CN118126040A - Compounds containing tetrahydropyrazolopyridin[3,4-d]pyrimidine structure - Google Patents

Abstract

The present invention relates to a compound containing a tetrahydropyrazolopyridine [3,4-d] pyrimidine structure. Specifically, the compound of the present invention has a structure shown in Formula I, wherein the definitions of each group and substituent are as described in the specification. The present invention also discloses a preparation method of the compound and its use in preventing and/or treating KRAS mutation-related diseases.

Description

Compounds containing tetrahydropyrazolopyridin[3,4-d]pyrimidine structure

Technical Field

The present invention relates to the field of medicine, and in particular to a compound containing a tetrahydropyrazolopyridine [3,4-d] pyrimidine structure.

Background Art

KRAS (Kirsten rat sarcoma viral oncogene homolog) is a rat sarcoma viral oncogene. The KRAS protein encoded by this gene is a small GTPase that participates in the regulation of multiple cell signaling pathways and has an important impact on cell growth, survival, and differentiation. However, mutation of the KRAS gene is a common carcinogenic mutation, which has been shown to occur in 90% of pancreatic cancer, 45% of colorectal cancer, and 35% of lung adenocarcinoma. It can be seen that drug discovery and research targeting KRAS can be said to be a promising direction for the development of anti-tumor drugs. In May 2021, Amgen's Sotorasib was officially approved by the FDA for marketing and is the first small molecule inhibitor targeting KRAS ^G12C . However, rapid adaptive resistance and reactivation of MAPK signals after treatment with KRAS ^G12C inhibitors have been reported, suggesting that this type of inhibitor has drug resistance. Therefore, the development of new KRAS ^G12C- targeted inhibitors and the exploration of new small molecule drugs with new mechanisms of action remain challenging and of great significance.

Using the mechanism of protein targeting chimera (PROteolysis Targeting Chimera, PROTAC) to inhibit the function of KRAS provides a new idea for targeting KRAS. XY-4-88 (Cell Chemical Biology, 2020, 27: 19-31) is the first PROTAC based on KRAS ^G12C inhibitor, but the compound has no degradation activity on endogenous KRAS protein. By changing the ligand of the target protein and the E3 ligase ligand, compound LC-2 (ACS Central Science, 2020, 6: 1367-1375) became the first PROTAC molecule that can degrade endogenous KRAS ^G12C protein, with a DC ₅₀ value of 0.25-0.76 μM. However, as a class of bifunctional molecules, PROTAC has a higher molecular weight (800–1200 Da), more hydrogen bond donors, and a larger polar surface area than typical small molecules, all of which affect its drugability.

Therefore, there is an urgent need in the art to develop a class of small molecule compounds with KRAS protein degradation inhibitory activity.

Summary of the invention

The object of the present invention is to provide a compound represented by formula I and a preparation method thereof and use thereof in preventing and/or treating diseases related to KRAS protein mutation.

The first aspect of the present invention provides a compound, which is a compound represented by Formula I, or a pharmaceutically acceptable salt or solvate thereof,

wherein R ¹ and R ² are each independently selected from the following group: hydrogen, halogen, hydroxyl, amino, cyano, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 3-8 membered heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, substituted or unsubstituted C5-C10 aryl, substituted or unsubstituted 5-10 membered heteroaryl containing 1, 2 or 3 heteroatoms selected from N, O or S The substitutions are each independently substituted by 1, 2, 3, 4, 5 or 6 substituents selected from the group consisting of -(C=O)-NRaRb, -NRaRb, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy-substituted C1-C6 alkyl, C1-C6 alkoxy, oxo (=O), halogen, C3-C8 cycloalkyl, 3-8 membered heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, C5-C10 aryl, 5-10 membered heteroaryl containing 1, 2 or 3 heteroatoms selected from N, O or S;

^R3 is selected from the group consisting of substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl; the substitutions are each independently substituted by 1, 2, 3, 4, 5 or 6 substituents selected from the group consisting of halogen, hydroxy, -NRaRb, 4-6 membered heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S;

R ⁴ is selected from the following group: substituted or unsubstituted C5-C10 aryl, substituted or unsubstituted 5-10 membered heteroaryl containing 1, 2 or 3 heteroatoms selected from N, O or S; the substitutions are each independently substituted by 1, 2, 3, 4, 5 or 6 substituents selected from the following group: halogen, hydroxyl, amino, C1-C6 alkyl, halogenated C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C3-C8 cycloalkyl;

Ra and Rb are each independently selected from the following groups: hydrogen, C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl, 3-8 membered heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, C5-C10 aryl, 5-10 membered heteroaryl containing 1, 2 or 3 heteroatoms selected from N, O or S.

In another preferred embodiment, R ¹ and R ² are each independently selected from the following group: hydrogen, halogen, hydroxyl, amino, cyano, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 3-8 membered heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, substituted or unsubstituted C5-C1 0 aryl, substituted or unsubstituted 5-10 membered heteroaryl containing 1, 2 or 3 heteroatoms selected from N, O or S; the substitutions are each independently substituted by 1, 2, 3, 4, 5 or 6 substituents selected from the group consisting of: -(C=O)-NRaRb, -NRaRb, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy substituted C1-C6 alkyl, C1-C6 alkoxy, oxo (=O), halogen;

Ra and Rb are as defined in claim 1.

In another preferred embodiment, R ¹ and R ² are each independently selected from the following group: hydrogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 3-8 membered heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, substituted or unsubstituted 5-10 membered heteroaryl containing 1, 2 or 3 heteroatoms selected from N, O or S; the substitutions each independently refer to substitutions by 1, 2, 3, 4, 5 or 6 substituents selected from the following group: -(C＝O)-NRaRb, -NRaRb, C1-C6 alkyl, C1-C6 alkyl substituted with C1-C6 alkoxy, oxo (＝O), halogen.

In another preferred embodiment, R ¹ is selected from the following groups: substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 3-8 membered heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S; the substitutions each independently refer to substitutions by 1, 2, 3, 4, 5 or 6 substituents selected from the following group: -(C＝O)-NRaRb, -NRaRb, C1-C6 alkyl, C1-C6 alkoxy substituted C1-C6 alkyl, oxo (＝O), halogen.

In another preferred embodiment, ^R2 is selected from the following groups: hydrogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted 3-8 membered heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, substituted or unsubstituted 5-7 membered heteroaryl containing 1, 2 or 3 heteroatoms selected from N, O or S; the substitutions each independently refer to substitutions by 1, 2, 3, 4, 5 or 6 substituents selected from the following group: -(C＝O)-NRaRb, -NRaRb, C1-C6 alkyl, C1-C6 alkoxy substituted C1-C6 alkyl, oxo (＝O), halogen.

In another preferred embodiment, ^R2 is selected from the following groups: hydrogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted 3-8 membered heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, substituted or unsubstituted 8-10 membered heteroaryl containing 1, 2 or 3 heteroatoms selected from N, O or S; the substitutions each independently refer to substitutions by 1, 2, 3, 4, 5 or 6 substituents selected from the following group: -(C＝O)-NRaRb, -NRaRb, C1-C6 alkyl, C1-C6 alkoxy substituted C1-C6 alkyl, oxo (＝O), halogen.

In another preferred embodiment, R ³ is selected from the following group: substituted or unsubstituted C2-C4 alkenyl, substituted or unsubstituted C2-C4 alkynyl; the substitutions are each independently substituted by 1, 2, 3, 4, 5 or 6 substituents selected from the following group: halogen, hydroxyl, -NRaRb;

Ra and Rb are as defined in claim 1.

In another preferred embodiment, R ⁴ is selected from the following groups: substituted or unsubstituted C8-C10 aryl, substituted or unsubstituted 8-10 membered heteroaryl containing 1, 2 or 3 heteroatoms selected from N, O or S; the substitutions each independently refer to substitutions by 1, 2, 3, 4, 5 or 6 substituents selected from the following group: halogen, hydroxyl, amino, C1-C6 alkyl, halogenated C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C3-C8 cycloalkyl.

In another preferred embodiment, ^R4 is selected from the following groups: substituted or unsubstituted C8-C10 aryl, substituted or unsubstituted 8-10 membered heteroaryl containing 1, 2 or 3 heteroatoms selected from N, O or S; the substitutions each independently refer to substitutions by 1, 2, 3, 4, 5 or 6 substituents selected from the following group: halogen, hydroxyl, amino, C1-C6 alkyl.

In another preferred embodiment, Ra and Rb are each independently selected from the following group: hydrogen, C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl.

In another preferred embodiment, Ra and Rb are each independently selected from the following group: hydrogen, C1-C3 alkyl.

In another preferred embodiment, the compound is selected from the following group:

The second aspect of the present invention provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a safe and effective amount of one or more compounds described in the first aspect of the present invention.

The third aspect of the present invention provides a use of the compound according to the first aspect of the present invention for preparing a medicament for preventing and/or treating a KRAS mutation-related disease.

In another preferred embodiment, the KRAS mutation is selected from the following group: KRAS ^G12C mutation and KRAS ^G12D mutation.

In another preferred embodiment, the KRAS mutation is a KRAS ^G12C mutation.

In another preferred embodiment, the KRAS mutation-related disease is a KRAS ^G12C mutation-related disease.

In another preferred embodiment, the KRAS mutation-related disease is cancer.

In another preferred embodiment, the KRAS mutation-related disease is selected from the following group: pancreatic cancer, colorectal cancer, lung adenocarcinoma, lung cancer, bile duct cancer, cervical cancer, bladder cancer, liver cancer, breast cancer, endometrial cancer, skin cancer, ovarian cancer, gastric cancer, urinary tract cancer, soft tissue sarcoma, esophageal cancer, multiple myeloma, prostate cancer, kidney cancer, gastrointestinal neuroendocrine tumors, gastrointestinal stromal tumors, head and neck cancer, glioma, salivary gland cancer, bone cancer, anal cancer, thyroid cancer, melanoma, mature B cell tumors, small intestine cancer, and ampullary cancer.

It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here.

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1 shows the degradation effect of compound S9 on KRAS protein in MIA PaCa-2 and its effect on phosphorylation of downstream effector proteins.

FIG2 is a graph showing the growth inhibition rate of compound S9 on MIA PaCa-2 cells, with IC ₅₀ =0.34±0.04 μM.

FIG3 is a curve diagram showing the in vivo tumor volume inhibition effect of compound S9 in a nude mouse xenograft tumor model of pancreatic cancer MIA PaCa-2 cells.

DETAILED DESCRIPTION

After long-term and in-depth research, the inventors have obtained a class of compounds with excellent KRAS mutation (especially KRAS ^G12C mutation) inhibition and/or degradation activity through structural optimization, and the compounds have excellent inhibitory activity and selective degradation activity for KRAS ^G12C mutant protein, and the compounds are expected to be developed as new small molecule inhibitors and degraders of KRAS ^G12C . On this basis, the inventors have completed the present invention.

the term

In the present invention, unless otherwise specified, the terms used have the general meanings well known to those skilled in the art.

In the present invention, the term "halogen" refers to F, Cl, Br or I.

In the present invention, "C1-C6 alkyl" refers to a straight or branched alkyl group comprising 1 to 6 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, neopentyl, tert-pentyl, or the like.

In the present invention, the term "C2-C6 alkenyl" refers to a straight or branched alkenyl group having 2 to 6 carbon atoms and containing one double bond, including but not limited to ethenyl, propenyl, butenyl, isobutenyl, pentenyl and hexenyl.

In the present invention, the term "C2-C6 alkynyl" refers to a straight or branched alkynyl group having 2 to 6 carbon atoms and containing one triple bond, including but not limited to ethynyl, propynyl, butynyl, isobutynyl, pentynyl and hexynyl.

In the present invention, the term "C3-C8 cycloalkyl" refers to a cyclic alkyl group having 3 to 8 carbon atoms in the ring, including but not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl and the like.

In the present invention, the term "C1-C6 alkoxy" refers to a straight or branched alkoxy group having 1 to 6 carbon atoms, including but not limited to methoxy, ethoxy, propoxy, isopropoxy and butoxy, etc. Preferably, it is C1-C4 alkoxy.

In the present invention, the term "heterocycloalkyl" refers to a 4-8 membered heterocycloalkyl group containing 1, 2 or 3 heteroatoms selected from N, O and S, including (but not limited to) the following groups:

In the present invention, the term "aromatic ring" or "aryl group" has the same meaning, preferably "C6-C10 aryl group". The term "C6-C10 aryl group" refers to an aromatic ring group having 6 to 10 carbon atoms without heteroatoms in the ring, such as phenyl, naphthyl, etc.

In the present invention, the term "aromatic heterocycle" or "heteroaryl" has the same meaning and refers to a heteroaromatic group containing one to multiple heteroatoms. For example, "C3-C10 heteroaryl" refers to an aromatic heterocycle containing 1 to 4 heteroatoms selected from oxygen, sulfur and nitrogen and 3 to 10 carbon atoms. Non-limiting examples include: furanyl, thienyl, pyridyl, pyrazolyl, pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, imidazolyl, tetrazolyl, etc. The heteroaryl ring can be fused to an aryl, heterocyclic or cycloalkyl ring, wherein the ring connected to the parent structure is a heteroaryl ring. The heteroaryl group can be optionally substituted or unsubstituted.

In the present invention, the term "halo" means substituted with halogen.

In the present invention, the term "substituted" refers to one or more hydrogen atoms on a specific group being replaced by a specific substituent. The specific substituent is the substituent described above, or the substituent appearing in each embodiment. Unless otherwise specified, a substituted group may have a substituent selected from a specific group at any substitutable position of the group, and the substituent may be the same or different at each position. It should be understood by those skilled in the art that the combination of substituents contemplated by the present invention is those stable or chemically feasible combinations. The substituents include, for example (but not limited to): halogen, hydroxyl, carboxyl (-COOH), C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkyl, 3- to 12-membered heterocyclic radical, aryl, heteroaryl, C1-C8 aldehyde, C2-C10 acyl, C2-C10 ester, amino, C1-C6 alkoxy, C1-C10 sulfonyl, etc.

In the present invention, the term 1-6 refers to 1, 2, 3, 4, 5 or 6. Other similar terms independently have similar meanings. The term "plurality" refers to 2-6, such as 2, 3, 4, 5 or 6.

It should be understood that when a group is present in multiple different positions of a compound at the same time, its definition at each position is independent of each other and may be the same or different. That is, the term "selected from the following group:" has the same meaning as the term "each independently selected from the following group:".

Compound

The present invention provides a class of compounds containing a 5,6,7,8-tetrahydropyrazolo[1,5-a]pyridine[3,4-d]pyrimidine structure, wherein the compounds exhibit degradation activity (DC ₅₀ =0.1 μM) on KRAS protein in KRAS ^G12C mutant cell lines. Compared with KRAS degraders reported in existing literature, the compounds with KRAS degradation function provided by the present invention do not need to connect the KRAS inhibitor to other functional molecules through a lengthy connecting chain, thereby having a smaller molecular weight (<700 Da) and more potential drugability, and can inhibit the proliferation and growth of KRAS ^G12C mutant cells.

The present invention provides a compound, which is a compound represented by Formula I, or a pharmaceutically acceptable salt or solvate thereof,

wherein each group is as defined above.

In another preferred embodiment, in the compound, any one of R ¹ , R ² , R ³ , R ⁴ , Ra, and Rb is independently a corresponding group in the specific compound described in the present application.

As used herein, the term "pharmaceutically acceptable salt" refers to a salt formed by a compound of the present invention and an acid or base that is suitable for use as a drug. Pharmaceutically acceptable salts include inorganic salts and organic salts. A preferred class of salts is a salt formed by a compound of the present invention and an acid. Suitable acids for forming salts include, but are not limited to, inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, and phosphoric acid; organic acids such as formic acid, acetic acid, trifluoroacetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, benzoic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, and naphthalenesulfonic acid; and amino acids such as proline, phenylalanine, aspartic acid, and glutamic acid.

Another preferred salt is a salt of the compound of the present invention and a base, such as an alkali metal salt (e.g., sodium salt or potassium salt), an alkaline earth metal salt (e.g., magnesium salt or calcium salt), an ammonium salt (e.g., lower alkanolammonium salt and other pharmaceutically acceptable amine salts), for example, methylamine salt, ethylamine salt, propylamine salt, dimethylamine salt, trimethylamine salt, diethylamine salt, triethylamine salt, tert-butylamine salt, ethylenediamine salt, hydroxyethylamine salt, dihydroxyethylamine salt, trihydroxyethylamine salt, and amine salts formed from morpholine, piperazine, and lysine, respectively.

The term "solvate" refers to a complex in which the compound of the present invention is coordinated with solvent molecules to form a specific ratio.

It should be understood that the compounds of the present invention can be prepared by the methods shown in the embodiments of the present invention or by methods well known to those skilled in the art, and the specific methods shown in the following examples do not constitute any limitation to the present invention. The compounds of the present invention can also be conveniently prepared by optionally combining various synthetic methods described in this specification or known in the art, and such combinations can be easily performed by those skilled in the art to which the present invention belongs.

Typically, the raw materials and reagents used in the process for preparing the compounds of the present invention can be purchased through commercial channels unless otherwise specified.

Pharmaceutical compositions and methods of administration

The present invention also provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and one or more safe and effective amounts of the compounds.

Since the compounds of the present invention have excellent anti-tumor activity, the compounds of the present invention and their various crystal forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and pharmaceutical compositions containing the compounds of the present invention as the main active ingredient can be used to treat, prevent and alleviate tumor-related diseases.

The pharmaceutical composition of the present invention comprises a safe and effective amount of the compound of the present invention or a pharmacologically acceptable salt thereof and a pharmacologically acceptable excipient or carrier. Wherein "safe and effective amount" means: the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. Usually, the pharmaceutical composition contains 1-2000 mg of the compound of the present invention per dose, and more preferably, contains 10-1000 mg of the compound of the present invention per dose. Preferably, the "one dose" is a capsule or tablet.

"Pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with the compounds of the present invention and with each other without significantly reducing the efficacy of the compounds. Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as ), wetting agents (such as sodium lauryl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

The pharmaceutical composition is in the form of injection, capsule, tablet, pill, powder or granule.

The administration method of the compound or pharmaceutical composition of the present invention is not particularly limited, and representative administration methods include (but are not limited to): oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration.

Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and acacia; (c) humectants, for example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

Solid dosage forms such as tablets, pills, capsules, pills and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifiers, and the release of the active compound or compounds in such compositions may be delayed in a certain part of the digestive tract. Examples of embedding components that can be used are polymeric substances and waxes. If necessary, the active compound can also be formed into microencapsulated form with one or more of the above-mentioned excipients.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide and oils, in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances.

Besides such inert diluents, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methanol and agar, or mixtures of these substances, and the like.

Compositions for parenteral injection may include physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

Dosage forms for topical administration of the compounds of the invention include ointments, powders, patches, sprays and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be required.

The compounds of the present invention can be administered alone or in combination with other pharmaceutically acceptable compounds (such as anti-tumor drugs).

The treatment method of the present invention can be used alone or in combination with other treatment methods or therapeutic drugs.

When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the dosage during administration is a pharmaceutically effective dosage, and for a person weighing 60 kg, the daily dosage is usually 1 to 2000 mg, preferably 50 to 1000 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the health status of the patient, which are all within the skill of a skilled physician.

Compared with the prior art, the present invention has the following main advantages:

(1) The compound has excellent KRAS degradation inhibitory activity;

(2) The compound has excellent degradation inhibitory activity against KRAS protein;

(3) The compound has the following advantages: excellent drugability, excellent pharmacokinetic properties, easy preparation, and excellent safety;

(4) The compound is a KRAS degrader and inhibits KRAS activity;

(5) The compound exhibits in vivo tumor inhibitory activity against the KRAS ^G12C mutation transplant tumor model.

The present invention is further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples that do not specify specific conditions are usually based on conventional conditions such as Sambrook et al., Molecular Cloning: Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989) or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight.

Unless otherwise defined, all professional and scientific terms used herein have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein can be applied to the methods of the present invention. The preferred implementation methods and materials described herein are for demonstration purposes only.

Preparation Example 1 Preparation of Compound S1:

Synthesis of compound 1-3:

Compound 1-1 (5.0 g, 40.6 mmol) and 1-2 (12.1 g, 40.6 mmol) were dissolved in 20 ml of acetic acid and stirred at 100°C for 1 hour. After cooling to room temperature, most of the solvent was removed under reduced pressure, neutralized with saturated sodium bicarbonate aqueous solution, extracted with dichloromethane, and the organic phase was dried over anhydrous sodium sulfate. The mixture was subjected to column chromatography to obtain white solid compound 1-3.

Synthesis of compound 1-4:

Compound 1-3 (8.0 g, 25.0 mmol) was suspended in 20 ml of toluene, phosphorus oxychloride (23.2 mL, 250 mmol) and N, N-diisopropylethylamine (16.6 mL, 100 mmol) were slowly added to the reaction under an ice bath, and then the reaction was stirred at 100°C for 1 hour. After cooling to room temperature, most of the solvent was removed by evaporation under reduced pressure, and the mixture was slowly diluted with saturated sodium bicarbonate aqueous solution, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate. The mixture was subjected to column chromatography to obtain yellow solid compound 1-4.

Synthesis of compound 1-6:

Compound 1-4 (3.0 mg, 8.85 mmol) and compound 1-5 (2.39 g, 10.6 mmol) were dissolved in 10 ml of dimethyl sulfoxide, and N, N-diisopropylethylamine (4.40 mL, 26.6 mmol) was slowly added at room temperature. The reaction was then stirred at 80°C overnight. The mixture was cooled to room temperature, diluted with saturated brine, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate and subjected to column chromatography to obtain yellow foam solid compound 1-6.

Synthesis of compound 1-7:

Compound 1-6 (1.0 g, 1.90 mmol) was dissolved in 15 ml of methanol, 5% palladium carbon (403 mg, 0.190 mmol) and ammonium formate (1.19 g, 19.0 mmol) were added, and the mixture was heated under reflux overnight. The mixture was filtered through celite, and the filtrate was dried and subjected to column chromatography to obtain yellow solid compound 1-7.

Synthesis of compound 1-9:

Compound 1-7 (428 mg, 0.978 mmol) and compound 1-8 (429 mg, 1.27 mmol) were dissolved in 10 ml of toluene, tri(dibenzylideneacetone)dipalladium (179 mg, 0.196 mmol), 2-dicyclohexylphosphine-2',6'-diisopropoxy-1,1'-biphenyl (183 mg, 0.391 mmol) and cesium carbonate (956 mg, 2.93 mmol) were added, and the mixture was reacted at 100°C for 5 hours after nitrogen substitution. The mixture was cooled to room temperature, diluted with water, extracted with ethyl acetate, and the organic phase was combined and dried over anhydrous sodium sulfate, and the mixture was subjected to column chromatography to obtain white solid compound 1-9.

Synthesis of compound S1:

Compound 1-9 (132 mg, 0.19 mmol) was dissolved in 2 ml of dichloromethane, and 2 ml of trifluoroacetic acid was slowly added under ice bath. After the reaction solution was placed at room temperature for 1 hour, the reaction solvent was removed by vacuum concentration. Then, 3 ml of dichloromethane was added to the mixture, N, N-diisopropylethylamine (95 μL, 0.57 mmol) was added to the reaction system under ice bath, and acryloyl chloride (19 mg, 0.21 mmol) was slowly added dropwise under stirring. The reaction solution was stirred at 0°C for 30 minutes, diluted with saturated sodium bicarbonate aqueous solution, extracted with dichloromethane, and the organic phases were combined and dried over anhydrous sodium sulfate. After the solvent was dried under reduced pressure, the mixture was separated by column chromatography to obtain a yellow solid compound. ¹ H NMR (400 MHz, Methanol-d ₄ )δ8.03(d,J＝8.4Hz,1H),7.59(d,J＝8.2Hz,1H),7.37–7.28(m,1H),7.26–7.17(m,1H),6.97–6.71(m,3H),6.28(d,J＝17.4Hz,1H),6.16(s,1H),5.82(d ,J＝11.7Hz,1H),5.32–5.09(m, 0.5H),4.83–4.47(m,1H),4.37–4.15(m,2H),4.13–4.03(m,0.5H),3.94–3.51(m,4H),3.50–3.29(m,4H),3.19–2.89(m,3H),2.17–2.06(m,1H),1. 10–1.00(m,2H),0.93–0.83(m,2H).

Preparation Example 2 Preparation of Compound S2:

Synthesis of compound 2-1:

Compound 1-6 (3.0 g, 5.69 mmol) was dissolved in 15 ml of N,N-dimethylformamide and replaced with nitrogen. N-bromosuccinimide (1.11 g, 6.25 mmol) was slowly added in batches under an ice bath and the reaction was continued at this temperature for 1 hour. Saturated sodium bicarbonate solution was added to the reaction solution, and ethyl acetate was extracted. The organic phase was washed with saturated brine and dried over anhydrous sodium sulfate, then concentrated and purified by column chromatography to obtain a yellow solid 2-1.

Synthesis of compound 2-2:

Compound 2-1 (1.0 g, 1.65 mmol) and N,N-dimethylacrylamide (327 mg, 3.30 mmol) were dissolved in 10 ml of acetonitrile, and N,N-diisopropylethylamine (2.73 mL, 16.5 mmol) and tris(dibenzylideneacetone)dipalladium (151 mg, 0.165 mmol) were added, and the atmosphere was replaced with nitrogen and reacted overnight at 80°C. The reaction solution was cooled to room temperature, diluted with water and extracted with ethyl acetate, and the organic phase was washed with saturated brine and dried over anhydrous sodium sulfate, then concentrated and subjected to column chromatography to obtain a light yellow oil 2-2.

Synthesis of compound 2-3:

Compound 2-2 (500 mg, 0.800 mmol) was dissolved in 10 ml of methanol, 5% palladium carbon (170 mg, 0.080 mmol) and ammonium formate (505 mg, 8.00 mmol) were added, and the mixture was heated under reflux overnight. The mixture was filtered through celite, the filtrate was dried by rotary evaporation, and yellow solid compound 2-3 was obtained by column chromatography.

Synthesis of compound 2-4:

Compound 2-3 (224 mg, 0.417 mmol) and compound 1-8 (183 mg, 0.543 mmol) were dissolved in 5 ml of toluene, tri(dibenzylideneacetone)dipalladium (76 mg, 0.083 mmol), 2-dicyclohexylphosphine-2',6'-diisopropoxy-1,1'-biphenyl (78 mg, 0.167 mmol) and cesium carbonate (408 mg, 1.25 mmol) were added, and the mixture was reacted at 100°C for 5 hours after nitrogen substitution. The mixture was cooled to room temperature, diluted with water, extracted with ethyl acetate, and the organic phase was combined and dried over anhydrous sodium sulfate, and then column chromatography was performed to obtain a white solid compound 2-4.

Synthesis of compound S2:

Compound 1-9 (87 mg, 0.110 mmol) was dissolved in 1.5 ml of dichloromethane, and 1.5 ml of trifluoroacetic acid was slowly added under ice bath. After the reaction solution was placed at room temperature for 1 hour, the reaction solvent was removed by vacuum concentration. Then, 3 ml of dichloromethane was added to the mixture, and N, N-diisopropylethylamine (55 μL, 0.330 mmol) was added to the reaction system under ice bath, and acryloyl chloride (11 mg, 0.121 mmol) was slowly added dropwise under stirring. The reaction solution was stirred at 0°C for 30 minutes, diluted with saturated sodium bicarbonate aqueous solution, extracted with dichloromethane, and the organic phases were combined and dried over anhydrous sodium sulfate. After the solvent was dried under reduced pressure, the mixture was separated by column chromatography to obtain a yellow solid compound. ¹ H NMR (400 MHz, Methanol-d ₄ )δ8.07(d,J＝8.4Hz,1H),7.63(d,J＝8.2Hz,1H),7.43–7.32(m,1H),7.31–7.20(m,1H),7.02–6.72(m,3H),6.38–6.24(m,1H),5.93–5.79(m,1H),5.21 (s,0.5H),4.59 (s,1H),4.42–4.24(m,2H),4.23–4.03(m,0.5H),4.01–3.32(m,8H),3.30–3.08(m,3H),3.08–2.80(m,8H),2.81–2.65(m,2H),2.21–2.08(m,1H),1 .11–0.97(m,4H).

Preparation Example 3 Preparation of Compound S3:

Synthesis of compound 3-2:

Compound 2-1 (1.0 g, 1.65 mmol) and 3-1 (849 mg, 2.47 mmol) were dissolved in 10 ml of 1,4-dioxane, and 2.5 ml of water was added, followed by tetrakis(triphenylphosphine)palladium (191 mg, 0.165 mmol) and sodium carbonate (524 mg, 4.95 mmol), replaced with nitrogen, and reacted overnight at 100°C. After the reaction solution was cooled to room temperature, water and ethyl acetate were added, filtered through diatomaceous earth, and then extracted with ethyl acetate. The organic phase was washed with saturated brine and dried over anhydrous sodium sulfate, then concentrated and eluted by column chromatography to obtain a yellow solid 3-2.

Synthesis of compound 3-3:

Compound 3-2 (631 mg, 0.849 mmol) and N, N-diisopropylethylamine (0.422 mL, 2.55 mmol) were dissolved in 8 ml of 1,2-dichloroethane, and 1-chloroethyl chloroformate (0.229 mL, 2.12 mmol) was slowly added at 0°C. The reaction was then continued to stir at 0°C for one hour, the solvent was dried under reduced pressure, the residue was dissolved in 10 ml of methanol, and the reaction was carried out at 60°C for one hour. The mixture was cooled to room temperature, diluted with saturated sodium bicarbonate, and extracted with ethyl acetate. The organic phases were combined, and the mixture was dried over anhydrous sodium sulfate and subjected to column chromatography to obtain a white solid compound 3-3.

Synthesis of compound 3-4:

Compound 3-3 (289 mg, 0.443 mmol) and compound 1-8 (194 mg, 0.576 mmol) were dissolved in 5 ml of dry toluene, tri(dibenzylideneacetone)dipalladium (81 mg, 0.086 mmol), 2-dicyclohexylphosphine-2',6'-diisopropoxy-1,1'-biphenyl (83 mg, 0.177 mmol) and cesium carbonate (433 mg, 1.33 mmol) were added under nitrogen protection and reacted at 100°C for 5 hours. Cooled to room temperature, diluted with water, extracted with ethyl acetate. Combined organic phases, dried over anhydrous sodium sulfate, and the mixture was subjected to column chromatography to obtain yellow solid compound 3-4.

Synthesis of compound 3-5:

Compound 3-4 (173 mg, 0.190 mmol) was dissolved in 2 ml of methanol, and 41 mg of 5% palladium carbon catalyst and ammonium formate (120 mg, 1.90 mmol) were added, and the mixture was heated to reflux overnight. The metal palladium was filtered off with diatomaceous earth, and the filtrate was dried by rotary evaporation, dissolved in methanol, and a formaldehyde aqueous solution (71 μL, 37% in H ₂ O, 0.950 mmol) and a drop of formic acid were added. Sodium cyanoborohydride (36 mg, 0.571 mmol) was slowly added at 0°C and reacted for one hour. Diluted with saturated sodium bicarbonate, extracted with ethyl acetate (3×10 mL). The organic phases were combined, and the mixture was dried over anhydrous sodium sulfate and subjected to column chromatography to obtain white solid compound 3-5.

Synthesis of compound S3:

Compound 3-5 (104 mg, 0.131 mmol) was dissolved in 2 ml of dichloromethane, and 2 ml of trifluoroacetic acid was slowly added under stirring. After the reaction solution was left to react at room temperature for 1 hour, the reaction solvent was removed by vacuum concentration. Then, 3 ml of dichloromethane was added to the mixture, the reaction system was placed at 0°C and N, N-diisopropylethylamine (65 μL, 0.394 mmol) was added, and acryloyl chloride (13 mg, 0.145 mmol) was slowly added dropwise under stirring. The reaction solution was stirred at 0°C for 30 minutes, diluted with saturated sodium bicarbonate aqueous solution, and extracted with dichloromethane. The organic phases were combined, dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure. The mixture was separated by column chromatography to obtain a white solid compound S3. ¹ H NMR (400 MHz, Methanol-d ₄ )δ8.04(d,J＝8.4Hz,1H),7.61(d,J＝8.2Hz,1H),7.35(dd,J＝8.1,6.6Hz,1H),7.28–7.19(m,1H),6.95–6.75(m,3H),6.28(d,J＝16.7Hz,1H),5.83(d,J＝10 .4Hz,1H),5.26–5.13(m,0. 5H),4.76–4.54(m,2H),4.38–4.19(m,2H),4.15–4.02(m,0.5H),3.96–3.33(m,8H),3.22–2.74(m,7H),2.50–2.38(m,2H),2.36–2.11(m,5H),1.8 0–1.76(m,1H),1.08–0.97(m,4H).

Preparation Example 4 Preparation of Compound S4:

Synthesis of compound 4-2:

Compound 2-1 (2.0 g, 3.30 mmol) and 4-1 (1.16 g, 4.95 mmol) were dissolved in 20 ml of 1,4-dioxane, 5 ml of water was added, and then tetrakis(triphenylphosphine)palladium (381 mg, 0.330 mmol) and sodium carbonate (1.05 g, 9.89 mmol) were added, and nitrogen was replaced, and the reaction was carried out overnight at 100°C. After the reaction solution was cooled to room temperature, water and ethyl acetate were added, filtered with celite, and then extracted with ethyl acetate. The organic phase was washed with saturated brine and dried over anhydrous sodium sulfate, and then concentrated and eluted with column chromatography to obtain a yellow solid 4-2.

Synthesis of compound 4-3:

Compound 4-2 (500 mg, 0.788 mmol) was dissolved in 2 ml of methanol, and 168 mg of 5% palladium carbon catalyst and ammonium formate (497 mg, 7.88 mmol) were added, and the mixture was heated under reflux overnight. The metal palladium was filtered off with diatomaceous earth, and the filtrate was dried with anhydrous sodium sulfate, and the mixture was dried with column chromatography to obtain a white solid compound 4-3.

Synthesis of compound 4-4:

Compound 4-3 (324 mg, 0.595 mmol) and compound 1-8 (260 mg, 0.773 mmol) were dissolved in 8 ml of dry toluene, tri(dibenzylideneacetone)dipalladium (109 mg, 0.119 mmol), 2-dicyclohexylphosphine-2',6'-diisopropoxy-1,1'-biphenyl (111 mg, 0.238 mmol) and cesium carbonate (581 mg, 1.78 mmol) were added under nitrogen protection and reacted at 100°C for 5 hours. The mixture was cooled to room temperature, diluted with water, and extracted with ethyl acetate. The organic phases were combined, and the mixture was dried over anhydrous sodium sulfate and subjected to column chromatography to obtain yellow solid compound 4-4.

Synthesis of compound S4:

Compound 4-4 (89 mg, 0.107 mmol) was dissolved in 1 ml of dichloromethane, and 1 ml of trifluoroacetic acid was slowly added under stirring. After the reaction solution was left to react at room temperature for 1 hour, the reaction solvent was removed by vacuum concentration. Then, 2 ml of dichloromethane was added to the mixture, the reaction system was placed at 0°C and N, N-diisopropylethylamine (53 μL, 0.320 mmol) was added, and acryloyl chloride (10 mg, 0.108 mmol) was slowly added dropwise under stirring. The reaction solution was stirred at 0°C for 30 minutes, diluted with saturated sodium bicarbonate aqueous solution, and extracted with dichloromethane. The organic phases were combined, dried over anhydrous sodium sulfate, and after removing the solvent under reduced pressure, the mixture was separated by column chromatography to obtain a white solid compound S4. ¹ H NMR (400 MHz, Methanol-d ₄ )δ8.07–7.92(m,3H),7.62–7.53(m,1H),7.35–7.29(m,1H),7.26–7.17(m,1H),6.96–6.72(m,3H),6.62(dd,J＝8.7,6.2Hz,1H),6.29(d,J＝16.8Hz,1H) ,5.88–5.74(m ,1H),5.26–5.11(m,0.5H),4.79–4.51(m,1H),4.40–4.17(m,2H),4.16–4.00(m,0.5H),3.94–3.32(m,10H),3.26–2.78(m,4H),2.17–2.06(m,1H) ,1.14–1.02(m,4H).

Preparation Example 5 Preparation of Compound S5:

The substrate was replaced and the compound S5 was synthesized by referring to the compound Example S4. ¹ H NMR (400 MHz, Methanol-d ₄ )δ8.11–8.00(m,3H),7.86–7.79(m,1H),7.69–7.58(m,2H),7.35(t,J＝7.4Hz,1H),7.28–7.21(m,1H),7.01–6.72(m,3H),6.31(d,J＝16.7Hz,1H),5.9 2–5.81(m,1H),5.22(s,0.5H),4.81–4.22(m,5H),4.19–4.03(m,0.5H),3.97–3.37(m,9H),3.29–2.83(m,7H),2.30–2.18(m,1H),1.16–1.07(m,4 H).

Preparation Example 6 Preparation of Compound S6:

The substrate was replaced and the compound S6 was synthesized by referring to the compound Example S4. ¹ H NMR (400 MHz, Methanol-d ₄ )δ8.11–8.00(m,2H),7.86–7.79(m,1H),7.70–7.58(m,1H),7.37(d,J＝7.4Hz,1H),7.28–7.18(m,1H),7.03–6.75(m,3H),6.31(d,J＝16.7Hz,1H),5.9 1–5.80(m,1H),5.23–5.11(s,0.5H),4.83–4.28(m,3H),4.16–4.03(m,0.5H),3.95–3.39(m,9H),3.27–2.79(m,5H),2.15–2.03(m,1H),1.09–1.0 1(m,4H).

Preparation Example 7 Preparation of Compound S7:

Synthesis of compound 7-2:

Take acetonitrile (2.62mL, 50.1mmol) and dissolve it in 50ml of anhydrous tetrahydrofuran. Under nitrogen protection, place the reaction at -78℃ and slowly add bis(trimethylsilyl) lithium amide (33.4mL, 1.0M in THF, 33.4mmol). After stirring for 10 minutes at this temperature, add compound 7-1 (5.00g, 16.7mmol) in small batches. After continuing to react for one hour, move the reaction solution to room temperature. After quenching with saturated ammonium chloride aqueous solution, extract with ethyl acetate, dry the organic phase with anhydrous sodium sulfate and concentrate under reduced pressure to remove the solvent. Then, the mixture obtained by spin drying is dissolved in 50mL of ethanol, and hydrazine acetate (2.31g, 25.1mmol) is added and reacted at 50℃ overnight. Then, dilute with aqueous solution and extract with ethyl acetate. Combine the organic phases, dry the mixture with anhydrous sodium sulfate, and obtain yellow foam solid compound 7-2 by column chromatography.

Synthesis of compound 7-3:

Compound 7-2 (3.4 g, 10.6 mmol) and N-benzyl-3-oxopiperidine-4-carboxylic acid ethyl ester hydrochloride (1-2, 3.14 g, 10.6 mmol) were dissolved in 30 ml of glacial acetic acid and stirred at 100°C for one hour. After cooling to room temperature and removing most of the solvent under reduced pressure, the mixture was neutralized with saturated sodium bicarbonate aqueous solution, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate. The mixture was subjected to column chromatography to obtain yellow foam solid compound 7-3.

Synthesis of compound 7-4:

Compound 7-3 (3.0 g, 5.77 mmol) was suspended in 20 ml of toluene, phosphorus oxychloride (1.66 mL, 17.3 mmol) and N, N-diisopropylethylamine (3.83 mL, 23.1 mmol) were slowly added to the reaction under ice bath, and then the reaction was stirred at 100 ° C for 1 hour. Cooled to room temperature, most of the solvent was evaporated under reduced pressure, slowly diluted with saturated sodium bicarbonate aqueous solution, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate. The mixture was subjected to column chromatography to obtain yellow solid compound 7-4.

Synthesis of compound 7-5:

Compound 7-4 (1.67 g, 3.10 mmol) and compound 1-5 (839 mg, 3.72 mmol) were dissolved in 15 ml of dimethyl sulfoxide, and N, N-diisopropylethylamine (1.54 mL, 9.31 mmol) was slowly added at room temperature. The reaction was then stirred at 80°C overnight. The mixture was cooled to room temperature, diluted with saturated brine, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate and subjected to column chromatography to obtain yellow foam solid compound 7-5.

Synthesis of compound 7-6:

Compound 3-2 (1.35 g, 1.86 mmol) and N, N-diisopropylethylamine (0.923 mL, 5.57 mmol) were dissolved in 15 mL of 1,2-dichloroethane, and 1-chloroethyl chloroformate (0.501 mL, 4.64 mmol) was slowly added at 0°C. The reaction was then continued to stir at 0°C for one hour, the solvent was dried under reduced pressure, the residue was dissolved in 20 mL of methanol, and the reaction was carried out at 60°C for one hour. The mixture was cooled to room temperature, diluted with saturated sodium bicarbonate, and extracted with ethyl acetate. The organic phases were combined, and the mixture was dried over anhydrous sodium sulfate and subjected to column chromatography to obtain white solid compound 7-6.

Synthesis of compound 7-7:

Compound 7-6 (452 mg, 0.710 mmol) and 1-bromo-8-methylnaphthalene (204 mg, 0.923 mmol) were dissolved in 10 ml of dry toluene, and tris(dibenzylideneacetone)dipalladium (130 mg, 0.142 mmol), 2-dicyclohexylphosphine-2',6'-diisopropoxy-1,1'-biphenyl (133 mg, 0.284 mmol) and cesium carbonate (694 mg, 2.13 mmol) were added under nitrogen protection and reacted at 100°C for 5 hours. The mixture was cooled to room temperature, diluted with water, and extracted with ethyl acetate. The organic phases were combined, and the mixture was dried over anhydrous sodium sulfate and subjected to column chromatography to obtain yellow solid compound 7-7.

Synthesis of compound 7-8:

Compound 7-7 (203 mg, 0.261 mmol) was dissolved in 5 ml of methanol, and 111 mg of 5% palladium carbon catalyst and ammonium formate (165 mg, 2.61 mmol) were added, and the mixture was heated to reflux overnight. The metal palladium was filtered off with diatomaceous earth, and the filtrate was dried by rotary evaporation, dissolved in methanol, and a formaldehyde aqueous solution (97 μL, 37% in H ₂ O, 1.31 mmol) and a drop of formic acid were added. Sodium cyanoborohydride (49 mg, 0.784 mmol) was slowly added at 0°C and reacted for one hour. Diluted with saturated sodium bicarbonate, extracted with ethyl acetate. The organic phases were combined, and the mixture was dried over anhydrous sodium sulfate, and then column chromatography was performed to obtain white solid compound 7-8.

Synthesis of compound S7:

Compound 7-8 (157 mg, 0.239 mmol) was dissolved in 2 ml of dichloromethane, and 2 ml of trifluoroacetic acid was slowly added under stirring. After the reaction solution was left to react at room temperature for 1 hour, the reaction solvent was removed by vacuum concentration. Then, 3 ml of dichloromethane was added to the mixture, the reaction system was placed at 0°C and N, N-diisopropylethylamine (119 μL, 0.717 mmol) was added, and acryloyl chloride (24 mg, 0.263 mmol) was slowly added dropwise under stirring. After the reaction solution was stirred at 0°C for 30 minutes, it was diluted with a saturated sodium bicarbonate aqueous solution and extracted with dichloromethane. The organic phases were combined, dried over anhydrous sodium sulfate, and after removing the solvent under reduced pressure, the mixture was separated by column chromatography to obtain a white solid compound S7. ¹ H NMR(400MHz,chloroform-d)δ7.76–7.59(m,2H),7.45–7.30(m,2H),7.29–7.18(m,2H),6.78–6.48(m,2H),6.40(dd,J＝16.7,1.8Hz,1H),5.82(d,J＝10.5Hz,1H ),5.42–4.55(m,1H),4.49–4.33(m,1H),4.13–3.80(m,3H),3.76–3.42(m,6H),3.38–2.96(m,5H),2.92–2.81(m,6H),2.50–2.26(m,4H).

Preparation Example 8 Preparation of Compound S8:

Compound S8 was synthesized by replacing the substrate with reference to compound Example S7. ¹ H NMR (400MHz, Methanol-d ₄ ) ^{1 H} NMR (400MHz, Methanol-d ₄ ) δ7.74–7.58(m,2H),7.49–7.17(m,4H),6.97–6.78(m,1H),6.67–6.55(m,1H),6.31(d,J＝16.7Hz,1H),5 .85(d,J＝10.6Hz,1H),5.39–5.13(m,1H),4.70–4.44(m,1H),4.37–4.24(m ,1H),4.21–3.56(m,10H),3.53–3.32(m,5H),3.30–2.85(m,6H),2.60(s,6 H).

Preparation Example 9 Preparation of Compound S9:

Synthesis of compound 9-2:

Compound 4-3 (4.2 g, 7.71 mmol) and compound 9-1 (3.73 g, 10.0 mmol) were dissolved in 35 ml of dry toluene, tri(dibenzylideneacetone)dipalladium (1.41 g, 1.54 mmol), 2-dicyclohexylphosphine-2',6'-diisopropoxy-1,1'-biphenyl (1.44 g, 3.08 mmol) and cesium carbonate (7.54 g, 23.1 mmol) were added under nitrogen protection and reacted at 100°C for 5 hours. The mixture was cooled to room temperature, diluted with water, and extracted with ethyl acetate. The organic phases were combined, and the mixture was dried over anhydrous sodium sulfate and subjected to column chromatography to obtain yellow solid compound 9-2.

Synthesis of compound S9:

Compound 9-2 (220 mg, 0.263 mmol) was dissolved in 2 ml of dichloromethane, and 2 ml of trifluoroacetic acid was slowly added under stirring. After the reaction solution was left to react at room temperature for 1 hour, the reaction solvent was removed by vacuum concentration. Then, 5 ml of dichloromethane was added to the mixture, the reaction system was placed at 0°C and N, N-diisopropylethylamine (131 μL, 0.790 mmol) was added, and acryloyl chloride (24 mg, 0.263 mmol) was slowly added dropwise under stirring. After the reaction solution was stirred at 0°C for 30 minutes, it was diluted with a saturated sodium bicarbonate aqueous solution and extracted with dichloromethane. The organic phases were combined, dried over anhydrous sodium sulfate, and after removing the solvent under reduced pressure, the mixture was separated by column chromatography to obtain a white solid compound S9. ¹ H NMR (400 MHz, DMSO-d ₆ )δ9.95–9.85(m,1H),7.91(d,J＝2.5Hz,1H),7.86–7.78(m,1H),7.64(dt,J＝6.3,3.2Hz,1H),7.33–7.21(m,2H),6.99–6.83(m,3H),6.52–6.44(m,1H) ,6.20(d,J＝16.9Hz,1H),5.78(d,J＝10.3Hz,1H),5.16–4.71(m,1H),4.53–4.00(m,2H),3.98–3.36(m,10H),3.21–2.80(m,5H),2.21–2.10(m,1H),1 .07–0.94(m,4H).

Preparation Example 10 Preparation of Compound S10:

The substrate was replaced and the compound S10 was synthesized according to the compound Example S9. ¹ H NMR (400 MHz, Methanol-d ₄ ) ¹ H NMR (400 MHz, Methanol-d ₄ )δ7.99–7.90(m,2H),7.57–7.49(m,1H),7.22–7.17(m,2H),6.94–6.76(m,3H),6.64–6.57(m,1H),6.29(d,J＝16.2Hz,1H),5.87–5.76(m,1H),5.23( s,1H),4.78–4.40(m,2H),4.20–3.72(m,3H),3.68–3.32(m,8H),3.23–3.00(m,2H),2.92–2.51(m,5H),2.15–2.05(m,1H),1.11–1.03(m,4H).

Preparation Example 11 Preparation of Compound S11:

The substrate was replaced and the compound S11 was synthesized by referring to the compound Example S9. ¹ H NMR (400 MHz, Methanol-d ₄ )δ8.03–7.94 (m, 2H),7.33–7.22 (m, 1H),7.18 (d, J=8.3 Hz, 1H),6.96–6.77 (m, 1H),6.72 (dd, J=12.9,7.4 Hz, 1H),6.63 (d, J=9.1 Hz, 1H),6.36–6.21 (m, 2H),5.90–5.77 (m, 1H),5 .27–5.13(m,1H),4.65–4.46(m,2H),3.89(d,J＝12.3Hz,1H),3.82–3.60(m,6H),3.59–3.31(m,5H),3.22–3.08(m,1H),3.00–2.82(m,2H),2.19–2. 04(m,1H),1.12–1.03(m,4H).

Preparation Example 12 Preparation of Compound S12:

Synthesis of compound S12:

Compound 4-4 (98 mg, 0.122 mmol) was dissolved in 1 mL of dichloromethane, and 1 mL of trifluoroacetic acid was slowly added under stirring. After the reaction solution was left to react at room temperature for 1 hour, the reaction solvent was removed by vacuum concentration. Then, 2 mL of N, N-dimethylformamide was added to the mixture, and N, N-diisopropylethylamine (61 μL, 0.367 mmol) was added, and O-(7-azabenzotriazole-1-yl)-N, N, N', N'-tetramethyluronium hexafluorophosphate (93 mg, 0.245 mmol) and 2-butynoic acid (13 mg, 0.159 mmol) were further added. After the reaction solution was stirred at room temperature for 2 hours, water and dichloromethane were added for extraction. The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the mixture was separated by column chromatography to obtain a yellow solid compound. ¹ H NMR (400MHz, Methanol-d ₄ ) δ8.02–7.91(m,3H),7.59–7.52(m,1H),7.34–7.29(m,1H),7.26–7.15(m,1H),6.98–6.62(m,3H),5.26–5.11(m,0.5H),4.79– 4.51(m,1H),4.40–4.17(m,2H),4.16–4.00(m,0.5H),3.94–3.32(m,10H),3.26–2.78(m,4H),2.18–2.01(m,4H),1.12–0.99(m,4H).

Preparation Example 13 Preparation of Compound S13:

The substrate was replaced and the compound S13 was synthesized by referring to the compound Example S12. ¹ H NMR (400 MHz, Methanol-d ₄ ) ¹ H NMR (400 MHz, Methanol-d ₄ )δ8.01–7.92(m,3H),7.65–7.55(m,1H),7.35–7.29(m,1H),7.26–7.17(m,1H),6.96–6.72(m,2H),6.69–6.57(m,1H),5.45–5.29(m,2H),5.27–5.1 2(m,0.5H),4.79–4.52(m,1H),4.38–4.18(m,2H),4.16–3.98(m,0.5H),3.94–3.36(m,10H),3.26–2.79(m,4H),2.17–2.04(m,1H),1.13–1.00(m,4 H).

Preparation Example 14 Preparation of Compound S14:

The substrate was replaced and the compound S14 was synthesized according to Example S4. ¹ H NMR (400 MHz, Methanol-d ₄ ) ¹ H NMR (400 MHz, Methanol-d ₄ ) δ8.68–8.57 (m, 1H), 8.07–7.92 (m, 2H), 7.62–7.53 (m, 1H), 7.26–7.15 (m, 1H), 6.98–6.72 (m, 3H), 6.62 (dd, J=8.7, 6.2 Hz, 1H), 6.29 (d, J=16.8 Hz, 1H), 5.88–5.74 (m ,1H),5.26–5.11(m,0.5H),4.79–4.51(m,1H),4.40–4.17(m,2H),4.16–4.00(m,0.5H),3.94–3.32(m,10H),3.26–2.78(m,4H),2.17–2.06(m,1H) ,1.14–1.02(m,4H).

Experimental Example:

1. Degradation activity of compounds on KRAS protein in pancreatic cancer cells MIA PaCa-2

MIA PaCa-2 is a pancreatic cancer cell with KRAS ^G12C mutation. This example will use this cell as an example to investigate the degradation activity of the compound on KRAS protein. MIA PaCa-2 was inoculated in a well plate and cultured overnight. The compound was administered at concentrations of 1nM, 10nM, 100nM, 1000nM, and 10000nM, respectively, and incubated at 37°C for 6h. After discarding the culture medium, the cells were lysed with RIPA lysis buffer, centrifuged at 10000×g for 10 minutes at 4°C, and the supernatant was taken for BCA protein concentration quantification. After adjusting the concentration with RIPA, 5× loading buffer was added and heated at 95°C for 10 minutes. Subsequently, the cell lysis sample was separated by SDS-PAGE gel and transferred to a nitrocellulose membrane. The membrane was blocked with 5% milk-TBST at room temperature for 1 hour, washed 3 times with TBST, and then incubated with primary antibodies such as Ras (#8955, CST), p-ERK ^T202/Y204 (#4370, CST), p-AKT (#4060, CST) and GAPDH (#2118, CST) at 4°C overnight, washed 3 times with TBST, and incubated with secondary antibodies (#7074, CST) for 1 hour, and then the bands were detected by chemiluminescence. The results were observed by a developer, and the Western results were quantified using software. GraphPad Prism 8.0 software was used to simulate and calculate the half-degradation concentration (DC ₅₀ ) of the compound for degradation of KRAS-12C mutant protein.

Table 1 KRAS protein degradation activity in tumor cells

Compound No. DC ₅₀ (μM) S1 B S2 B S3 A S4 A S5 B S6 A S7 B S8 C S9 A S10 A S11 A S12 B S13 B S14 A

Note: A refers to DC ₅₀ ≤1μM, B refers to 1μM<DC ₅₀ ≤5μM, C refers to DC ₅₀ >10μM

From the data in Table 1 above, it can be seen that most of the compounds have good K-RAS protein degradation activity, and their DC50 values are less than 5 μM; among them, the DC50 values of compounds S3, S4, S6, S9, S10, S11 and S14 are less than 1 μM.

Figure 1 shows the degradation effect of compound S9 on KRAS protein in MIA PaCa-2 and its effect on phosphorylation of downstream effector proteins.

As shown in FIG1 , compound S9 showed a dose-dependent degradation of KRAS protein, with a half-degradation concentration DC ₅₀ =60.49 nM; in addition, the compound was also able to inhibit the phosphorylation of downstream effector proteins of the KRAS pathway.

2. Proliferation inhibitory activity of compounds on pancreatic cancer cell MIA PaCa-2

MIA PaCa-2 is a pancreatic cancer cell with KRAS ^G12C mutation. This example will take this cell as an example to investigate the inhibitory activity of the compound on KRAS mutant tumor cells. Take the MIAPaCa-2 cells in the logarithmic growth phase and inoculate them in a 12-well plate. Inoculate 1 mL of a suspension containing 2,000 cells in each well and culture them overnight in a 37°C, 5% CO ₂ incubator. After the cells are completely attached to the wall, add different drugs for treatment. After 3 days of drug treatment, discard the culture medium, add 1 mL of 4% paraformaldehyde to each well and fix it in the dark for 20 minutes, then add 500 μL of 0.2% crystal violet to each well for staining for 10 minutes, wash the staining solution with tap water, and leave it at room temperature to dry. After drying, use 10% acetic acid to dissolve the crystal violet, transfer it to 96 wells, and detect the absorbance of each well at a wavelength of 595 nm using an ELISA reader. The cell survival rate of the control group (i.e., the group without drug treatment) was taken as 100%, and the absorbance values of the other groups were compared with the absorbance value of the control group to calculate the corresponding survival rates, and the IC ₅₀ values were simulated and calculated using GraphPad Prism 8.0 software.

Table 2 Tumor cell proliferation inhibitory activity

Compound IC ₅₀ (μM) S1 B S2 B S3 A S4 A S5 B S6 A S7 B S8 C S9 A S10 A S11 A S12 B S13 B S14 A

Note: A refers to IC ₅₀ ≤1μM, B refers to 1μM<IC ₅₀ ≤5μM, C refers to IC ₅₀ >10μM

As can be seen from Table 2, most of the compounds showed good tumor cell proliferation inhibitory activity, and their _IC50 values were less than 5 μM; among them, the _IC50 values of S3, S4, S6, S9, S10, S11 and S14 were less than 1 μM.

FIG2 is a graph showing the growth inhibition rate of compound S9 on MIA PaCa-2 cells, with IC ₅₀ =0.34±0.04 μM.

3. In vivo pharmacodynamics experiments

In this example, the mouse transplanted tumor of human pancreatic cancer MIA PaCa-2 cells was used as a model to evaluate the in vivo anti-KRAS mutant tumor activity of the compound.

1. Experimental methods:

Cell culture: Human pancreatic cancer MIA PaCa-2 cells were cultured in monolayer in vitro. The culture conditions were 10% fetal bovine serum and 2.5% horse serum in DMEM medium and cultured in a 37°C 5% carbon dioxide cell culture incubator. Every 2-3 days, trypsin (containing EDTA) was used for routine digestion and passage. When the cell density in the culture medium was 70%-80% and the number reached the experimental requirements, the cells were collected, counted, and resuspended in an appropriate amount of DMEM medium to prepare a cell suspension with a cell density of 4×107 cells/mL.

Cell inoculation: 0.125 mL, i.e., 5×106 MIA PaCa-2 cells per mouse, was subcutaneously inoculated into the lower right armpit of each mouse.

Experimental operation: ^When the average tumor volume reached 100 mm3, the mice were randomly divided into groups according to the tumor volume, with 6 mice in each group. The blank group was given a dose of 0, and the test groups were given doses of 30 mg/kg and 60 mg/kg, respectively. The dosing volume was 0.2 mL, and the drugs were administered by intraperitoneal injection. The experimental period was 21 days. The drugs were administered twice a week in the first two weeks, for a total of four times, and then the drugs were stopped and observed for another week.

2. Tumor volume, mouse body weight measurement and experimental indicators:

Tumor volume and mouse body weight were measured every two days.

The tumor diameter was measured with a vernier caliper, and the mouse tumor volume was calculated using the formula: V = a × b2 × 0.5, where a and b represent the long and short diameters of the tumor, respectively.

The anti-tumor efficacy of the compound is evaluated by TGI (%) or relative tumor proliferation rate T/C (%). Relative tumor proliferation rate T/C (%) = TRTV/CRTV × 100% (TRTV: RTV of the treatment group; CRTV: RTV of the negative control group). The relative tumor volume (RTV) is calculated based on the results of the swelling and pain measurement. The calculation formula is RTV = Vt/V0, where V0 is the average tumor volume measured at the time of group administration (i.e. D0), Vt is the average tumor volume at a certain measurement, and TRTV and CRTV are taken from the same day. TGI (%) reflects the tumor growth inhibition rate. TGI (%) = [(1-(average tumor volume at the end of administration of a treatment group - average tumor volume at the beginning of administration of the treatment group))/(average tumor volume at the end of treatment of the solvent control group - average tumor volume at the beginning of treatment of the solvent control group)] × 100%.

Statistical method: Two-way ANOVA, ** represents P < 0.01, *** represents P < 0.001

3. Experimental results:

FIG3 is a curve diagram showing the in vivo tumor volume inhibition effect of compound S9 in a nude mouse xenograft tumor model of pancreatic cancer MIA PaCa-2 cells.

As shown in Figure 3, compound S9 exhibited good in vivo efficacy in the MIA PaCa-2 cell nude mouse transplant tumor efficacy model, wherein the tumor growth inhibition rate (TGI) was 73.2% at a dose of 60 mg/kg injected intraperitoneally, twice a week, and a total of four times.

All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference individually. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims (10)

1. A compound is characterized in that the compound is a compound shown in a formula I or a pharmaceutically acceptable salt or solvate thereof,

Wherein each R ¹、R² is independently selected from the group consisting of: hydrogen, halogen, hydroxy, amino, cyano, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 3-8 membered heterocycloalkyl containing 1,2 or 3 heteroatoms selected from N, O or S, substituted or unsubstituted C5-C10 aryl, substituted or unsubstituted 5-10 membered heteroaryl containing 1,2 or 3 heteroatoms selected from N, O or S; each of the substitutions independently refers to substitution with 1,2,3, 4, 5 or 6 substituents selected from the group consisting of: - (c=o) -NRaRb, C1-C6 alkyl, halo C1-C6 alkyl, C1-C6 alkoxy substituted C1-C6 alkyl, C1-C6 alkoxy, oxo (=o), halogen, C3-C8 cycloalkyl, 3-8 membered heterocycloalkyl containing 1,2 or 3 heteroatoms selected from N, O or S, C5-C10 aryl, 5-10 membered heteroaryl containing 1,2 or 3 heteroatoms selected from N, O or S;

R ³ is selected from the group consisting of: substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl; each of the substitutions independently refers to substitution with 1,2, 3, 4,5 or 6 substituents selected from the group consisting of: halogen, hydroxy, -NRaRb, 4-6 membered heterocycloalkyl containing 1,2 or 3 heteroatoms selected from N, O or S;

R ⁴ is selected from the group consisting of: a substituted or unsubstituted C5-C10 aryl, a substituted or unsubstituted 5-10 membered heteroaryl containing 1,2 or 3 heteroatoms selected from N, O or S; each of the substitutions independently refers to substitution with 1,2,3, 4, 5 or 6 substituents selected from the group consisting of: halogen, hydroxy, amino, C1-C6 alkyl, halogenated C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C3-C8 cycloalkyl;

Ra, rb are each independently selected from the group consisting of: hydrogen, C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl, 3-8 membered heterocycloalkyl containing 1,2 or 3 heteroatoms selected from N, O or S, C5-C10 aryl, 5-10 membered heteroaryl containing 1,2 or 3 heteroatoms selected from N, O or S.

2. The compound of claim 1, wherein each R ¹、R² is independently selected from the group consisting of: hydrogen, halogen, hydroxy, amino, cyano, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 3-8 membered heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, substituted or unsubstituted C5-C10 aryl, substituted or unsubstituted 5-10 membered heteroaryl containing 1, 2 or 3 heteroatoms selected from N, O or S; each of the substitutions independently refers to substitution with 1, 2,3,4, 5 or 6 substituents selected from the group consisting of: - (c=o) -NRaRb, C1-C6 alkyl, halo C1-C6 alkyl, C1-C6 alkoxy substituted C1-C6 alkyl, C1-C6 alkoxy, oxo (=o), halogen;

ra, rb are as defined in claim 1.

3. The compound of claim 1, wherein R ³ is selected from the group consisting of: substituted or unsubstituted C2-C4 alkenyl, substituted or unsubstituted C2-C4 alkynyl; each of the substitutions independently refers to substitution with 1,2, 3, 4, 5 or 6 substituents selected from the group consisting of: halogen, hydroxy, -NRaRb;

ra, rb are as defined in claim 1.

4. The compound of claim 1, wherein R ⁴ is selected from the group consisting of: a substituted or unsubstituted C8-C10 aryl, a substituted or unsubstituted 8-10 membered heteroaryl containing 1,2 or 3 heteroatoms selected from N, O or S; each of the substitutions independently refers to substitution with 1,2, 3, 4, 5 or 6 substituents selected from the group consisting of: halogen, hydroxy, amino, C1-C6 alkyl, halogenated C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C3-C8 cycloalkyl.

5. The compound of claim 1, wherein Ra, rb are each independently selected from the group consisting of: hydrogen, C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl.

6. The compound of claim 1, wherein the compound is selected from the group consisting of:

7. a pharmaceutical composition comprising a pharmaceutically acceptable carrier and one or more safe and effective amounts of a compound of claim 1.

8. Use of a compound according to claim 1 for the preparation of a medicament for the prevention and/or treatment of KRAS mutation related diseases.

9. The use of claim 8, wherein the KRAS mutation is selected from the group consisting of: KRAS ^G12C mutation, KRAS ^G12D mutation.

10. The use of claim 8, wherein the KRAS mutation related disease is selected from the group consisting of: pancreatic cancer, colorectal cancer, lung adenocarcinoma, lung cancer, cholangiocarcinoma, cervical cancer, bladder cancer, liver cancer, breast cancer, endometrial cancer, skin cancer, ovarian cancer, gastric cancer, urinary tract cancer, soft tissue sarcoma, esophageal cancer, multiple myeloma, prostate cancer, renal cancer, gastrointestinal neuroendocrine tumor, gastrointestinal stromal tumor, head and neck cancer, glioma, salivary gland cancer, bone cancer, anal cancer, thyroid cancer, melanoma, mature B-cell tumor, small intestine cancer, ampulla cancer.