CN115124528B - Pyrrolopyridine compound, and preparation method and medical application thereof - Google Patents

Pyrrolopyridine compound, and preparation method and medical application thereof Download PDF

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CN115124528B
CN115124528B CN202210850237.3A CN202210850237A CN115124528B CN 115124528 B CN115124528 B CN 115124528B CN 202210850237 A CN202210850237 A CN 202210850237A CN 115124528 B CN115124528 B CN 115124528B
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CN115124528A (en
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牟杰
周婷
徐雯
郭俊宇
强国威
凌心迪
裴冬生
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Xuzhou Medical University
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Abstract

The application relates to a pyrrolopyridine compound, a preparation method and medical application thereof, belonging to the technical fields of pharmaceutical chemistry and pharmacotherapeutics. The compound shown in the formula I, isomer, hydrate, solvate or pharmaceutically acceptable salt thereof has good PAK5 inhibition activity, can be used for preparing medicines related to serine/threonine kinase PAK5, and has stronger anti-tumor activity on tumor cells such as kidney cancer cells, liver cancer cells, colorectal cancer cells, breast cancer cells and the like.

Description

Pyrrolopyridine compound, and preparation method and medical application thereof
Technical Field
The application belongs to the technical fields of pharmaceutical chemistry and pharmacotherapeutics, and particularly relates to a pyrrolopyridine compound. The compounds can be used for preparing medicines for treating diseases related to PAK 5. The application also relates to a preparation method of the compounds and a pharmaceutical composition containing the compounds.
Background
The pyrrole [2,3-b ] pyridine is an important nitrogenous heterocyclic compound, has similar structures as indole, purine and the like, becomes a bioisostere of the latter two compounds, and has wide application in the fields of medicines, pesticides and the like. The natural product contains pyrrolopyridine structure compound which is an important alkaloid and has physiological activities of resisting cancer, bacteria, psychosis and the like. The pyrrolopyridine derivative has the function of inhibiting protein kinase, and has potential biological activity and medicinal value in aspects of antihistamine, anti-dopamine and the like. With the continuous and intensive research, antitumor small molecules with pyrrolopyridine as a skeleton have been approved by the FDA for marketing. Vemurafenib (Vemurafenib), a BRAF selective inhibitor with half-inhibitory concentration of 31nM, was FDA approved in 2011, month 8 for the treatment of advanced metastatic or surgically unresectable melanoma. GSK1070916 is an ATP competitive aurora kinase inhibitor taking pyrrolopyridine as a framework, ki of aurora kinase B and aurora kinase C are respectively 0.38+/-0.29 nM and 1.5+/-0.4 nM, and a tumor proliferation inhibition activity evaluation experiment shows that the half inhibition concentration of GSK1070916 on lung cancer cells is 7nM, and proliferation of tumor cells is effectively inhibited. Xenograft tumor experiments show that GSK1070916 can inhibit phosphorylation of histone H3 of human colon cancer Colo205 and HL-60, and the small molecule is in clinical phase I experiments at present.
The anti-tumor target PAK5 is proved to play an important role in the aspects of invasion and metastasis, proliferation, survival, apoptosis and the like of tumor cells, so that the research and development of inhibitors thereof are considered as effective strategies for tumor treatment, while class II PAKs inhibitors are mostly in the biological test research stage. However, based on a great deal of research results of molecular biology and structural biology, the discovery and development of class II PAKs inhibitors, especially selective class II PAKs inhibitors, are in a rapid development stage at present, and a new generation of antitumor drugs, especially protein crystal structures of PAK5, are expected to be analyzed, and it is believed that the development of PAK5 inhibitors will be imperative.
Therefore, how to provide a protein kinase inhibitor capable of effectively inhibiting PAK5 activity is a problem to be solved by those skilled in the art.
Disclosure of Invention
The application aims to obtain a class of pyrrolopyridine compounds based on serine/threonine kinase PAK5 and by utilizing a fragment-based drug design principle on the basis of the prior art, and pharmacological experiments prove that the compounds have good PAK5 inhibition activity and particularly have stronger anti-tumor activity on tumor cells such as kidney cancer cells, liver cancer cells, colorectal cancer cells, breast cancer cells and the like.
The application also aims to provide a preparation method of the pyrrolopyridine compound.
Another object of the present application is to provide a pharmaceutical use of the pyrrolopyridines described above, in particular to a pharmaceutical use for preparing a medicament for diseases related to serine/threonine kinase PAK 5; wherein the serine/threonine kinase PAK5 related diseases are colorectal cancer, liver cancer, gastric cancer, breast cancer, renal cancer or cervical cancer.
The technical scheme of the application is as follows:
a compound, isomer, hydrate, solvate or pharmaceutically acceptable salt thereof, of formula I:
wherein,,
R 1 represents hydrogen, C 1 -C 4 Alkyl, C 1 -C 4 Alkoxy, acryl or t-butoxycarbonyl;
R 2 represents hydrogen, hydroxy, C 1 -C 4 Alkyl, C 1 -C 4 Alkoxy, halogen or trifluoromethyl;
L 1 represents-CH 2 -、-CO-NH-、-CO-NH-CH 2 -C (OH) -, vinyl, ethynyl, nitrogen atom, hydrazide or sulfonyl hydrazide;
n represents 0, 1 or 2;
ring a represents piperidine, piperazine, morpholine, pyrrole or furan;
ring B represents phenyl or a nitrogen-containing five-membered heterocycle;
ring C represents phenyl, naphthyl or biphenyl;
ring D represents an oxygen atom, phenyl, cyclopentane, cyclohexane, pyridine or pyrimidine.
In one placeIn a preferred embodiment, R 1 Represents hydrogen, methyl, ethyl, methoxy or ethoxy.
In a more preferred embodiment, R 1 Represents methyl.
In a preferred embodiment, R 2 Represents hydrogen, hydroxy, methyl, ethyl, methoxy or ethoxy.
In a more preferred embodiment, R 2 Represents hydrogen, hydroxy or methoxy.
In a preferred embodiment, L 1 represents-CH 2 -、-CO-NH-、-CO-NH-CH 2 -C (OH) -, vinyl, ethynyl or hydrazide groups.
In a more preferred embodiment, L 1 represents-CH 2 -、-CO-NH-、-CO-NH-CH 2 -C (OH) -or ethynyl.
In a preferred embodiment, n represents 0 or 1.
In a more preferred embodiment, n represents 0.
In a preferred embodiment, ring a represents piperidine, piperazine or morpholine.
In a more preferred embodiment, ring a represents piperidine.
In a preferred embodiment, ring B represents phenyl, pyrazole, oxazole, thiazole, imidazole or pyrrole.
In a more preferred embodiment, ring B represents pyrazole.
In a preferred embodiment, ring C represents phenyl.
In a preferred embodiment, ring D represents an oxygen atom, phenyl, cyclopentane, cyclohexane, pyridine or pyrimidine.
In a more preferred embodiment, ring D represents an oxygen atom, phenyl, cyclopentane, cyclohexane or pyridine.
Further, R 1 Represents methyl; r is R 2 Represents hydrogen, hydroxy or methoxy; l (L) 1 represents-CH 2 -、-CO-NH-、-CO-NH-CH 2 -C (OH) -or ethynyl; n represents 0; ring a represents piperidine; ring B represents pyrazole; ring C represents phenyl; ring D represents an oxygen atom, phenyl, cyclopentane, cyclohexane or pyridine.
Still further, a compound, isomer, hydrate, solvate or pharmaceutically acceptable salt thereof, as described in formula I, wherein said compound is selected from the group consisting of:
in a preferred embodiment, when ring A represents piperidine, ring B represents pyrazole, ring C represents phenyl, L 1 Represents ethynyl and R 1 When representing methyl, the preparation method of the compound shown in the general formula I comprises the following steps:
the intermediates or target compounds mentioned in the present application can be purified by conventional isolation techniques and, if desired, converted into addition salts with pharmaceutically acceptable acids.
Unless otherwise indicated, the following terms used in the specification and claims have the meanings discussed below:
"pharmaceutically acceptable salts" means those salts which retain the biological effectiveness and properties of the parent compound. Such salts include:
(1) Salified with acids obtained by reaction of the free base of the parent compound with inorganic acids including hydrochloric acid, hydrobromic acid, nitric acid, phosphoric acid, metaphosphoric acid, sulfuric acid, sulfurous acid, perchloric acid, and the like, or with organic acids including acetic acid, trifluoroacetic acid, propionic acid, acrylic acid, caproic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, oxalic acid, (D) or (L) malic acid, fumaric acid, maleic acid, ascorbic acid, camphoric acid, benzoic acid, hydroxybenzoic acid, gamma-hydroxybutyric acid, methoxybenzoic acid, phthalic acid, methanesulfonic acid, ethanesulfonic acid, naphthalene-1-sulfonic acid, naphthalene-2-sulfonic acid, p-toluenesulfonic acid, salicylic acid, tartaric acid, citric acid, lactic acid, cinnamic acid, dodecylsulfuric acid, gluconic acid, glutamic acid, aspartic acid, stearic acid, mandelic acid, succinic acid, glutaric acid, malonic acid, and the like.
(2) The acidic proton present in the parent compound is replaced by a metal ion or a salt formed by complexation with an organic base such as alkali metal ion, alkaline earth metal ion or aluminum ion, and an organic base such as ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine, quinine, etc.
"pharmaceutical composition" means that one or more of the compounds of the present application, or a pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof, is admixed with another chemical ingredient, such as a pharmaceutically acceptable carrier. The purpose of the pharmaceutical composition is to facilitate the process of administration to animals.
"pharmaceutically acceptable carrier" or "pharmaceutically acceptable carrier" refers to an inactive ingredient in a pharmaceutical composition that does not cause significant irritation to the organism and does not interfere with the biological activity and properties of the compound being administered, such as, but not limited to: calcium carbonate, calcium phosphate, various sugars (e.g., lactose, mannitol, etc.), starch, cyclodextrin, magnesium stearate, cellulose, magnesium carbonate, acrylic or methacrylic polymers, gelatin, water, polyethylene glycol, propylene glycol, ethylene glycol, castor oil or hydrogenated castor oil or polyethoxylated hydrogenated castor oil, sesame oil, corn oil, peanut oil, and the like.
"alkyl" means a saturated aliphatic radical of 1 to 20 carbon atoms and includes straight and branched chain groups (the numerical ranges mentioned herein, e.g., "1 to 20", refer to such groups, which in this case are alkyl groups, which may contain 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., up to and including 20 carbon atoms). More preferably, the alkyl group is a medium size alkyl group having 1 to 10 carbon atoms, such as methyl, ethyl, propyl, 2-propyl, n-butyl, isobutyl, t-butyl, pentyl, and the like. Preferably, the alkyl group is a lower alkyl group having 1 to 8 or 1 to 6 carbon atoms, such as methyl, ethyl, propyl, 2-propyl, n-butyl, isobutyl, tert-butyl, etc. Alkyl groups may be substituted or unsubstituted. When substituted alkyl, the substituent is preferably one or more, more preferably 1 to 3, most preferably 1 or 2 substituents.
The nitrogen-containing five-membered heterocycle represents a single ring of 5 ring atoms containing one, two, three or four N heteroatoms, the remaining ring atoms being C, for example pyrazole, oxazole, thiazole, imidazole or pyrrole.
"hydroxy" means an-OH group.
"trifluoromethyl" means-CF 3 A group.
"alkoxy" means-O- (unsubstituted alkyl) and-O- (unsubstituted cycloalkyl). Representative examples include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, cyclopropoxy, cyclobutoxy, cyclopentoxy, cyclohexyloxy, and the like.
"halogen" means fluorine, chlorine, bromine or iodine, preferably fluorine or chlorine.
The application provides a pharmaceutical composition, which takes the compound, isomer, hydrate, solvate or pharmaceutically acceptable salt thereof as an active ingredient or a main active ingredient, and is assisted with a pharmaceutically acceptable carrier.
The compound, isomer, hydrate, solvate or pharmaceutically acceptable salt thereof can be applied to the preparation of medicines related to serine/threonine kinase PAK5, wherein the diseases related to the serine/threonine kinase PAK5 are colorectal cancer, liver cancer, gastric cancer, breast cancer, kidney cancer or cervical cancer.
By adopting the technical scheme of the application, the advantages are as follows:
1. the compound has better inhibition effect on human colon cancer HCT-116 cells, human kidney clear cell carcinoma 786O cells, human breast cancer MCF-7 cells and human liver cancer HepG2 cells. Wherein, compound III 5 Half of the inhibition concentration of the four tumor cells is at nanomole level, and the inhibition effect is superior to that of Sunitinib (Sunitinib); normal cell human renal cortex proximal tubular epithelium at 1 μm concentrationThe survival rate of the cell HK-2 is close to 80%, which shows that the derivative taking the pyrrolopyridine as the skeleton has a strong inhibition effect on tumor cells and good biological safety.
2. The inhibition of the PAK5 protein by the target compound was evaluated using a Homogeneous Time Resolved Fluorescence (HTRF) STK-S2 kit (62 ST2PEB, cisbio), overall at nanomolar level. Wherein Compound III 5 IC for PAK5 50 8.+ -. 0.40nM.
3. HepG2 xenograft tumor nude mice experiments prove that the target compound III 5 The tumor volume and tumor weight of the group are obviously smaller than those of the positive control medicine group, and the tumor inhibition rate reaches 91.21 percent
4. After the end of the administration, HE-stained sections of heart, liver, spleen, lung and kidney tissues of nude mice showed no pathological structural changes, indicating that Compound III 5 Has better biological safety.
Drawings
FIG. 1 is the effect of compound III5 on the survival of normal cells HK-2; wherein, compound III 5 At a concentration of 1. Mu.M, the survival rate of normal cells human kidney cortex proximal tubular epithelial cells HK-2 cells is approximately 80%;
FIG. 2 is the inhibition of HepG2 nude mice engrafting tumor model by Compound III 5; fig. 2a is a photograph of tumor of nude mice obtained by peeling each treatment group after 16 days of administration; fig. 2B is a graph showing the change in tumor volume after administration; FIG. 2C shows the change in tumor weight after administration; target compound III 5 The tumor volume and tumor weight of the group are obviously smaller than those of the positive control medicine group, and the tumor inhibition rate reaches 91.21%;
FIG. 3 is a graph showing the change in body weight of nude mice during administration; the high, medium and low dose groups of the compound III5 have smaller influence on the weight of the nude mice, which indicates that the compound has smaller toxicity;
FIG. 4 is the effect of compound III5 on normal tissues of nude mice; wherein, after the end of the administration, HE staining sections of heart, liver, spleen, lung and kidney tissues of nude mice have no pathological structural changes, which indicates that the toxicity of the compound III5 is smaller
Detailed Description
The following description of the technical solutions in the embodiments of the present application will be clear and complete, and it is obvious that the described embodiments are only some embodiments of the present application, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the application without making any inventive effort, are intended to be within the scope of the application.
In view of the fact that the pyrrolopyridines protected by the application are more classified, the applicant only enumerates some compounds in the pyrrolopyridines, and other compounds can achieve the following technical effects.
EXAMPLE 1 Compound III 5 The structural formula and the detailed preparation method are as follows:
the preparation method comprises the following steps:
synthesis of tert-butyl 4- (4- (1-hydro-pyrrolo [2,3-b ] pyridin-5-yl) -1H-pyrazol-1-yl) piperidine-1-carboxylate (10)
The raw material 5-bromo-7-azaindole (3.94 g,20 mmol), 1- (1-t-butoxycarbonyl) -piperidinyl-4-boronate-pyrazole (8.67 g,24 mmol) was weighed into a eggplant-shaped bottle, and Pd (PPh) 3 ) 4 (1.2g,1mmol)，Na 2 CO 3 (4.24 g,40 mmol) was added to 10mL of the mixed solvent (dioxane: water=4:1), dissolved with stirring, nitrogen-protected, reacted at 80℃and monitored by TLC (DCM: meOH=30:1). After the reaction was completed, the reaction solution was extracted, the organic phase was dried, concentrated, sand-packed on silica gel, and separated by column chromatography (DCM: meoh=50:1) to give a yellow solid. 1 H NMR(400MHz,CDCl 3 )δ9.39(s,1H),8.43(s,1H),8.00(d,J＝2.0Hz,1H),7.80(d,J＝2.4Hz,1H),7.68(d,J＝1.2Hz,1H),7.33–7.30(m,1H),4.34–4.31(m,1H),2.97–2.87(m,2H),2.22–2.16(m,2H),2.01–1.96(m,2H),1.86–1.80(m,2H),1.48(s,9H).
Synthesis of tert-butyl 4- (4- (3-iodo-1-hydro-pyrrolo [2,3-b ] pyridin-5-yl) -1H-pyrazol-1-yl) piperidine-1-carboxylate (11)
Raw material 10 (3.67 g,10 mmol) is weighed into a eggplant-shaped bottle, dichloromethane is dissolved and placed at room temperature, stirring is started, after 5min, N-iodosuccinimide (2.7 g,12 mmol) is weighed in batches, the batch is put into the bottle for 30min, stirring is continued, TLC monitoring (DCM: meOH=30:1) is carried out, after the reaction is finished, the reaction is quenched, ethyl acetate is extracted, an organic phase is dried and concentrated, silica gel sand is made, column chromatography is carried out, and yellow solid is separated. 1 H NMR(400MHz,DMSO-d 6 )δ12.07(s,1H),8.55(d,J＝2.0Hz,1H),8.43(s,1H),8.00(s,1H),7.82(d,J＝2.0Hz,1H),7.70(d,J＝2.5Hz,1H),4.42–4.33(m,1H),4.09–4.02(m,2H),2.99–2.89(m,2H),2.11–2.05(m,2H),1.86–1.78(m,2H),1.43(s,9H).
Synthesis of tert-butyl 4- (4- (3-iodo-1-methyl-benzenesulfonyl-1-hydro-pyrrolo [2,3-b ] pyridin-5-yl) -1H-pyrazol-1-yl) piperidine-1-carboxylate (12)
Raw material 11 (4.93 g,10 mmol) is weighed and put into a eggplant-shaped bottle, 10mL of DMF is dissolved, the mixture is placed in an ice bath, stirring is started, sodium hydride (287.97 mg,12 mmol) is weighed and put into the bottle in batches for 30 minutes, stirring is continued for 1 hour, p-toluenesulfonyl chloride (2.29 g,12 mmol) is weighed and put into the bottle, stirring is continued, TLC monitoring (DCM: meOH=50:1) is continued, after the reaction is finished, the reaction solution is transferred into ice water, stirring is carried out for 30 minutes, suction filtration is carried out, and filter residues are washed with water, so that off-white solid is obtained. 1 H NMR(400MHz,DMSO-d 6 )δ8.65(d,J＝2.0Hz,1H),8.45(s,1H),8.07(s,1H),8.02(d,J＝1.4Hz,1H),7.98(s,1H),7.96(s,1H),7.86(d,J＝2.1Hz,1H),7.58(d,J＝8.3Hz,1H),7.42–7.35(m,1H),4.37–4.26(m,1H),2.48–2.44(m,2H),2.37–2.34(m,2H),2.28(s,3H),2.04–1.97(m,2H),1.79–1.73(m,2H),1.37(s,9H).
Synthesis of 3-iodo-5- (1- (piperidin-4-yl) -1H-pyrazol-4-yl) -1-methyl-benzenesulfonyl-1H-pyrrolo [2,3-b ] pyridine (13)
Raw material 12 (3.24 g,5 mmol) was weighed into a eggplant-shaped bottle, dissolved in methanol, stirred at room temperature, hydrochloric acid was taken and slowly added dropwise to excess, TLC monitoring (DCM: meOH=20:1) was carried out, and after the reaction was completed, the reaction solution was concentrated to prepare the next reaction.
Synthesis of 3-iodo-5- (1- (1-methyl-piperidin-4-yl) -1H-pyrazol-4-yl) -1-methyl-benzenesulfonyl-1H-pyrrolo [2,3-b ] pyridine (14)
Dissolving the raw material 13 with methanol, adding a stirrer, stirring at room temperature, adding catalytic amount of acetic acid, taking 7mL of formaldehyde solution, slowly adding dropwise, stirring at room temperature for 2 hours, directly extracting the reaction liquid, drying the organic phase, and concentrating to obtain white solid.
The white solid was taken and put into a 25mL eggplant-shaped bottle, dissolved in methanol, and sodium cyanoborohydride (378 mg,1.2 mmol) was weighed in portions, added in portions, stirred at room temperature, monitored by TLC (DCM: meoh=20:1), quenched by reaction solution after the reaction was completed, extracted, dried and concentrated the organic phase, sand-made on silica gel, and separated by column chromatography (MeOH: dcm=50:1) to give a white solid. 1 H NMR(400MHz,CDCl 3 )δ8.57(d,J＝2.1Hz,1H),8.10(d,J＝8.4Hz,2H),7.87(s,1H),7.81(s,1H),7.75(s,1H),7.67(d,J＝2.1Hz,1H),7.31(d,J＝7.8Hz,2H),4.23–4.17(m,1H),3.06–3.00(m,2H),2.39(s,3H),2.38(s,3H),2.23–2.20(m,2H),2.14–2.09(m,2H),1.30–1.24(m,2H).
Synthesis of 1- ((3-bromo-phenyl) ethynyl) cyclohexyl-1-ol (III) 5a )
Meta-bromoiodobenzene (1.41 g,5 mmol), 1-ethynyl cyclohexanol (745.1 mg,6 mmol), cuprous iodide (95.23 mg,0.5 mmol), triethylamine (1.01 g,10 mmol) and [1,1' -bis (diphenylphosphino) ferrocene were weighed out]Palladium dichloride (183mg, 0.25 mmol), 5mL tetrahydrofuran was dissolved, nitrogen protected, and reacted at 70 ℃. TLC monitoring (PE: EA=5:1), after the reaction is finished, extracting the reaction liquid, drying and concentrating the organic phase, preparing sand by silica gel, and separating by column chromatography (PE: EA=5:1) to obtain yellow solid. 1 H NMR(400MHz,CDCl 3 )δ7.58(t,J＝1.8Hz,1H),7.44(m,1H),7.35(m,1H),7.17(t,J＝8.0Hz,1H),2.08–1.95(m,4H),1.64–1.56(m,6H).
Synthesis of 1- (3-boronate-phenyl) -ethynyl-cyclohexan-1-ol (III) 5b )
Weighing III 5a (1.4 g,5 mmol), pinacol biborate (1.52 g,6 mmol), potassium acetate (981.42 mg,10 mmol), [1,1' -bis (diphenylphosphino) ferrocene]Palladium dichloride (183mg, 0.25 mmol), 5mL DMSO was dissolved, nitrogen protected, and reacted at 90 ℃. TLC monitoring (PE: EA=5:1), after the reaction is finished, extracting the reaction liquid, drying and concentrating the organic phase, preparing sand by silica gel, and separating by column chromatography (PE: EA=5:1) to obtain a white solid. 1 H NMR(400MHz,CDCl 3 )δ7.91(s,1H),7.76(m,1H),7.54(m,1H),7.34(t,J＝7.5Hz,1H),1.78–1.72(m,4H),1.63–1.54(m,6H),1.37(s,12H).
1- ((3- (5- (1- (1-methyl-piperidin-4-yl) -1H-pyrazol-4-yl) -1H-pyrrole [2, 3-b)]Synthesis of pyridin-3-yl) phenyl-ethynyl) cyclohexyl-1-ol (III) 5 )
Weighing raw material III 5b (200 g,0.35 mmol), 14 (140 mg,0.14 mmol), sodium carbonate (75 mg,0.70 mmol) was charged into a 100ml round bottom flask, and tetrakis (triphenylphosphine) palladium (40 mg,0.035 mmol) was weighed into the reaction flaskDissolving with a mixed solvent (dioxane: water=4:1), protecting with nitrogen, reacting at 80 ℃, monitoring by TLC (DCM: meOH=20:1), extracting the reaction liquid by ethyl acetate after the reaction is finished, drying and concentrating an organic phase, preparing sand by silica gel, and separating by column chromatography (DCM: meOH=40:1) to obtain pale white solid.
The product from the previous step (100 mg,0.15 mmol), potassium carbonate (23 mg,0.15 mmol) was taken into a 50ml round bottom flask, dissolved in a mixed solvent (methanol: water=4:1), refluxed for 5h at 60 ℃, monitored by tlc (DCM: meoh=15:1), after the reaction was completed, the reaction solution was extracted with ethyl acetate, the organic phase was dried, concentrated, silica gel was made up, and separated by column chromatography (DCM: meoh=40:1) to give a white solid.
Compound III 5 The product was a white solid, 0.512g, 65.07% yield. 1 H NMR(400MHz,MeOD)δ8.50(d,J＝2.0Hz,1H),8.38(d,J＝2.0Hz,1H),8.20(s,1H),7.92(s,1H),7.77–7.68(m,3H),7.46(t,J＝7.7Hz,1H),7.35(m,J＝7.7Hz,1H),4.25(m,J＝5.1Hz,1H),3.23(s,1H),3.05(d,2H),2.37(s,3H),2.28(t,J＝3.3Hz,2H),2.23–2.11(m,4H),2.02(d,2H),1.78(d,J＝4.3Hz,1H),1.69(t,3H),1.63(d,J＝7.2Hz,1H),1.35(s,1H),1.30(s,1H).HRMS(ESI)m/z[M-H] + :478.2607,found:478.2593.Retention time 2.612min,HPLC purity＝98.255％.
EXAMPLE 2 Compound III 4 The structural formula and the detailed preparation method are as follows:
the preparation method comprises the following steps:
wherein III 4a Referring to example 1, the 1-ethynyl cyclohexanol was replaced with 1-ethynyl cyclopentanol, and the synthetic route was as follows:
m-bromoiodobenzene (1.41 g,5 mmol), 1-ethynyl cyclopentanol (660.94 mg,6 mmol), cuprous iodide (95.23 mg,0.5 mmol), triethylamine (1.01 g,10 mmol), [1,1' -bis (diphenylphosphino) ferrocene ], palladium dichloride (183mg, 0.25 mmol), tetrahydrofuran were weighed out, nitrogen substitution for 5min, and reacted overnight at 70 ℃. TLC detection (PE: EA=5:1) until the reaction of the starting materials is complete, extraction of the reaction solution, drying and concentration of the organic phase, sand making with silica gel, column chromatography separation (PE: EA=5:1) to obtain a yellow solid.
Wherein III 4b With reference to example 1, the synthetic route is as follows:
weighing III 4a (1.2 g,5 mmol), pinacol biborate (1.52 g,6 mmol), potassium acetate (981.42 mg,10 mmol), [1,1' -bis (diphenylphosphino) ferrocene]Palladium dichloride (183mg, 0.25 mmol), 5mL DMSO was dissolved, nitrogen protected, and reacted at 90 ℃. TLC monitoring (PE: EA=5:1), after the reaction is finished, extracting the reaction liquid, drying and concentrating the organic phase, preparing sand by silica gel, and separating by column chromatography (PE: EA=5:1) to obtain a white solid. 1 H NMR(400MHz,CDCl 3 )δ7.89(d,J＝1.6Hz,1H),7.72(m,1H),7.50(m,1H),7.30(t,J＝7.6Hz,1H),2.03(m,4H),1.86–1.84(m,2H),1.81–1.74(m,2H),1.34(s,12H).
Wherein III 4 With reference to example 1, the synthetic route is as follows:
weighing raw material III 4b (198mg, 0.63 mmol), 14 (300 mg,0.53 mmol), sodium carbonate (112 mg,1.06 mmol) were put into a 100ml eggplant-shaped bottle, tetrakis (triphenylphosphine) palladium (31 mg,0.026 mmol) was weighed into a reaction bottle, dissolved in a mixed solvent (dioxane: water=4:1), reacted at 80℃under nitrogen, monitored by TLC (DCM: meOH=20:1), and after the reaction was completed, the reaction solution was extracted with ethyl acetate, the organic phase was dried, concentrated, sand-made with silica gel, and separated by column chromatography (DCM: meOH=40:1) to give a white solid.
The product from the previous step (100 mg,0.16 mmol), potassium carbonate (72 mg,0.48 mmol) was put into a 50ml round bottom flask, dissolved in a mixed solvent (methanol: water=4:1), refluxed for 5h at 60 ℃, monitored by tlc (DCM: meoh=15:1), and after the reaction was completed, the reaction solution was extracted with ethyl acetate, the organic phase was dried, concentrated, silica gel was made into sand, and column chromatography was separated (DCM: meoh=40:1) to give white solid III 4 。
1- ((3- (5- (1- (1-methylpiperidin-4-yl) -1H-pyrazol-4-yl) -1H-pyrrole [2, 3-b)]Pyridin-3-yl) phenyl ethynyl) cyclopentan-1-ol (III) 4 ) White solid, 0.37g, yield 50.07%, 1 H NMR(400MHz,MeOD)δ8.48(d,J＝2.0Hz,1H),8.36(d,J＝2.0Hz,1H),8.18(s,1H),7.91(s,1H),7.72-7.65(m,3H),7.43(t,J＝7.7Hz,1H),7.32(d,J＝7.7Hz,1H),3.21(s,1H),3.07-2.99(m,2H),2.35(s,3H),2.31-2.22(m,2H),2.19-2.12(m,4H),2.03(d,J＝6.5Hz,4H),1.90-1.76(m,4H).HRMS(ESI)m/z[M-H] + :464.2456,found:464.2430.Retention time 2.55min,HPLC purity＝97.681％.
EXAMPLE 3 Compound III 1 The structural formula and the detailed preparation method are as follows:
compound 14 is prepared in the same manner as in example 1, wherein III 1b With reference to example 1, the synthetic route is as follows:
(3-bromophenyl) methanol (935.18 mg,5 mmol), pinacol biborate (1.52 g,6 mmol), potassium acetate (981.42 mg,10 mmol), [1,1' -bis (diphenylphosphino) ferrocene ] palladium dichloride (183mg, 0.25 mmol), 5mL DMSO dissolved, nitrogen blanket, and reacted at 90℃were weighed out. TLC monitoring (PE: EA=5:1), after the reaction is finished, extracting the reaction liquid, drying and concentrating the organic phase, preparing sand by silica gel, and separating by column chromatography (PE: EA=5:1) to obtain a white solid.
Wherein III 1 With reference to example 1, the synthetic route is as follows:
weighing raw material III 1b (150 mg,0.64 mmol), 14 (300 mg,0.53 mmol), sodium carbonate (112 mg,1.06 mmol) were charged into a 100ml round bottom flask, tetrakis (triphenylphosphine) palladium (31 mg,0.026 mmol) was weighed into a reaction flask, dissolved in a mixed solvent (dioxane: water=4:1), reacted at 80 ℃ under nitrogen, monitored by TLC (DCM: meoh=20:1), and after the reaction was completed, the reaction solution was extracted with ethyl acetate, the organic phase was dried, concentrated, silica gel was made into sand, and column chromatography was separated (DCM: meoh=40:1) to give a pale yellow solid.
The product from the previous step (150 mg,0.28 mmol), potassium carbonate (129 mg,0.84 mmol) was taken into a 50ml round bottom flask and dissolved in a mixed solvent (methanol: water=4:1), reacted at 60℃under reflux for 5h, monitored by TLC (DCM: meOH=15:1). After the reaction was completed, ethyl acetate was extracted, the organic phase was dried, concentrated, silica gel was made up, and column chromatographed (DCM: meoh=40:1) to give a white solid powder.
(3- (5- (1- (1-methylpiperidin-4-yl) -1H-pyrazol-4-yl) -1H-pyrrolo [2, 3-b)]Pyridin-3-yl) phenyl methanol (III) 1 ) White solid powder, 0.131g, yield 68.11%, 1 H NMR(400MHz,CDCl 3 )δ10.74(s,1H),8.54(s,1H),8.24(d,J＝1.8Hz,1H),7.97(s,1H),7.78(d,J＝0.7Hz,1H),7.66(d,J＝2.0Hz,1H),7.61–7.57(m,1H),7.50–7.45(m,2H),7.33(d,J＝7.7Hz,1H),4.80(s,2H),4.32–4.26(m,1H),2.25–2.17(m,4H),1.87(s,4H).HRMS(ESI)m/z[M-H] + :386.1986,found:386.2004.Retention time 2.091min,HPLC purity＝98.713％.
EXAMPLE 4 Compound III 2 The structural formula and the detailed preparation method are as follows:
the preparation method comprises the following steps:
the procedure for the preparation of compound 14 was as in example 1, and the synthetic route was as follows:
(3-bromophenyl) methanol (935.18 mg,5 mmol), pinacol biborate (1.52 g,6 mmol), potassium acetate (981.42 mg,10 mmol), [1,1' -bis (diphenylphosphino) ferrocene ] palladium dichloride (183mg, 0.25 mmol), 5mL DMSO dissolved, nitrogen blanket, and reacted at 90℃were weighed out. TLC monitoring (PE: EA=5:1), after the reaction is finished, extracting the reaction liquid, drying and concentrating the organic phase, preparing sand by silica gel, and separating by column chromatography (PE: EA=5:1) to obtain a white solid.
Wherein III 2b With reference to example 1, the synthetic route is as follows:
3-boronic acid ester-benzoic acid (496.17 mg,2 mmol), 3-amino-2-methoxy-pyridine (297.94 mg,2.4 mmol), HATU (564.65 mg,2,4 mmol), DIPEA (516.97 mg,4 mmol) were weighed out, 3mL DMF was dissolved, stirred at room temperature, monitored by TLC (PE: ea=1:1), after the reaction was completed, the reaction solution was extracted, the organic phase was dried, concentrated, sand was made on silica gel, and column chromatography was carried out (PE: ea=1:1) to obtain a white solid. 1 H NMR(400MHz,CDCl 3 ):δ8.72(d,J＝9.6Hz,1H),8.42(s,1H),8.28(s,1H),7.99(d,J＝9.1Hz,2H),7.90(d,J＝1.9Hz,1H),7.51(t,J＝7.6Hz,1H),6.95(dd,J＝7.8,5.0Hz,1H),4.05(s,3H),3.97(s,1H),1.36(s,12H).
Wherein III 2 With reference to example 1, the synthetic route is as follows:
weighing raw material III 2b (227 mg,0.64 mmol), 14 (300 mg,0.53 mmol), sodium carbonate (112 mg,1.06 mmol) was charged into a 100ml round bottom flask, tetrakis (triphenylphosphine) palladium (31 mg,0.026 mmol) was weighed into a reaction flask, dissolved in a mixed solvent (dioxane: water=4:1), and nitrogen gasProtection, reaction at 80 ℃, TLC monitoring (DCM: meoh=20:1), extraction of the reaction with ethyl acetate, drying of the organic phase, concentration, silica gel trituration, column chromatography separation (DCM: meoh=40:1) to give a pale yellow solid.
The product from the previous step (150 mg,0.23 mmol), potassium carbonate (106 mg,0.69 mmol) was taken into a 50ml round bottom flask, dissolved in a mixed solvent (methanol: water=4:1), refluxed for 5h at 60 ℃, monitored by TLC (DCM: meOH=15:1), after the reaction was completed, the reaction solution was extracted with ethyl acetate, the organic phase was dried, concentrated, silica gel was made up, and separated by column chromatography (DCM: meOH=40:1) to give a white solid powder III 2 。
N- (2-methoxypyridin-3-yl) -3- (5- (1- (1-methylpiperidin-4-yl) -1H-pyrazol-4-yl) -1H-pyrrolo [2,3-b]Pyridin-3-yl) benzamide (III) 2 ) White solid, 0.62g, yield 61.37%, 1 H NMR(400MHz,CDCl 3 )δ10.75(s,1H),8.78(dd,J＝7.8,1.7Hz,1H),8.63(d,J＝2.0Hz,1H),8.49(s,1H),8.30(d,J＝2.0Hz,1H),8.20(t,J＝1.8Hz,1H),8.06(s,1H),7.92–7.76(m,4H),7.65–7.60(m,2H),6.98(dd,J＝7.8,5.0Hz,1H),4.04(s,3H),2.51(s,3H),2.36–2.14(m,4H).HRMS(ESI)m/z[M-H] + :506.2304,found:506.2322.Retention time 2.439min,HPLC purity＝98.867％.
EXAMPLE 5 Compound III 3 The structural formula and the detailed preparation method are as follows:
the preparation method comprises the following steps:
compound 14 is prepared in the same manner as in example 1, wherein III 3a With reference to example 1, the synthetic route is as follows:
(3-bromophenyl) formic acid (1.05 g,5 mmol), pinacol biborate (1.52 g,6 mmol), potassium acetate (981.42 mg,10 mmol) and [1,1' -bis (diphenylphosphino) ferrocene ] palladium dichloride (183mg, 0.25 mmol) were weighed out, 5mL DMSO was dissolved, nitrogen protected, and reacted at 90 ℃. TLC monitoring (PE: EA=5:1), after the reaction is finished, extracting the reaction liquid, drying and concentrating the organic phase, preparing sand by silica gel, and separating by column chromatography (PE: EA=5:1) to obtain yellow solid.
Wherein III 3b With reference to example 1, the synthetic route is as follows:
3-boronic acid ester-benzoic acid (496.17 mg,2 mmol), 2-amino-1- (329.24 mg,2.4 mmol), HATU (564.65 mg,2,4 mmol), DIPEA (516.97 mg,4 mmol), 3mL DMF were weighed out, stirred at room temperature, monitored by TLC (PE: ea=1:1), after the reaction was completed, the reaction solution was extracted, the organic phase was dried, concentrated, silica gel was made up, and column chromatography was separated (PE: ea=1:1) to obtain a white solid. 1 H NMR(400MHz,CDCl 3 ):δ8.13(t,J＝1.4Hz,1H),7.98–7.91(m,2H),7.47–7.34(m,6H),6.91(s,1H),3.94–3.86(m,1H),3.56–3.48(m,1H),1.35(s,12H).
Wherein III 2 With reference to example 1, the synthetic route is as follows:
weighing raw material III 3b (235 mg,0.64 mmol), 14 (300 mg,0.53 mmol), sodium carbonate (112 mg,1.06 mmol) were charged into a 100ml round bottom flask, tetrakis (triphenylphosphine) palladium (31 mg,0.026 mmol) was weighed into a reaction flask, dissolved in a mixed solvent (dioxane: water=4:1), reacted at 80 ℃ under nitrogen, monitored by TLC (DCM: meoh=20:1), and after the reaction was completed, the reaction solution was extracted with ethyl acetate, the organic phase was dried, concentrated, silica gel was made into sand, and column chromatography was separated (DCM: meoh=40:1) to give a pale yellow solid.
The product from the previous step (150 mg,0.22 mmol), potassium carbonate (101 mg,0.66 mmol) was taken into a 50ml round bottom flask, dissolved in a mixed solvent (methanol: water=4:1), refluxed for 5h at 60℃and monitored by TLC(DCM: meoh=15:1), after the reaction was completed, the reaction was extracted with ethyl acetate, the organic phase was dried, concentrated, silica gel was prepared as a sand, and column chromatographed (DCM: meoh=40:1) to give white solid powder III 3 。
(S) -N- (2-hydroxy-2-phenethyl) -3- (5- (1- (1-methylpiperidin-4-yl) -1H-pyrazol-4-yl) -1H-pyrrolo [2,3-b]Pyridin-3-yl) benzamide (III) 3 ) White solid, 0.33g, yield 51.98%, 1 H NMR(400MHz,CDCl 3 )δ8.48(d,J＝2.0Hz,1H),8.36(d,J＝2.0Hz,1H),8.18(s,1H),7.92(s,1H),7.71–7.65(m,3H),7.44(t,J＝7.7Hz,1H),7.32(d,J＝7.7Hz,1H),7.17–7.11(m,5H),5.36(s,1H),5.34(s,1H),4.30(t,J＝6.7Hz,1H),3.49(s,2H),2.54(d,J＝16.0Hz,4H),2.00(d,J＝6.6Hz,4H).HRMS(ESI)m/z[M-H] + :519.2508,found:519.2521.Retention time 2.105min,HPLC purity＝98.397％.
the following are some of the pharmacological tests and results of representative compounds of the present application:
1. CCK-8 method for detecting proliferation inhibition effect of target compound on tumor cells
Tumor cells were digested with pancreatin, counted by cell counting plate, and 36 wells around 96 well plate were filled with 100 μl PBS to prevent evaporation of water, and 5000-10000 cells were plated per well. After cell attachment, complete medium containing specific concentration of compound is added, and the mixture is placed at 37 ℃ and 5% CO 2 The cells were cultured in a cell incubator for 48 hours. After 48h, the medium in the 96-well plate was discarded, and 90. Mu.L of complete medium and 10. Mu.L of CCK-8 solution were added per well in the dark and mixed well. The 96-well plates were placed in a cell incubator for incubation for 0.5,1,2,4h. The wavelength of the enzyme-labeled instrument is set to 450nm to measure OD value, and the experiment is repeated three times. Relative inhibition = (OD control-OD experimental)/(OD control-OD blank) ×100%. Calculating IC according to the koidz algorithm 50 The value, i.e. the drug concentration with a cell growth inhibition of 50%. Specific proliferation inhibition effects are shown in table 1.
Effect of the compounds of Table 1 on the antiproliferative capacity of different tumor cell lines
The experimental results of CCK-8 in Table 1 show that the compound provided by the application has obvious proliferation inhibition effect on human colorectal cancer cells HCT-116, human breast cancer cells MCF-7, human kidney cancer cells 786O and human liver cancer cells HepG 2.
Wherein Compound III 2 Half-maximal inhibitory concentrations for MCF-7 cells and 786O cells were 0.44 and 0.83 μm, respectively; compound III 4 Half inhibition concentration for 786O cells was 0.95 μm; compound III 5 Half inhibition concentrations of HCT-116 cells, MCF-7 cells, 786O cells and HepG2 cells are 0.44 mu M, 0.27 mu M, 0.65 mu M and 0.45 mu M respectively, and inhibition effects are superior to those of positive control drugs Sunitinib and 5-fluorouracil (5-FU), which indicate that the derivative taking pyrrolopyridine as a framework provided by the application has stronger inhibition effect on tumor cells and achieves nanomolar level as a whole.
2. HTRF KinEASE kit for investigating inhibition of PAK5 kinase activity by compounds
The method comprises the following two steps: the first step is an enzymatic reaction step where the PAK5 kinase substrate-2-biotin complex is incubated with the enzyme and ATP is added to initiate the enzymatic reaction. The second step monitors the capture of phosphorylated substrate by the reagent. The final TR-FRET signal is proportional to the phosphorylation level, and Eu is labeled 3+ Antibodies to Cryptate and streptavidin-XL 665 terminate kinase activity. Inhibition = (max-test)/(max-min) ×100% ("max" refers to no compound control, "min" refers to no compound no kinase control).
We evaluated the inhibitory effect of compounds on PAK5 kinase using HTRF kinese kit. The inhibition rate of the compound to PAK5 reaches 50% under the condition of 100nM, and the inhibition rate of the compound III reaches 1 mu M 1 、III 2 、III 3 The inhibition rate of PAK5 reaches more than 80%, and other compounds which are not listed have similar effects. We select the compounds III4 and III5 with better inhibition effect to carry out half inhibition evaluation, and find the compound III 4 、III 5 Half inhibition concentrations for PAK5 reached 14nM and 8nM, respectively, similar to the positive control drug sunitinib (see table 2 for results).
Inhibitory Activity of Compounds of Table 2 on PAK5 kinase
a The experiments were repeated three times at the concentration corresponding to the compound at which kinase activity was inhibited by half, and the results were expressed as Mean ± standard deviation (Mean ± SD).
3. Determination of cytotoxicity of Compounds on Normal cells by CCK-8 method
Normal human tubular epithelial HK-2 cells in logarithmic growth phase in pancreatin digestion dishes, counted by counting plate, and inoculated in 96-well plate. Except that 200 mu L of PBS is added to the peripheral holes, 3000-6000 cells are paved in each hole; setting 4 compound wells, and adding 100 mu L of 10% FBS complete culture medium into each well; after the cells are attached, the culture medium in the 96-well plate is discarded by reverse buckling, the culture medium containing the compounds with different concentrations is added, and the mixture is placed at 37 ℃ and 5 percent CO 2 Culturing in an incubator for 48 hours; after 48 hours, the old culture medium in the 96-well plate is discarded, 90 mu L of serum-free culture medium and 10 mu L of CCK-8 solution are added into each well, and the mixture is uniformly mixed and is operated in a dark place; placing the mixture into an incubator for continuous incubation for 0.5,1,2 and 4 hours; optical Density (OD) measurements at a wavelength of 450nm were read using an microplate reader and repeated three times.
We examined the target compound III by CCK-8 method 5 And III 4 Cytotoxicity against normal human tubular epithelial cells HK-2, the results of which are shown in FIG. 1. At a compound concentration of 1. Mu.M, the viability of the HK-2 cells of groups III4 and III5 was 70.48% and 76.33%, respectively, which is higher than that of 5-FU by 63.75%; when the compound concentration was 5. Mu.M, the cell viability of the III4 and III5 groups was 30%, which was close to 34.95% of the 5-FU group, and the difference was statistically significant, indicating that the compound was better safe. Other compounds of the application also have similar effects.
4. In vivo tumor inhibiting effect of compound under investigation of xenograft tumor nude mouse model
To further evaluate the target compound III 5 Antitumor effects in animals. We used HepG2 cell nude mouse skinIn vivo experiments of the following tumors, the in vivo tumor inhibiting effect of the compound III5 was evaluated. 30 BALB/C nude mice (Male, SPF grade, 18-22 g) were inoculated subcutaneously on the right armpit of each nude mouse 5X 10 6 And (3) establishing a tumor-bearing animal model by using the HepG2 cells. The subcutaneous tumor grows to 100-140mm 3 The nude mice were then randomly divided into 5 groups, namely a control group (Vehicle), a positive control drug sunitinib (20 mg/kg) group and a target compound III 5 High, medium and low concentration groups (50 mg/kg, 35mg/kg and 20 mg/kg). The tumor size and body weight of each nude mouse were measured daily by administering the drug once daily and continuously for 16 days, and the long diameter (a) and short diameter (b) of the tumor of the mouse were measured. Nude mice were sacrificed 16 days after administration by cervical removal, tumor tissues and normal tissues were removed, and hematoxylin-eosin staining (H&E) A. The application relates to a method for producing a fibre-reinforced plastic composite According to the tumor volume calculation formula: v=1/2×a×b 2 Tumor volumes were calculated and tumor inhibition rates were obtained, along with statistical t-test analysis. The results are shown in FIGS. 2-4.
Fig. 2A is a tumor map in nude mice. As shown in the figure, compound III 5 The tumors in the group were significantly reduced compared to the blank. At the same dosage, 20mg/k of Compound III 5 The tumor size of the administration group is equivalent to the treatment effect of the positive control drug sunitinib group. High concentration of Compound III 5 The tumor volume of the (50 mg/kg) administered group was minimal, indicating that a dose-dependent relationship exists.
Fig. 2B is a graph of tumor volume change. As shown, tumor volume in the blank group increased exponentially with time, while tumor volume in the dosing group was significantly smaller than that in the control group. At the same administration concentration, compound III 5 Compared with the positive control medicine, the tumor inhibition rate of the group is improved, and the group is in a dose-dependent relationship.
Fig. 2C is a graph of tumor weight change. As shown, the tumor weights of the dosing groups were reduced to varying degrees from the blank groups. At the same dosing concentration, compound III 5 The average tumor weight of the administration group is reduced by 0.15g compared with that of the positive control drug sunitinib group. With the increase of the dosage, the tumor weight is reduced continuously, and when the dosage is 50mg/kg, the tumor inhibition rate reaches 91.21 percent.
Fig. 3 is a graph showing the change in body weight of each group of tumor-bearing nude mice during the administration period. As shown, there was no significant change in the body weight of the nude mice during the dosing period, indicating that compound III5 had lower toxicity.
Fig. 4 is a HE stained section of normal tissues of nude mice after the end of administration, wherein hematoxylin dye primarily stained the nucleus of tumor cells in bluish violet, while eosin dye primarily stains the cytoplasm and extracellular matrix in red. The results show that each group has a compact cell mass, the cell nucleus is complete and the cell morphology is normal, which indicates that the compound III5 does not cause pathological damage to main organs such as heart, liver, spleen, lung, kidney and the like of nude mice, and has in vivo biosafety.
The above embodiments are only for illustrating the technical solution of the present application, and are not limiting; although the application has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments may be modified or some technical features may be replaced equivalently; such modifications and substitutions do not depart from the spirit of the application.

Claims (4)

1. A compound, or a pharmaceutically acceptable salt thereof, selected from the group consisting of:
2. a pharmaceutical composition comprising a compound of claim 1 or a pharmaceutically acceptable salt thereof as an active ingredient or a primary active ingredient, in combination with a pharmaceutically acceptable carrier.
3. Use of a compound as claimed in claim 1 or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment of a disease associated with the serine/threonine kinase PAK 5.
4. The use according to claim 3, wherein the serine/threonine kinase PAK 5-related disorder is colorectal cancer, liver cancer, gastric cancer, breast cancer, renal cancer or cervical cancer.
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