN968D1 in combination with each cytotoxic drug at the indicated dose and schedule (Table 2). Tumor volume and bodyweight were monitored every other day or every 3 days, with the means indicated for groups of six (treated) or 12 (vehicle control) animals. Tumor volumes were determined by measuring the largest diameter (a) and its perpendicular (b) according to the formula (a × b2)/2. The evaluation index for inhibition was the relative tumor growth ratio according to the equation: T/C (%) = mean increase of tumor volumes of treated groups/mean increase of tumor volumes of control groups × 100%.[1]

[1]. Cancer Sci . 2011 Jul;102(7):1374-80.

[2]. Cancer Res . 2010 Oct 15;70(20):7981-91.

[3]. Biochem Pharmacol . 2012 Mar 1;83(5):586-97.

Rivoceranib Mesylate is the mesylate salt of rivoceranib, an orally bioavailable, small-molecule receptor tyrosine kinase inhibitor with potential antiangiogenic and antineoplastic activities. Rivoceranib selectively binds to and inhibits vascular endothelial growth factor receptor 2, which may inhibit VEGF-stimulated endothelial cell migration and proliferation and decrease tumor microvessel density. In addition, this agent mildly inhibits c-Kit and c-SRC tyrosine kinases.

C25H27N5O4S

493.58

493.17837553

C, 60.83; H, 5.51; N, 14.19; O, 12.97; S, 6.50

1218779-75-9

1218779-89-5 (HCl);1218779-75-9 (mesylate);811803-05-1;

45139106

white solid powder

1.3±0.1 g/cm3

578.2±50.0 °C at 760 mmHg

303.5±30.1 °C

0.0±1.6 mmHg at 25°C

1.652

3.68

3

8

6

35

701

0

S(C([H])([H])[H])(=O)(=O)O[H].O=C(C1C([H])=C([H])C([H])=NC=1N([H])C([H])([H])C1C([H])=C([H])N=C([H])C=1[H])N([H])C1C([H])=C([H])C(=C([H])C=1[H])C1(C#N)C([H])([H])C([H])([H])C([H])([H])C1([H])[H]

FYJROXRIVQPKRY-UHFFFAOYSA-N

InChI=1S/C24H23N5O.CH4O3S/c25-17-24(11-1-2-12-24)19-5-7-20(8-6-19)29-23(30)21-4-3-13-27-22(21)28-16-18-9-14-26-15-10-18;1-5(2,3)4/h3-10,13-15H,1-2,11-12,16H2,(H,27,28)(H,29,30);1H3,(H,2,3,4)

N-[4-(1-cyanocyclopentyl)phenyl]-2-(pyridin-4-ylmethylamino)pyridine-3-carboxamide;methanesulfonic acid

YN968D1 mesylate; YN-968D1 mesylate; YN 968D1 mesylate; Rivoceranib mesylate

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO: ~22 mg/mL (~44.6 mM)
Water: > 10mg/mL
Ethanol: < 1 mg/mL