DNA-PK (IC50 <3 nM)

M3814 sensitized multiple tumor cell lines to radiation therapy in vitro [2].

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M3814 targets tumor cell growth and survival by inhibiting a critical DSB DNA damage repair mechanism. The antitumor effect of M3814 is dependent on the functionality of DNA repair and checkpoint signaling in cancer cells, which have a lowered ability to cope with DSBs, leading to cell death. Hence, the rationale of DNA-PK inhibition is to increase the amount of DSB DNA damage generated by IR. The objective of the series of nonclinical experiments was to demonstrate that IR together with M3814 was more efficacious than IR alone [1].

In combination with IR, M3814 showed efficacy in all of the 6 mouse models of human cancer as demonstrated by a strong potentiation of the effect of IR. In all models, a dose of 2 Gy administered daily for 1 week in combination with M3814 induced statically significant tumor growth inhibition compared to IR alone. In 2 models, FaDu and NCIH460, M3814 induced tumor regression. In the FaDu model, no tumor regrowth (duration of the experiment >100 days) was observed in the combination arm with IR (2 Gy per fractions, 6 weeks, 5 days per week: total dose 60 Gy) at the doses of 25 and 50 mg/Kg. In the IR only arm, no tumor responses were observed. These effects were a likely consequence of inhibiting DNA-PK activity, as shown by measuring the autophosphorylation of DNA-PK in FaDu tumor tissue. M3814, alone or in combination with IR, did not induce significant weight loss or visual signs of toxicity in the mice in any study.[1]

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M3814 strongly enhanced the antitumor activity of ionizing radiation in vivo with complete tumor regression applying a clinically relevant fractionated radiation regimen. These effects are due to inhibition of DNA-PK protein kinase activity as demonstrated by the levels of DNA-PK autophosphorylation in human tumor cell lines, and xenograft tumors M3814 is currently investigated in PhI clinical trials [2].

The efficacy of M3814 in combination with IR was evaluated in 6 human xenograft models (HCT116, FaDu, NCI-H460, A549, Capan-1, BxPC3) in mice representing 4 different cancer types (colon, head and neck, lung, and pancreas). Tumor cells were injected s.c. into nude mice, and treatment started when palpable tumors were established (w100-200 mm3 ). M3814 was given orally at different doses (25-300 mg/kg) 10 min prior to IR. IR was applied using a radiation therapy device for small rodents calibrated to deliver 2 Gy. Autophosphorylation of DNA-PK (serine2056 ) in FaDu tumor lysates was measured by immunoassay to assess pharmacological inhibition by M3814.[1]

[1]. M3814, a DNA-dependent Protein Kinase Inhibitor"n(DNA-PKi), Potentiates the Effect of Ionizing Radiation (IR) in"nXenotransplanted Tumors in Nude Mice. IJROBP. 2016; 94, 940-941.

[2]. Abstract 1658: M3814, a novel investigational DNA-PK"ninhibitor: enhancing the effect of fractionated radiotherapy leading to"ncomplete regression of tumors in mice. AACR; Cancer Res 2016;76(14"nSuppl):Abstract nr 1658.

[3]. Arylchinazoline. WO2014183850A1.

Nedisertib is under investigation in clinical trial NCT03770689 (Study of M3814 in Combination With Capecitabine and Radiotherapy in Rectal Cancer).
Peposertib is an orally bioavailable inhibitor of DNA-dependent protein kinase (DNA-PK) with potential antineoplastic activity, and potential sensitizing and enhancing activities for both chemo- and radiotherapies. Upon administration, peposertib binds to and inhibits the activity of DNA-PK, thereby interfering with the non-homologous end joining (NHEJ) process and preventing repair of DNA double strand breaks (DSBs) caused by ionizing radiation or chemotherapeutic treatment. This increases chemo- and radiotherapy cytotoxicity and leads to enhanced tumor cell death. The enhanced ability of tumor cells to repair DSBs plays a major role in the resistance of tumor cells to chemo- and radiotherapy; DNA-PK plays a key role in the NHEJ pathway and DSB repair.

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M3814 is active in nonclinical experiments in combination with IR. Strong antitumor activity was observed in several xenograft models with complete regressions of tumors upon application of the established clinical IR schedule of 2-Gy fractions for 6 weeks in the FaDu model (squamous cell carcinomas of the head and neck). Clinical evaluation of M3814 is ongoing.[1]

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Physical or chemical agents that damage DNA such as ionizing radiation are among the most widely used classes of cancer therapeutics today. Double strand breaks (DSB) generated in DNA by radiation induce multitude of cellular responses, including DNA repair, cell cycle arrest or cell death if the damage is left unrepaired. A complex set of molecular events are responsible for DNA repair via two major mechanisms - homologous recombination (HR) or non-homologous end joining (NHEJ). DNA-PKcs with its regulatory protein subunits, Ku70 and Ku80, is an integral component of NHEJ and considered an attractive intervention point to inhibit DNA repair. We have developed an orally bioavailable, highly potent, and selective inhibitor of DNA-PK, M3814, for cancer therapy in combination with DNA damaging modalities such as radiation, and radio-chemotherapy. Here, we present the preclinical characterization of M3814 using biochemical, cellular and human tumor xenograft models. [2]

C24H21CLFN5O3

481.9066

481.131

C, 59.82; H, 4.39; Cl, 7.36; F, 3.94; N, 14.53; O, 9.96

1637542-33-6

1637542-33-6 (S-isomer); 1637542-34-7 (racemate); 1637542-32-5 (R-isomer)

86292849

Light yellow to yellow solid powder

2.8

1

9

5

34

662

1

ClC1C=C(C(=CC=1[C@@H](C1C=CC(=NN=1)OC)O)C1C2C=CC(=CC=2N=CN=1)N1CCOCC1)F

MOWXJLUYGFNTAL-DEOSSOPVSA-N

InChI=1S/C24H21ClFN5O3/c1-33-22-5-4-20(29-30-22)24(32)16-11-17(19(26)12-18(16)25)23-15-3-2-14(10-21(15)27-13-28-23)31-6-8-34-9-7-31/h2-5,10-13,24,32H,6-9H2,1H3/t24-/m0/s1

(S)-(2-chloro-4-fluoro-5-(7-morpholinoquinazolin-4-yl)phenyl)(6-methoxypyridazin-3-yl)methanol

MSC2490484A; MSC-2490484A; nedisertib; peposertib; 1637542-33-6; M3814; M-3814; MSC-2490484-A; Nedisertib [INN]; M-3814(nedisertib); MSC 2490484A; M3814; M-3814; M 3814

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)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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 (Please use freshly prepared in vivo formulations for optimal results.)