Treatment of conditions using mutant P53 reactivation compounds

Abstract

Mutations in oncogenes and tumor suppressor factors contribute to the development and progression of cancer. This disclosure describes compounds and methods for restoring DNA-binding affinity of p53 mutants, as well as their use in diagnostic assays to guide the treatment of subjects with said compounds for cancer. The compounds of this disclosure can bind to mutant p53 and restore the ability of p53 mutants to bind to DNA and activate downstream effectors involved in tumor suppression. The disclosed compounds can be used to reduce the progression of cancers containing p53 mutations.

Description

[0001] Cross-referencing This application claims the benefits of U.S. Provisional Patent Application No. 63 / 669,209, filed July 9, 2024; U.S. Provisional Patent Application No. 63 / 632,458, filed April 10, 2024; U.S. Provisional Patent Application No. 63 / 566,417, filed March 18, 2024; and U.S. Provisional Patent Application No. 63 / 589,229, filed October 10, 2023, each of which is incorporated herein by reference in its entirety.

[0002] background Cancer, an uncontrolled proliferation of cells, is a multifactorial disease characterized by tumor formation, growth, and, in some cases, metastasis. Cells carrying activated oncogenes, damaged genomes, or other cancer-promoting alterations can be prevented from replicating through a sophisticated tumor suppressor network. The most important component of this tumor suppressor network is p53, one of the most potent tumor suppressor factors in cells. Both the wild-type and mutant conformations of p53 are associated with cancer progression.

[0003] By incorporating references All publications, patents and patent applications mentioned in this specification are incorporated herein by reference as if each individual publication, patent and patent application were specifically and individually indicated to be incorporated by reference.

[0004] Overview In some embodiments, this disclosure provides a method for treating a condition in a subject with a corresponding need, the method comprising administering to the subject a therapeutically effective amount of a small molecule compound that binds to a p53 mutant protein, wherein the mutation status of the KRAS gene in the subject has been previously determined.

[0005] In some embodiments, this disclosure provides a method for treating a subject with a corresponding need for treatment, the method comprising: a) determining the mutation status of the KRAS gene in the subject based on the results of a measurement performed on a biological sample of the subject; and b) administering to the subject a therapeutically effective amount of a compound binding to the p53 mutant protein, based at least in part on the mutation status of the KRAS gene in the subject. Brief description of the attached diagram Figure 1 The figure illustrates the dose escalation protocol used in studies evaluating compound 1 in cancer treatment.

[0007] Figure 2 This is a graph illustrating the exposure of compound 1 to human subjects on day 15 (steady state).

[0008] Figure 3 It is a summary TP53 Y220C / KRAS Wild-type potency can be evaluated by plotting the reduction in target lesions across tumor types in the population (in patients receiving 1150 mg QD to 1500 mg BID compound 1).

[0009] Figure 4 The diagram illustrates the effects of compound 1 therapy. TP53 Y220C / KRAS Wild-type potency can be evaluated by plotting the duration of efficacy in patients within a population.

[0010] Figure 5 This is a graph summarizing the percentage change in the TP53 Y220C variant allele frequency relative to baseline at week 3 of treatment with compound 1.

[0011] Figure 6 Images of patients with triple-negative breast cancer at baseline and after cycle 3 of compound 1 treatment are provided.

[0012] Figure 7 This is a graph summarizing the best percentage change in the total number of tumor target lesions among 15 patients in the ovarian subgroup with measurable disease.

[0013] Figure 8 This is a graph summarizing the best percentage change in CA-125 levels in 15 patients in a subgroup of ovarian cysts with measurable CA-125 at baseline.

[0014] Figure 9 This is a graph summarizing response time and treatment duration for patients with a post-baseline tumor assessment ≥1 in the ovarian cyst subgroup with measurable disease at baseline.

[0015] Figure 10 Linear graphs of plasma concentrations of 1000 mg of compound 1 over time are shown, under conditions of feeding or fasting.

[0016] Figure 11 A semi-logarithmic plot of plasma concentrations of 1000 mg of compound 1 over time is shown, under conditions of feeding or fasting.

[0017] Figure 12 Linear graphs of plasma concentrations of 2000 mg of compound 1 over time are shown, depending on race, under fed or fasted conditions.

[0018] Figure 13 A semi-logarithmic plot of plasma concentrations of 2000 mg of compound 1 over time is shown, depending on race, under fed or fasted conditions.

[0019] Figure 14The plasma pharmacokinetic parameters (AUC) of 1000 mg of compound 1 are shown in both fed and fasted states. 0-inf The trapezoidal diagram.

[0020] Figure 15 The plasma pharmacokinetic parameters (AUC) of 1000 mg of compound 1 are shown in both fed and fasted states. 0-last The trapezoidal diagram.

[0021] Figure 16 The plasma pharmacokinetic parameters (AUC) of 1000 mg of compound 1 are shown in both fed and fasted states. 0-last The trapezoidal diagram.

[0022] Figure 17 The AUC values ​​of 2000 mg compound 1 in plasma were shown in Caucasian / non-Asian subjects under both eating and fasting conditions. 0-inf The trapezoidal diagram.

[0023] Figure 18 The AUC values ​​of 2000 mg compound 1 in plasma were shown in Caucasian / non-Asian subjects under both eating and fasting conditions. 0-last The trapezoidal diagram.

[0024] Figure 19 The plasma pharmacokinetic parameters C of compound 1 at 2000 mg were shown in Caucasian / non-Asian subjects under both eating and fasting conditions. max The trapezoidal diagram.

[0025] Figure 20 A graph showing the arithmetic mean cumulative urinary excretion fraction (%) of 1000 mg of compound 1 over time under eating or fasting conditions is presented.

[0026] Figure 21 The graph shows the arithmetic mean cumulative urinary excretion fraction (%) of 1000 mg compound 1 over time in Caucasian / non-Asian subjects under eating or fasting conditions.

[0027] Figure 22 The graph shows the arithmetic mean cumulative urinary excretion fraction (%) of Japanese subjects who consumed 1000 mg of compound 1 over time while fasting.

[0028] Detailed description This disclosure provides, for example, methods for restoring wild-type function of mutant p53 using compounds disclosed herein. The compounds can bind to mutant p53 and restore the ability of the p53 mutant to bind to DNA. Restoration of the activity of the p53 mutant can allow activation of downstream effectors of p53, leading to inhibition of cancer progression. This disclosure also provides methods for treating cancerous lesions or tumors carrying p53 mutations.

[0029] Cancer is a group of related diseases characterized by the uncontrolled proliferation of cells with the potential to metastasize throughout the body. Cancers can be classified into five main categories, including, for example: carcinomas, which can arise from cells covering both internal and external parts of the body, such as the lungs, breast, and colon; sarcomas, which can arise from cells located in bone, cartilage, fat, connective tissue, muscle, and other supporting tissues; lymphomas, which can arise in lymph nodes and immune system tissues; leukemia, which can arise in the bone marrow and accumulate in the bloodstream; and adenomas, which can arise in the thyroid gland, pituitary gland, adrenal gland, and other glandular tissues.

[0030] Although different cancers can develop in almost any body tissue and have unique characteristics, the basic processes that lead to cancer are likely similar in all forms of disease. Cancer begins when cells break free from the normal constraints on cell division and begin to grow and divide uncontrollably. Gene mutations in cells can prevent cells from repairing damaged DNA or initiating apoptosis, and can lead to uncontrolled cell growth and division.

[0031] The ability of tumor cell populations to proliferate is determined not only by the rate of cell proliferation but also by the rate of cell attrition. Programmed cell death, or apoptosis, represents the main mechanism of cell attrition. Cancer cells can evade apoptosis through various strategies, such as inhibiting p53 function, thereby suppressing the expression of pro-apoptotic proteins.

[0032] Oncogenes and tumor suppressor genes regulate cell proliferation. Gene mutations can affect oncogenes and tumor suppressor factors, potentially abnormally activating or inhibiting their activity, further promoting uncontrolled cell division. While oncogenes contribute to cell growth, tumor suppressor genes slow cell division by repairing damaged DNA and activating apoptosis. Oncogenes that can mutate in cancer include, for example, Cdk1, Cdk2, Cdk3, Cdk4, Cdk6, EGFR, PDGFR, VEGF, HER2, Raf, KRAS, and myc. Tumor suppressor genes that can mutate in cancer include, for example, BRCA1, BRCA2, cyclin-dependent kinase inhibitor 1C, retinoblastoma protein (pRb), PTEN, p16, p27, p53, and p73.

[0033] K-Ras K-Ras are Ras family GTPases encoded by the KRAS oncogene. In mammals, KRAS can be expressed via selective splicing as either of two isotypes, K-Ras4A and K-RasB. In wild-type K-Ras, GTP binding activates intracellular signaling pathways that play a role in cell proliferation and angiogenesis. Oncogenic mutations in the KRAS gene can constitutively activate K-Ras, thereby conferring ligand-independent activation that leads to tumorigenesis. Cancer-associated KRAS mutations include those listed in Table 1.

[0034] Table 1 In some embodiments, the p53-binding compounds of this disclosure are more effective in treating cancers in subjects without oncogenic KRAS mutations than in treating cancers in subjects with cancers containing oncogenic KRAS mutations. In some embodiments, subjects are selected for treatment with the p53-activating compounds of this disclosure based at least in part on the absence of mutations in codons 12, 13, and 61 of the KRAS gene in the subject. In some embodiments, subjects are selected for treatment based on the absence of mutations in codons 12, 13, 59, 61, 117, and 146 of the KRAS gene in the subject. In some embodiments, the KRAS gene in the subject is the gene of cancer cells in the subject.

[0035] In some implementations, the subject's cancer cells contain the KRAS gene expressing the wild-type K-ras protein. The wild-type K-ras protein can be, for example, K-Ras4A. , Or K-Ras4B: .

[0036] BRCA1 / 2 BRCA1 and BRCA2 are respectively composed of BRCA1 and BRCA2The tumor suppressor gene encodes a human protein. Both proteins are involved in repairing double-strand breaks in DNA. In some embodiments, the p53-binding compounds of this disclosure are more effective in treating cancers in subjects without oncogenic BRCA1 or BRCA2 mutations than in treating cancers in subjects with oncogenic BRCA1 or BRCA2 mutations. In some embodiments, the p53-binding compounds of this disclosure are more effective in treating cancers in subjects without oncogenic BRCA1 mutations than in treating cancers in subjects with oncogenic BRCA2 mutations.

[0037] PIK3CA The PIK3CA gene encodes the p110 alpha (p110α) protein, which forms the catalytic subunit of phosphatidylinositol 3-kinase (PI3K). Mutations in the p110α subunit can be found in many forms of cancer, including ovarian cancer, head and neck cancer, lung cancer, skin cancer, brain cancer, breast cancer, prostate cancer, colorectal cancer, cervical cancer, esophageal cancer, skin cancer, thyroid cancer, gastric cancer, liver cancer, and pancreatic cancer, and often co-occur with HER-2 protein overexpression in breast cancer. In some embodiments, the p53-binding compounds of this disclosure are more effective in treating cancers in subjects without oncogenic PIK3CA mutations than in treating cancers in subjects with cancer-causing PIK3CA mutations. Cancer-associated PIK3CA mutations include those listed in Table 2.

[0038] Table 2 tumor suppressor p53 The tumor suppressor protein p53 is a 393-amino acid transcription factor that regulates cell growth in response to cellular stresses, including, for example, UV radiation, hypoxia, oncogene activation, and DNA damage. p53 possesses various mechanisms for inhibiting cancer progression, including, for example, initiating apoptosis, maintaining genomic stability, cell cycle arrest, inducing senescence, and inhibiting angiogenesis. Due to its crucial role in tumor suppression, p53 is inactivated in almost all cancers either through direct mutations or by perturbations of related signaling pathways involved in tumor suppression. Homozygous loss of the p53 gene occurs in almost all types of cancer, including breast, colon, and lung cancer. The presence of certain p53 mutations in several types of human cancers may be associated with a less favorable patient prognosis.

[0039] In the absence of stress signals, p53 levels are maintained at low levels via the interaction between p53 and Mdm2 (E3 ubiquitin ligase). In non-stressed cells, Mdm2 can target p53 for degradation by the proteasome. Under stress conditions, the interaction between Mdm2 and p53 is disrupted, and p53 accumulates. A key event leading to p53 activation is the phosphorylation of the N-terminal domain of p53 by protein kinases, thereby transducing upstream stress signals. Phosphorylation of p53 leads to a conformational change, which can promote DNA binding of p53 and allow transcription of downstream effectors. Activation of p53 can induce, for example, intrinsic apoptosis pathways, extrinsic apoptosis pathways, cell cycle arrest, senescence, and DNA repair. p53 can activate proteins involved in the aforementioned pathways, including, for example, Fas / Apo1, Killer / DR5, Bax, Puma, Noxa, Bid, caspase-3, caspase-6, caspase-7, caspase-8, caspase-9, and p21(WAF1). Additionally, p53 can indirectly inhibit the transcription of various genes, including, for example, c-MYC, cyclin B, VEGF, RAD51, and hTERT.

[0040] Each strand of the p53 tetramer comprises several functional domains, including a transactivation domain (amino acids 1-100), a DNA-binding domain (amino acids 101-306), and a tetramerization domain (amino acids 307-355), which are highly mobile and largely unstructured. Most p53 cancer mutations reside in the protein's DNA-binding core domain, which contains a central β-sandwich structure of antiparallel β-sheets, serving as the basic scaffold of the DNA-binding surface. The DNA-binding surface contains two β-turn loops, L2 and L3 (stabilized by zinc ions, e.g., at Arg175 and Arg248) and a loop-sheet-helical motif. In summary, these structural elements form an extended DNA-binding surface enriched with positively charged amino acids and specifically contacting various p53 responsive elements.

[0041] Because p53 mutations are prevalent in almost every type of cancer, reactivation of wild-type p53 function in cancer cells could be an effective therapy. Mutations in p53, located at the DNA-binding domain or periphery of the DNA-binding surface of a protein, can lead to abnormal protein folding or reduced DNA-binding affinity required for DNA recognition and binding. p53 mutations can occur at, for example, amino acids Val143, His168, Arg175, Tyr220, Gly245, Arg248, Arg249, Phe270, Arg273, and Arg282. p53 mutations that can eliminate p53 activity include, for example, R175H, Y220C, G245S, R248Q, R248W, R273C, R273H, and R282H. p53 mutations can distort the structure of DNA-binding sites, causing folded proteins to become thermodynamically destabilized at body temperature, or weakening consensus DNA binding. Wild-type function of p53 mutants can be restored by binding p53 mutants to compounds that shift the fold-unfold equilibrium toward the folded state, thereby reducing the rate of unfolding and destabilization; or by conjugating small molecules to the DNA binding interface to restore common DNA binding.

[0042] Non-limiting examples of amino acids include: alanine (A, Ala); arginine (R, Arg); asparagine (N, Asn); aspartic acid (D, Asp); cysteine ​​(C, Cys); glutamic acid (E, Glu); glutamine (Q, Gln); glycine (G, Gly); histidine (H, His); isoleucine (I, Ile); leucine (L, Leu); lysine (K, Lys); methionine (M, Met); phenylalanine (F, Phe); proline (P, Pro); serine (S, Ser); threonine (T, Thr); tryptophan (W, Trp); tyrosine (Y, Tyr); and valine (V, Val).

[0043] CA-125 CA-125 is a cancer antigen. A CA-125 test can be used to measure the amount of CA-125 protein in a patient's blood. A CA-125 test can be used to monitor certain cancers during and after treatment. In some embodiments, this disclosure provides a method of treating cancer, comprising: a) administering a therapeutically effective amount of a compound of this invention (e.g., compound 1) to a subject in appropriate need; and b) monitoring the level of CA-125 protein in the subject after administration. In some embodiments, this disclosure provides a method including monitoring the level of CA-125 in a subject undergoing cancer therapy with a compound of this invention (e.g., compound 1).

[0044] HRD Homologous recombination is the process in which nucleotide sequences are exchanged between different strands of DNA. Homologous recombination involves many different biological processes, such as occurring as part of DNA repair processes (e.g., double-strand break repair pathways and synthesis-dependent strand annealing pathways) and during meiosis / gametosis in eukaryotic organisms. Homologous recombination defects, also known as homologous recombination repair defects, homologous repair deficiencies, or HRD, are reductions or impairments in the homologous recombination process. Samples suffering from HRD can accumulate genomic errors or chromosomal aberrations, which can be used as biomarkers of HRD. Chromosomal aberrations (CA) are detectable variations in the chromosomal DNA of a sample. In some embodiments, CA falls into at least one of three overlapping categories: loss of heterozygosity (LOH), allele imbalance (e.g., telomere allele imbalance (TAI)), or large-scale shift (LST).

[0045] In some embodiments, HRD status is determined by detecting chromosomal aberrations (CA) in a sample obtained from a patient (e.g., a tumor sample). In some embodiments, a sample with a positive HRD status satisfies or exceeds a threshold number or level of CA at a specified number of chromosomal indicator regions. In some embodiments, a commercially available diagnostic is used to determine HRD status to detect chromosomal aberrations in a sample (e.g., a tumor sample) and / or to assess whether the sample is capable of repairing double-strand DNA breaks. Non-limiting examples of diagnostic agents suitable for assessing HRD status include myChoiceHRD. TM Diagnostic kit (Myriad Genetics, South San Francisco).

[0046] In some implementations, the p53-binding compounds of this disclosure are more effective in treating cancers in HRD-negative subjects than in treating cancers in HRD-positive subjects.

[0047] The mechanism of the compounds disclosed herein The compounds of this disclosure can selectively bind to p53 mutants and can restore wild-type activity of p53 mutants, including, for example, DNA binding function and activation of downstream targets involved in tumor suppression. In some embodiments, the compounds of this disclosure selectively bind to the p53 Y220C mutant. The Y220C mutant is a temperature-sensitive mutant that binds to DNA at lower temperatures and denatures at body temperature. The compounds of this disclosure can stabilize the Y220C mutant to reduce the likelihood of protein denaturation at body temperature.

[0048] In some embodiments, the compounds of this disclosure stabilize mutant p53 and allow mutant p53 to bind to DNA, thereby shifting the balance of wild-type and mutant p53 proteins to wild-type p53. In some embodiments, the compounds of this disclosure reactivate mutant p53 protein to provide wild-type p53 activity. In some embodiments, the compounds of this disclosure reactivate mutant p53 protein to provide pro-apoptotic p53 activity. In some embodiments, the compounds of this disclosure reactivate mutant p53 protein to block angiogenesis. In some embodiments, the compounds of this disclosure reactivate mutant p53 protein to induce cellular senescence. In some embodiments, the compounds of this disclosure reactivate mutant p53 protein to induce cell cycle arrest.

[0049] Located on the periphery of the p53 β-sandwich structure connecting β-chains S7 and S8, the aromatic ring of Y220 is an integral part of the hydrophobic core of the β-sandwich structure. Due to the formation of internal surface cavities, the Y220C mutation can be highly unstable. The compounds of this disclosure can bind to and occupy these surface cavities to stabilize the β-sandwich structure, thereby restoring wild-type p53 DNA binding activity.

[0050] To determine the ability of the compounds of this disclosure to bind to and stabilize mutant p53, assays can be employed to detect, for example, conformational changes in the p53 mutant or activation of the wild-type p53 target. Conformational changes in p53 can be measured by, for example, differential scanning fluorometry (DSF), isothermal titration calorimetry (ITC), nuclear magnetic resonance spectroscopy (NMR), or X-ray crystallography. Additionally, antibodies specific to the wild-type or mutant conformations of p53 can be used to detect conformational changes via, for example, immunoprecipitation (IP), immunofluorescence (IF), or Western blotting.

[0051] Methods for detecting the ability of p53 mutants to bind DNA may include, for example, DNA affinity immunoblotting, modified enzyme-linked immunosorbent assay (ELISA), electrophoretic mobility shift assay (EMSA), fluorescence resonance energy transfer (FRET), homogeneous time-resolved fluorescence (HTRF), and chromatin immunoprecipitation (ChIP) assays.

[0052] To determine whether the compounds described herein can reactivate the transcriptional activity of p53, activation of downstream targets in the p53 signaling cascade can be measured. Activation of p53 effector proteins can be detected by, for example, immunohistochemistry (IHC-P), reverse transcription polymerase chain reaction (RT-PCR), and Western blotting. p53 activation can also be measured by inducing apoptosis via a caspase cascade and using methods including, for example, annexin V staining, TUNEL assay, pro-caspase and caspase levels, and cytochrome c levels. Another consequence of p53 activation is senescence, which can be measured using methods such as β-galactosidase staining.

[0053] The p53 mutant that can be used to determine the effectiveness of the compounds of this disclosure in enhancing the DNA-binding ability of p53 mutants is a p53 truncated mutant containing only amino acids 94-312, including the DNA-binding domain of p53. For example, the sequence of the p53 Y220C mutant used to test the efficacy of the compounds could be: Compared to the ability of p53 mutants to bind DNA in the absence of compounds of this disclosure, compounds of this disclosure can enhance the ability of p53 mutants to bind DNA by at least or up to about 0.1%, at least or up to about 0.2%, at least or up to about 0.3%, at least or up to about 0.4%, at least or up to about 0.5%, at least or up to about 0.6%, at least or up to about 0.7%, at least or up to about 0.8%, and at least or up to about 0.9%, at least or up to about 1%, at least or up to about 2%, at least or up to about 3%, at least or up to about 4%, at least or up to about 5%, at least or up to about 6%, at least or up to about 7%, at least or up to about 8%, at least or up to about 9%, at least or up to about 10%, at least or up to about 11%, at least or up to about 12%, at least or up to about 13%, at least or up to about 14%, at least or up to about 15%, at least or up to about 16%, at least or up to about 17%, at least or up to about 18%, at least or up to about 19%, at least or up to about 20%, at least or up to about 21%, at least or up to about 22%, at least or up to about 23%, at least or up to about 24%, at least or up to about 25%, at least or up to about 26%, at least or up to about 27%, at least or up to about 28%, at least or up to about 29%, at least or up to about 30%, at least or up to about 31%, at least or up to about 32%, at least or up to about 33%, at least or up to about 34%, at least or up to about 35%, at least or up to about 36%, at least or up to about 37%, at least or up to about 38%, at least or up to about 39%, at least or up to about 40%, at least or up to about 41%, at least or up to about 42%, at least or up to about 43%, at least or up to about 44%, at least or up to about 45%, at least or up to about 46%, at least or up to about 47%, at least or up to about 48%, at least or up to about 4 9%, at least or up to about 50%, at least or up to about 51%, at least or up to about 52%, at least or up to about 53%, at least or up to about 54%, at least or up to about 55%, at least or up to about 56%, at least or up to about 57%, at least or up to about 58%, at least or up to about 59%, at least or up to about 60%, at least or up to about 61%, at least or up to about 62%, at least or up to about 63%, at least or up to about 64%, at least or up to about 65%, at least or up to about 66%, at least or up to about 67%, at least or up to about 68%, at least or up to about 69%, at least or up to about 70%, at least or up to about 71%, at least or up to about 72%, at least or up to about 7 3%, at least or up to about 74%, at least or up to about 75%, at least or up to about 76%, at least or up to about 77%, at least or up to about 78%, at least or up to about 79%, at least or up to about 80%, at least or up to about 91%, at least or up to about 92%, at least or up to about 93%, at least or up to about 94%, at least or up to about 95%, at least or up to about 96%, at least or up to about 97%, at least or up to about 98%, at least or up to about 99%, at least or up to about 100%, at least or up to about 125%, at least or up to about 150%, at least or up to about 175%, at least or up to about 200%, at least or up to about 225%, or at least or up to about 250%.

[0054] Compared to the activity of p53 mutants in the absence of the compound, the compounds described herein can increase the activity of p53 mutants by, for example, at least or up to about 2-fold, at least or up to about 3-fold, at least or up to about 4-fold, at least or up to about 5-fold, at least or up to about 6-fold, at least or up to about 7-fold, at least or up to about 8-fold, at least or up to about 9-fold, at least or up to about 10-fold, at least or up to about 11-fold, at least or up to about 12-fold, at least or up to about 13-fold, at least or up to about 14-fold, at least or up to about 15-fold, at least or up to about 16-fold, at least or up to about 17-fold. At least or up to about 18 times, at least or up to about 19 times, at least or up to about 20 times, at least or up to about 25 times, at least or up to about 30 times, at least or up to about 35 times, at least or up to about 40 times, at least or up to about 45 times, at least or up to about 50 times, at least or up to about 65 times, at least or up to about 70 times, at least or up to about 75 times, at least or up to about 80 times, at least or up to about 85 times, at least or up to about 90 times, at least or up to about 95 times, at least or up to about 100 times, at least or up to about 110 times, at least or up to about 120 times, at least or Up to approximately 130 times, at least or up to approximately 140 times, at least or up to approximately 150 times, at least or up to approximately 160 times, at least or up to approximately 170 times, at least or up to approximately 180 times, at least or up to approximately 190 times, at least or up to approximately 200 times, at least or up to approximately 250 times, at least or up to approximately 300 times, at least or up to approximately 350 times, at least or up to approximately 400 times, at least or up to approximately 450 times, at least or up to approximately 500 times, at least or up to approximately 550 times, at least or up to approximately 600 times, at least or up to approximately 650 times, at least or up to approximately 700 times, at least Or up to about 750 times, at least or up to about 800 times, at least or up to about 850 times, at least or up to about 900 times, at least or up to about 950 times, at least or up to about 1,000 times, at least or up to about 1,500 times, at least or up to about 2,000 times, at least or up to about 3,000 times, at least or up to about 4,000 times, at least or up to about 5,000 times, at least or up to about 6,000 times, at least or up to about 7,000 times, at least or up to about 8,000 times, at least or up to about 9,000 times, or at least or up to about 10,000 times.

[0055] The compounds of this disclosure can be used, for example, to induce apoptosis, cell cycle arrest, or cellular senescence. In some embodiments, the cells are cancer cells. In some embodiments, the cells carry a p53 mutation. Patients can be treated with the compounds of this disclosure based on whether they have platinum-sensitive, platinum-resistant, or platinum-refractory cancer. Platinum-resistant cancer includes, for example, cancer that progresses within 6 months of treatment with platinum-based chemotherapy (such as with cisplatin or carboplatin). Platinum-refractory cancer includes, for example, cancer that progresses during treatment with platinum-based chemotherapy or within four weeks of the last dose of platinum-based chemotherapy. In some embodiments, this disclosure provides a method of treating a subject with cancer who has a corresponding need, the method comprising administering to the subject a therapeutically effective amount of the compounds herein (e.g., compound 1), wherein the cancer is platinum-sensitive, platinum-resistant, or platinum-refractory. In some embodiments, this disclosure provides a method of treating cancer in a subject with appropriate need, the method comprising administering to the subject a therapeutically effective amount of the compound herein (e.g., compound 1), wherein the subject has previously received platinum-based therapy for cancer, and that platinum-based therapy is ineffective in treating the cancer. In some embodiments, this disclosure provides a method of treating cancer in a subject with appropriate need, the method comprising administering to the subject a therapeutically effective amount of the compound herein (e.g., compound 1), at least in part based on the subject's prior receipt of platinum-based therapy for cancer, and the platinum-based therapy being ineffective in treating the cancer.

[0056] The compounds disclosed herein In some embodiments, the compounds of this disclosure comprise a substituted heterocyclic group, wherein the compound binds to the mutant p53 protein and increases the wild-type p53 activity of the mutant protein. In some embodiments, the compounds of this disclosure comprise a heterocyclic group containing a halogen substituent, wherein the compound binds to the mutant p53 protein and increases the wild-type p53 activity of the mutant protein. In some embodiments, the compound further comprises an indole group. In some embodiments, the indole group has a 2,2,2-trifluoroethyl substituent at the 1-position of the indole group.

[0057] In some embodiments, the indole group has a propargyl substituent at the 2-position. In some embodiments, the propargyl substituent is attached to the indole group via the sp carbon atom of the propargyl substituent. In some embodiments, the propargyl substituent is attached to the nitrogen atom of the aniline group via the methylene group of the propargyl substituent. In some embodiments, the indole group contains an amino substituent at the 4-position. In some embodiments, the amino substituent is attached to a heterocyclic group. In some embodiments, the heterocyclic group is a piperidine group. In some embodiments, the halogen substituent is a fluorine group. In some embodiments, the halogen substituent is a chlorine group. In some embodiments, the oral bioavailability of the compound is at least about 50% greater than that of similar compounds lacking a halogen substituent at the heterocyclic group.

[0058] In some implementations, the compound is Or its pharmaceutically acceptable salt.

[0059] In some embodiments, the compound is N-((3S,4R)-3-fluoro-1-methylpiperidin-4-yl)-2-(3-((2-methoxy-4-(methylsulfonyl)phenyl)amino)prop-1-yn-1-yl)-1-(2,2,2-trifluoroethyl)-1H-indole-4-amine or a pharmaceutically acceptable salt thereof.

[0060] In some implementations, the compound is (Compound 1) or a pharmaceutically acceptable salt thereof.

[0061] Or its pharmaceutically acceptable salt.

[0062] In some embodiments, the compound is 4-((3-(4-(((3S,4R)-3-fluoro-1-methylpiperidin-4-yl)amino)-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl)amino)-3-methoxy-N-methylbenzamide or a pharmaceutically acceptable salt thereof.

[0063] Suitable p53 activating compounds of this disclosure may also include, for example, the p53 activating compounds provided in US10138219B2, US11814373B2, US11807644B2, US11963953B2, US20230024905A1, US20230049952A1, PCT / US2024 / 046050 and PCT / US2024 / 046043, each of which is incorporated herein by reference in its entirety.

[0064] The compounds of the present invention may include all of their stereoisomers, enantiomers, diastereomers, mixtures, racemates, atropisomers and tautomers.

[0065] As used herein, the term "hydrogen", the atomic symbol "H", the implicit hydrogen atoms in chemical structures, and the hydrogen atoms covered by specific and general chemical group terms shall be understood to include all atoms with atomic number 1, including 1 H (i.e., "protium"), 2 H (i.e., "deuterium" or "D") and 3 H (i.e., "tritium"). For example, the terms "alkoxy" and "methoxy" shall be understood to include the group -OCD3, and the term "C1-C6 alkyl" shall include the term "C1-C6 deuterated alkyl". Similarly, all references to a given atomic element shall be understood to include all natural and unnatural isotopes of that element.

[0066] In certain embodiments, some or all of the 1 H atoms of the compounds disclosed herein are 2H atom substitution. Synthetic methods for deuterium-containing compounds are known in the art and include the following synthetic methods by way of non-limiting example only. Such deuterated forms can be prepared by procedures described in U.S. Patent Nos. 5,846,514 and 6,334,997. As described in U.S. Patent Nos. 5,846,514 and 6,334,997, deuteration can improve metabolic stability and / or potency, thereby increasing the duration of drug action. Deuterated compounds are synthesized using various methods such as those described in the following literature: Dean, Dennis C.; Recent Advances in the Synthesis and Applications of Radiolabeled Compounds for Drug Discovery and Development. [In: Curr., Pharm. Des., 2000; 6(10)] 2000, p. 110; George W.; Varma, Rajender S. The Synthesis of Radiolabeled Compounds via Organometallic Intermediates, Tetrahedron, 1989, 45(21), 6601-21; and Evans, E. Anthony. Synthesis of radiolabeled compounds, J. Radioanal. Chem., 1981, 64(1-2), 9-32. Deuterated starting materials are readily available and subjected to the synthetic methods described herein to provide the synthesis of deuterated compounds. Deuterium-containing reagents and building blocks are commercially available from chemical suppliers such as Aldrich Chemical Co.

[0067] In some embodiments, compositions comprising compounds disclosed herein can be prepared, whereby a certain proportion of atoms at a given position have a specific isotope, and said proportion differs from the natural abundance of said isotope. In some embodiments, this disclosure provides compositions comprising a molecular population, wherein each molecule has the structure of a compound (e.g., compound 1) provided herein, wherein the mass of the molecular population is at least 1 μg, and wherein at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 50%, at least 75%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, at least 99.9%, or at least 99.99% of the molecules in the molecular population each independently comprises a deuterium atom.

[0068] In some embodiments, this disclosure provides compositions comprising a molecular population of at least 1 μg, wherein each molecule has the structure of a compound (e.g., compound 1) provided herein, wherein if a mass spectrometry experiment is obtained of the population, the mass spectrometry experiment determines that at least 10% of the molecules in the molecular population independently contain deuterium atoms.

[0069] Any compound described herein can be purified. The compounds described herein can be at least 1% pure, at least 2% pure, at least 3% pure, at least 4% pure, at least 5% pure, at least 6% pure, at least 7% pure, at least 8% pure, at least 9% pure, at least 10% pure, at least 11% pure, at least 12% pure, at least 13% pure, at least 14% pure, at least 15% pure, at least 16% pure, at least 17% pure, at least 18% pure, at least 19% pure, at least 20% pure, at least 21% pure, at least 22% pure, at least 23% pure, at least 24% pure, at least 25% pure, at least 26% pure, at least 27% pure, at least 28% pure. % pure, at least 29% pure, at least 30% pure, at least 31% pure, at least 32% pure, at least 33% pure, at least 34% pure, at least 35% pure, at least 36% pure, at least 37% pure, at least 38% pure, at least 39% pure, at least 40% pure, at least 41% pure, at least 42% pure, at least 43% pure, at least 44% pure, at least 45% pure, at least 46% pure, at least 47% pure, at least 48% pure, at least 49% pure, at least 50% pure, at least 51% pure, at least 52% pure, at least 53% pure, at least 54% pure, at least 55% pure, to At least 56% pure, at least 57% pure, at least 58% pure, at least 59% pure, at least 60% pure, at least 61% pure, at least 62% pure, at least 63% pure, at least 64% pure, at least 65% pure, at least 66% pure, at least 67% pure, at least 68% pure, at least 69% pure, at least 70% pure, at least 71% pure, at least 72% pure, at least 73% pure, at least 74% pure, at least 75% pure, at least 76% pure, at least 77% pure, at least 78% pure, at least 79% pure, at least 80% pure, at least 81% pure, at least 82% pure, at least 83% pure Pure, at least 84% pure, at least 85% pure, at least 86% pure, at least 87% pure, at least 88% pure, at least 89% pure, at least 90% pure, at least 91% pure, at least 92% pure, at least 93% pure, at least 94% pure, at least 95% pure, at least 96% pure, at least 97% pure, at least 98% pure, at least 99% pure, at least 99% pure, at least 99.1% pure, at least 99.2% pure, at least 99.3% pure, at least 99.4% pure, at least 99.5% pure, at least 99.6% pure, at least 99.7% pure, at least 99.8% pure, or at least 99.9% pure.

[0070] Treatment In some embodiments, the compounds of this disclosure can be used to treat cancer in a subject. The compounds of this disclosure can, for example, slow the proliferation of cancer cell lines or kill cancer cells.Non-limiting examples of cancers that can be treated with the compounds disclosed herein include: acute lymphoblastic leukemia, acute myeloid leukemia, adrenocortical carcinoma, AIDS-related cancers, AIDS-related lymphomas, anal cancer, appendiceal cancer, astrocytoma, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brain tumors (e.g., cerebellar astrocytoma, astrocytoma / malignant glioma, ependymoma, medulloblastoma, supratentorial primitive neuroectodermal tumors, visual pathway and hypothalamic gliomas), breast cancer, bronchial adenoma, Burkitt lymphoma. Lymphoma, cancers of unknown origin, central nervous system lymphoma, cerebellar astrocytoma, cervical cancer, childhood cancer, chronic lymphocytic leukemia, chronic myeloid leukemia, chronic myelodysplastic disorder, colon cancer, cutaneous T-cell lymphoma, desmoplastic small round cell tumor, endometrial cancer, ependymoma, esophageal cancer, Ewing's sarcoma, germ cell tumors, gallbladder cancer, gastric cancer, gastrointestinal carcinoid tumors, gastrointestinal stromal tumors, glioma. Hairy cell leukemia, head and neck cancer, heart cancer, hepatocellular carcinoma, Hodgkin's lymphoma, hypopharyngeal cancer, intraocular melanoma, islet cell carcinoma, Kaposi's sarcoma, kidney cancer, laryngeal cancer, lip and oral cancer, liposarcoma, liver cancer, lung cancer (such as non-small cell lung cancer and small cell lung cancer), lymphoma, leukemia, macroglobulinemia, malignant fibrous histiocytoma, medulloblastoma, melanoma, mesothelioma, metastatic squamous neck cancer of unknown primary origin. Occult primary), oral cancer, multiple endocrine neoplasia syndrome, myelodysplastic syndrome, myeloid leukemia, nasal cavity and paranasal sinus cancer, nasopharyngeal carcinoma, neuroblastoma, non-Hodgkin lymphoma, non-small cell lung cancer, oral cancer, oropharyngeal cancer, osteosarcoma, ovarian cancer, high-grade serous ovarian cancer, ovarian cancer with homologous recombination defect, ovarian epithelial cancer, ovarian germ cell tumor, pancreatic cancer, islet cell pancreatic cancer, paranasal sinus and nasal cavity cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pheochromocytoma, pineal astrocytoma, pineal germ cell tumor, pituitary adenoma Pleural pulmonary blastoma, plasmacytoma formation, primary central nervous system lymphoma, prostate cancer, rectal cancer, renal cell carcinoma, transitional cell carcinoma of the renal pelvis and ureter, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sarcoma, skin cancer, Merkel cell skin cancer, small intestine cancer, soft tissue sarcoma, squamous cell carcinoma, gastric cancer, T-cell lymphoma, pharyngeal cancer, thymoma, thymic carcinoma, thyroid cancer, trophoblastic tumor (pregnancy-related), cancer of unknown primary site, urethral cancer, uterine sarcoma, vaginal cancer, vulvar cancer, Waldenström macroglobulinemia, and Wilms tumor.

[0071] In some embodiments, the compounds of this disclosure can be used to treat breast cancer in a subject. In some embodiments, the breast cancer is human epidermal growth receptor 2 positive (HER2) breast cancer. In some embodiments, the breast cancer is hormone receptor negative (HR1) breast cancer. In some embodiments, the breast cancer is human epidermal growth receptor 2 negative (HER2) breast cancer. In some embodiments, the breast cancer is both hormone receptor positive (HR1) and human epidermal growth receptor 2 positive (HER2) breast cancer. In some embodiments, the breast cancer is both hormone receptor negative (HR1) and human epidermal growth receptor 2 negative (HER2) breast cancer. In some embodiments, the breast cancer is both hormone receptor negative (HR1) and human epidermal growth receptor 2 positive (HER2) breast cancer.

[0072] In some embodiments, this disclosure provides a method for treating triple-negative breast cancer (TNBC), the method comprising administering a therapeutically effective amount of the compound of this disclosure to a subject with appropriate need. Triple-negative breast cancer (TNBC) refers to any breast cancer that does not express the genes for estrogen receptor (ER), progesterone receptor (PR), or Her2 / neu.

[0073] In some embodiments, this disclosure provides a method for treating cancer-related edema, the method comprising administering a therapeutically effective amount of a compound of this disclosure to a subject with appropriate need. In some embodiments, the cancer-related edema is associated with breast cancer. In some embodiments, the cancer-related edema also includes limitation of limb or finger / toe movement. In some embodiments, the cancer-related edema also includes limitation of arm movement. In some embodiments, the cancer-related edema is lymphedema.

[0074] In some embodiments, this disclosure provides a method for improving cancer-related edema in a cancer subject with a corresponding need, the method comprising administering a therapeutically effective amount of a compound of this disclosure to the cancer subject, wherein the cancer subject has cancer-related edema prior to administration and experiences improvement in cancer-related edema after administration (e.g., after a course of treatment with the compound of this disclosure). In some embodiments, the cancer-related edema is lymphedema. In some embodiments, the cancer subject has breast cancer in the breast. In some embodiments, the cancer-related edema is in the breast with breast cancer. In some embodiments, the cancer-related edema is in a limb. In some embodiments, the limb is the arm. In some embodiments, the limb is the arm, wherein the breast is on the same side as the arm. In some embodiments, the cancer-related edema is in the fingers / toes. In some embodiments, the fingers / toes are the fingers. In some embodiments, the fingers / toes are the fingers, wherein the breast is on the same side as the fingers.

[0075] In some embodiments, this disclosure provides a method for improving limb mobility in a cancer subject with a corresponding need, the method comprising administering to the cancer subject a therapeutically effective amount of a compound of this disclosure, wherein the cancer subject had limited limb mobility prior to administration and experienced improvement in limb mobility after administration (e.g., after a course of treatment with a compound of this disclosure). In some embodiments, the limb is an arm. In some embodiments, the cancer subject has breast cancer. In some embodiments, the limb is an arm, wherein the breast cancer is located in the breast on the same side as the arm.

[0076] In some embodiments, this disclosure provides a method comprising: a) administering a compound of this disclosure to a subject requiring cancer therapy; b) observing an improvement in limb mobility in the subject following administration; and c) further administration of the compound of this disclosure based at least in part on the observed improvement in limb mobility in the subject following administration. In some embodiments, the subject has breast cancer, and the cancer therapy is a breast cancer therapy. In some embodiments, the method further includes observing that the subject exhibits suppressed limb mobility prior to the administration. In some embodiments, the limb is the arm. In some embodiments, the cancer subject has breast cancer. In some embodiments, the limb is the arm, wherein the breast cancer is located in the breast on the same side as the arm.

[0077] In some embodiments, this disclosure provides a method for improving finger / toe mobility in a cancer subject with a corresponding need, the method comprising administering a therapeutically effective amount of a compound of this disclosure to the cancer subject, wherein the cancer subject has limited finger / toe mobility prior to administration and experiences improvement in finger / toe mobility after administration (e.g., after a course of treatment with a compound of this disclosure). In some embodiments, the finger / toe is a finger. In some embodiments, the cancer subject has breast cancer. In some embodiments, the finger / toe is a finger, wherein the breast cancer is located in the breast on the same side as the finger.

[0078] In some embodiments, this disclosure provides a method comprising: a) administering a compound of this disclosure to a subject requiring cancer therapy; b) observing an improvement in finger / toe mobility in the subject following administration; and c) further administration of the compound of this disclosure based at least in part on the observed improvement in finger / toe mobility in the subject following administration. In some embodiments, the subject has breast cancer, and the cancer therapy is a breast cancer therapy. In some embodiments, the method further includes observing inhibition of finger / toe mobility in the subject prior to said administration. In some embodiments, finger / toe refers to a finger. In some embodiments, the cancer subject has breast cancer. In some embodiments, finger / toe refers to a finger, wherein the breast cancer is located in the breast on the same side as the finger.

[0079] In some embodiments, the compounds of this disclosure exhibit non-lethal toxicity.

[0080] Pharmacokinetic and pharmacodynamic properties Pharmacokinetic and pharmacodynamic data can be obtained through a variety of experimental techniques. The appropriate pharmacokinetic and pharmacodynamic profile component describing a particular composition can vary due to variations in drug metabolism in human subjects. Pharmacokinetic and pharmacodynamic profiles can be determined based on the determination of mean parameters in a subject group. A subject group includes any reasonable number of subjects suitable for determining representative mean values, such as 5, 10, 15, 20, 25, 30, 35, or more subjects. The mean is determined by calculating the average of the measurements of each parameter across all subjects. As described herein, the dose can be adjusted to obtain a desired pharmacokinetic or pharmacodynamic profile, such as a desired or effective hematologic profile.

[0081] Pharmacodynamic parameters can be any parameters suitable for describing the compositions of the present invention. For example, pharmacodynamic profiles can be obtained at the following times after administration: for example, about 0 minutes, about 1 minute, about 2 minutes, about 3 minutes, about 4 minutes, about 5 minutes, about 6 minutes, about 7 minutes, about 8 minutes, about 9 minutes, about 10 minutes, about 11 minutes, about 12 minutes, about 13 minutes, about 14 minutes, about 15 minutes, about 16 minutes, about 17 minutes, about 18 minutes, about 19 minutes, about 20 minutes, about 21 minutes, about 22 minutes, about 23 minutes, about 24 minutes, about 25 minutes, about 26 minutes. Approximately 27 minutes, approximately 28 minutes, approximately 29 minutes, approximately 30 minutes, approximately 31 minutes, approximately 32 minutes, approximately 33 minutes, approximately 34 minutes, approximately 35 minutes, approximately 36 minutes, approximately 37 minutes, approximately 38 minutes, approximately 39 minutes, approximately 40 minutes, approximately 41 minutes, approximately 42 minutes, approximately 43 minutes, approximately 44 minutes, approximately 45 minutes, approximately 46 minutes, approximately 47 minutes, approximately 48 minutes, approximately 49 minutes, approximately 50 minutes, approximately 51 minutes, approximately 52 minutes, approximately 53 minutes, approximately 54 minutes, approximately 55 minutes. Approximately 56 minutes, approximately 57 minutes, approximately 58 minutes, approximately 59 minutes, approximately 60 minutes, approximately 0 hours, approximately 0.5 hours, approximately 1 hour, approximately 1.5 hours, approximately 2 hours, approximately 2.5 hours, approximately 3 hours, approximately 3.5 hours, approximately 4 hours, approximately 4.5 hours, approximately 5 hours, approximately 5.5 hours, approximately 6 hours, approximately 6.5 hours, approximately 7 hours, approximately 7.5 hours, approximately 8 hours, approximately 8.5 hours, approximately 9 hours, approximately 9.5 hours, approximately 10 hours, approximately 10.5 hours, approximately 11 hours, approximately 11.5 hours Approximately 12 hours, approximately 12.5 hours, approximately 13 hours, approximately 13.5 hours, approximately 14 hours, approximately 14.5 hours, approximately 15 hours, approximately 15.5 hours, approximately 16 hours, approximately 16.5 hours, approximately 17 hours, approximately 17.5 hours, approximately 18 hours, approximately 18.5 hours, approximately 19 hours, approximately 19.5 hours, approximately 20 hours, approximately 20.5 hours, approximately 21 hours, approximately 21.5 hours, approximately 22 hours, approximately 22.5 hours, approximately 23 hours, approximately 23.5 hours, or approximately 24 hours.

[0082] Pharmacokinetic parameters can be any parameters suitable for describing a compound. Non-limiting examples of pharmacodynamic and pharmacokinetic parameters that can be calculated for a compound administered using the methods of this invention include: Non-limiting examples of pharmacokinetic parameters that can be used to determine the effect of treating a subject with the compositions of this disclosure include: a terminal elimination rate constant (λz); and a pharmacokinetic parameter exceeding t last AUC obtained by extrapolation 0-inf Percentage (AUC) %extrapThe area under the concentration-time curve (AUC) from before administration (time 0) to 24 hours later. 0-24 The area under the concentration-time curve (AUC) from before administration (time 0) to 96 hours later. 0-96 The area under the concentration-time curve (AUC) extrapolated from before administration (time 0) to infinity. 0-inf From before administration (time 0) to t last Area under the concentration-time curve (AUC) 0-last ); Apparent total body clearance (CL / F); Observed maximum plasma concentration (C max ); Apparent terminal elimination half-life (t½); Time before first measurable (non-zero) concentration (t lag The final time for quantifying the concentration (t) last ); corresponding to C max Time of occurrence (T) max ); and apparent distribution volume (Vz / F). C max For example, not less than about 1 ng / mL; not less than about 5 ng / mL; not less than about 10 ng / mL; not less than about 15 ng / mL; not less than about 20 ng / mL; not less than about 25 ng / mL; not less than about 50 ng / mL; not less than about 75 ng / mL; not less than about 100 ng / mL; not less than about 200 ng / mL; not less than about 300 ng / mL; not less than about 400 ng / mL; not less than about 500 ng / mL; not less than about 600 ng / mL; not less than about 700 ng / mL; not less than about 800 ng / mL; not less than about 900 ng / mL; not less than about 1000 ng / mL; not less than about 1250 ng / mL; not less than about 1500 ng / mL; not less than about 1750 ng / mL; not less than about 2000 ng / mL. ng / mL; or any other C ng / mL suitable for describing the pharmacokinetic profile of the compounds described herein. max C maxIt can be, for example, about 1 ng / mL to about 5,000 ng / mL; about 1 ng / mL to about 4,500 ng / mL; about 1 ng / mL to about 4,000 ng / mL; about 1 ng / mL to about 3,500 ng / mL; about 1 ng / mL to about 3,000 ng / mL; about 1 ng / mL to about 2,500 ng / mL; about 1 ng / mL to about 2,000 ng / mL; about 1 ng / mL to about 1,500 ng / mL; about 1 ng / mL to about 1,000 ng / mL; about 1 ng / mL to about 900 ng / mL; about 1 ng / mL to about 800 ng / mL; about 1 ng / mL to about 700 ng / mL; about 1 ng / mL to about 600 ng / mL; about 1 ng / mL to about 500 ng / mL; about 1 ng / mL to about 450 ng / mL. ng / mL; about 1 ng / mL to about 400 ng / mL; about 1 ng / mL to about 350 ng / mL; about 1 ng / mL to about 300 ng / mL; about 1 ng / mL to about 250 ng / mL; about 1 ng / mL to about 200 ng / mL; about 1 ng / mL to about 150 ng / mL; about 1 ng / mL to about 125 ng / mL; about 1 ng / mL to about 100 ng / mL; about 1 ng / mL to about 90 ng / mL; about 1 ng / mL to about 80 ng / mL; about 1 ng / mL to about 70 ng / mL; about 1 ng / mL to about 60 ng / mL; about 1 ng / mL to about 50 ng / mL; about 1 ng / mL to about 40 ng / mL; about 1 ng / mL to about 30 ng / mL; about 1 ng / mL to about 20 ng / mL; about 1 ng / mL to about 10 ng / mL; about 1 ng / mL to about 5 ng / mL; about 10 ng / mL to about 4,000 ng / mL; about 10 ng / mL to about 3,000 ng / mL; about 10 ng / mL to about 2,000 ng / mL; about 10 ng / mL to about 1,500 ng / mL; about 10 ng / mL to about 1,000 ng / mL; about 10 ng / mL to about 900 ng / mL; about 10 ng / mL to about 800 ng / mL; about 10 ng / mL to about 700 ng / mL; about 10 ng / mL to about 600 ng / mL; about 10 ng / mL to about 500 ng / mL; about 10 ng / mL to about 400 ng / mL; about 10 ng / mL to about 300 ng / mL; about 10 ng / mL to about 200 ng / mL; about 10 ng / mL to about 100 ng / mL; about 10 ng / mL to about 50 ng / mL;Approximately 25 ng / mL to approximately 500 ng / mL; approximately 25 ng / mL to approximately 100 ng / mL; approximately 50 ng / mL to approximately 500 ng / mL; approximately 50 ng / mL to approximately 100 ng / mL; approximately 100 ng / mL to approximately 500 ng / mL; approximately 100 ng / mL to approximately 400 ng / mL; approximately 100 ng / mL to approximately 300 ng / mL; or approximately 100 ng / mL to approximately 200 ng / mL.

[0083] In some implementation schemes, C max The concentration is approximately 8 mg / mL to approximately 15 mg / mL. In some embodiments, C max The concentration is approximately 8 mg / mL to approximately 12 mg / mL. In some embodiments, C max The concentration is approximately 7 mg / mL to approximately 11 mg / mL. In some embodiments, C max The concentration is approximately 12 mg / mL to approximately 28 mg / mL. In some embodiments, C max The concentration is approximately 3 mg / mL to approximately 20 mg / mL. In some embodiments, C max The concentration is approximately 7 mg / mL to approximately 20 mg / mL.

[0084] In some embodiments, the C-value of the administered compound is compared with that of the subject who has not consumed food for a certain period of time prior to administration of the compound of this disclosure. max In contrast, in the method disclosed herein, where the subject has consumed food within a certain timeframe prior to administration of the compound of this disclosure, C max Increase. In some embodiments, the C of the compounds disclosed herein is... max It increased by approximately 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85% or more.

[0085] In some implementations, C of fasting subjects max In comparison, the C of the compound disclosed in this article in the subjects who ingested it... max This increased by approximately 28%. In some implementations, compared to the C of fasting subjects... max In comparison, the C of the compound disclosed in this article in the subjects who ingested it... max This increased by approximately 35%. In some implementations, compared to the C of fasting subjects... max In comparison, the C of the compound disclosed in this article in the subjects who ingested it... max It increased by approximately 84%.

[0086] The T of the compounds described in this article maxThis can be, for example, no more than about 0.5 hours, no more than about 1 hour, no more than about 1.5 hours, no more than about 2 hours, no more than about 2.5 hours, no more than about 3 hours, no more than about 3.5 hours, no more than about 4 hours, no more than about 4.5 hours, no more than about 5 hours, or any other T suitable for describing the pharmacokinetic profile of the compounds described herein. max T max It can be, for example, about 0.1 hours to about 24 hours; about 0.1 hours to about 0.5 hours; about 0.5 hours to about 1 hour; about 1 hour to about 1.5 hours; about 1.5 hours to about 2 hours; about 2 hours to about 2.5 hours; about 2.5 hours to about 3 hours; about 3 hours to about 3.5 hours; about 3.5 hours to about 4 hours; about 4 hours to about 4.5 hours; about 4.5 hours to about 5 hours; about 5 hours to about 5.5 hours; about 5.5 hours to about 6 hours. Approximately 6 hours to approximately 6.5 hours; approximately 6.5 hours to approximately 7 hours; approximately 7 hours to approximately 7.5 hours; approximately 7.5 hours to approximately 8 hours; approximately 8 hours to approximately 8.5 hours; approximately 8.5 hours to approximately 9 hours; approximately 9 hours to approximately 9.5 hours; approximately 9.5 hours to approximately 10 hours; approximately 10 hours to approximately 10.5 hours; approximately 10.5 hours to approximately 11 hours; approximately 11 hours to approximately 11.5 hours; approximately 11.5 hours to approximately 12 hours; approximately 12 hours to approximately 12. 5 hours; approximately 12.5 hours to approximately 13 hours; approximately 13 hours to approximately 13.5 hours; approximately 13.5 hours to approximately 14 hours; approximately 14 hours to approximately 14.5 hours; approximately 14.5 hours to approximately 15 hours; approximately 15 hours to approximately 15.5 hours; approximately 15.5 hours to approximately 16 hours; approximately 16 hours to approximately 16.5 hours; approximately 16.5 hours to approximately 17 hours; approximately 17 hours to approximately 17.5 hours; approximately 17.5 hours to approximately 18 hours; approximately 18 hours From about 18.5 hours; from about 18.5 hours to about 19 hours; from about 19 hours to about 19.5 hours; from about 19.5 hours to about 20 hours; from about 20 hours to about 20.5 hours; from about 20.5 hours to about 21 hours; from about 21 hours to about 21.5 hours; from about 21.5 hours to about 22 hours; from about 22 hours to about 22.5 hours; from about 22.5 hours to about 23 hours; from about 23 hours to about 23.5 hours; or from about 23.5 hours to about 24 hours. In some embodiments, the T of the compounds of this disclosure max For approximately 2 hours. In some embodiments, the T of the compounds disclosed herein... max For approximately 4 hours. In some embodiments, the T of the compounds disclosed herein... max For approximately 6 hours. In some embodiments, the T of the compounds disclosed herein... max It takes about 8 hours.

[0087] In some implementations, Tmax The duration is approximately 1.5 hours to approximately 5 hours. In some implementations, T max The duration is approximately 1.5 hours to approximately 2.5 hours. In some implementations, T max The duration is approximately 2 hours to approximately 8 hours. In some implementations, T max The duration is approximately 1 hour to approximately 6 hours. In some implementations, T max The duration is approximately 2 to 5 hours. In some implementations, T max It takes approximately 2 hours. In some implementations, T max It takes approximately 3 hours. In some implementations, T max It takes about 4 hours.

[0088] In some embodiments, the T value of the administered compound is compared with that of the subject who has not consumed food for a certain period of time prior to administration of the compound of this disclosure. max In contrast, in the method disclosed herein, where the subject has consumed food within a certain timeframe prior to administration of the compound of this disclosure, T max Increase. In some embodiments, the T of the compounds disclosed herein max It increased by approximately 15%, approximately 20%, approximately 25%, approximately 30%, approximately 35%, approximately 40%, approximately 45%, approximately 50% or more.

[0089] AUC of the compounds described in this article (0-inf) or AUC (last)For example, not less than about 1 ng·hr / mL, not less than about 5 ng·hr / mL, not less than about 10 ng·hr / mL, not less than about 20 ng·hr / mL, not less than about 30 ng·hr / mL, not less than about 40 ng·hr / mL, not less than about 50 ng·hr / mL, not less than about 100 ng·hr / mL, not less than about 150 ng·hr / mL, not less than about 200 ng·hr / mL, not less than about 250 ng·hr / mL, not less than about 300 ng·hr / mL, not less than about 350 ng·hr / mL, not less than about 400 ng·hr / mL, not less than about 450 ng·hr / mL, not less than about 500 ng·hr / mL, not less than about 600 ng·hr / mL, not less than about 700 ng·hr / mL, not less than about 800 ng·hr / mL, not less than about 900 ng·hr / mL. Not less than about 1000 ng·hr / mL, not less than about 1250 ng·hr / mL, not less than about 1500 ng·hr / mL, not less than about 1750 ng·hr / mL, not less than about 2000 ng·hr / mL, not less than about 2500 ng·hr / mL, not less than about 3000 ng·hr / mL, not less than about 3500 ng·hr / mL, not less than about 4000 ng·hr / mL, not less than about 5000 ng·hr / mL, not less than about 6000 ng·hr / mL, not less than about 7000 ng·hr / mL, not less than about 8000 ng·hr / mL, not less than about 9000 ng·hr / mL, not less than about 10000 ng·hr / mL, or any other AUC suitable for describing the pharmacokinetic profile of the compounds described herein. (0-inf) or AUC (last) In some embodiments, the AUC of the compounds described herein (0-inf) or AUC (last)For example, not less than about 10,000 ng·hr / mL, not less than about 11,000 ng·hr / mL, not less than about 12,000 ng·hr / mL, not less than about 13,000 ng·hr / mL, not less than about 14,000 ng·hr / mL, not less than about 15,000 ng·hr / mL, not less than about 16,000 ng·hr / mL, not less than about 17,000 ng·hr / mL, not less than about 18,000 ng·hr / mL, not less than about 19,000 ng·hr / mL, not less than about 20,000 ng·hr / mL, not less than about 21,000 ng·hr / mL, not less than about 22,000 ng·hr / mL, not less than about 23,000 ng·hr / mL, not less than about 24,000 ng·hr / mL. ng·hr / mL or not less than about 25,000 ng·hr / mL.

[0090] AUC of the compound (0-inf) or AUC (last)It can be, for example, about 1 ng·hr / mL to about 10,000 ng·hr / mL; about 1 ng·hr / mL to about 10 ng·hr / mL; about 10 ng·hr / mL to about 25 ng·hr / mL; about 25 ng·hr / mL to about 50 ng·hr / mL; about 50 ng·hr / mL to about 100 ng·hr / mL; about 100 ng·hr / mL to about 200 ng·hr / mL; about 200 ng·hr / mL to about 300 ng·hr / mL; about 300 ng·hr / mL to about 400 ng·hr / mL; about 400 ng·hr / mL to about 500 ng·hr / mL; about 500 ng·hr / mL to about 600 ng·hr / mL; about 600 ng·hr / mL to about 700 ng·hr / mL; about 700 ng·hr / mL to about 800 ng·hr / mL. ng·hr / mL; about 800 ng·hr / mL to about 900 ng·hr / mL; about 900 ng·hr / mL to about 1,000 ng·hr / mL; about 1,000 ng·hr / mL to about 1,250 ng·hr / mL; about 1,250 ng·hr / mL to about 1,500 ng·hr / mL; about 1,500 ng·hr / mL to about 1,750 ng·hr / mL; about 1,750 ng·hr / mL to about 2,000 ng·hr / mL; about 2,000 ng·hr / mL to about 2,500 ng·hr / mL; about 2,500 ng·hr / mL to about 3,000 ng·hr / mL; about 3,000 ng·hr / mL to about 3,500 ng·hr / mL; about 3,500 ng·hr / mL to about 4,000 ng·hr / mL ng·hr / mL; about 4,000 ng·hr / mL to about 4,500 ng·hr / mL; about 4,500 ng·hr / mL to about 5,000 ng·hr / mL; about 5,000 ng·hr / mL to about 5,500 ng·hr / mL; about 5,500 ng·hr / mL to about 6,000 ng·hr / mL; about 6,000 ng·hr / mL to about 6,500 ng·hr / mL; about 6,500 ng·hr / mL to about 7,000 ng·hr / mL; about 7,000 ng·hr / mL to about 7,500 ng·hr / mL; about 7,500 ng·hr / mL to about 8,000 ng·hr / mL; about 8,000 ng·hr / mL to about 8,500 ng·hr / mL; about 8,500 ng·hr / mL to about 9,000 ng·hr / mL ng·hr / mL; approximately 9,000 ng·hr / mL to approximately 9,500 ng·hr / mL;Or about 9,500 ng·hr / mL to about 10,000 ng·hr / mL. In some embodiments, the AUC of the compounds described herein; (0-inf) or AUC (last) It can be, for example, about 10,000 ng·hr / mL, about 11,000 ng·hr / mL, about 12,000 ng·hr / mL, about 13,000 ng·hr / mL, about 14,000 ng·hr / mL, about 15,000 ng·hr / mL, about 16,000 ng·hr / mL, about 17,000 ng·hr / mL, about 18,000 ng·hr / mL, about 19,000 ng·hr / mL, about 20,000 ng·hr / mL, about 21,000 ng·hr / mL, about 22,000 ng·hr / mL, about 23,000 ng·hr / mL, about 24,000 ng·hr / mL, or about 25,000 ng·hr / mL.

[0091] The area under the concentration-time curve (AUC) from before administration (time 0) to 24 hours. 0-24 () can be, for example, about 90,000 ng·hr / mL to about 500,000 ng·hr / mL, about 100,000 ng·hr / mL to about 500,000 ng·hr / mL, about 150,000 ng·hr / mL to about 500,000 ng·hr / mL, about 200,000 ng·hr / mL to about 500,000 ng·hr / mL, about 100,000 ng·hr / mL to about 400,000 ng·hr / mL, about 100,000 ng·hr / mL to about 300,000 ng·hr / mL, about 100,000 ng·hr / mL to about 250,000 ng·hr / mL, about 100,000 ng·hr / mL to about 200,000 ng·hr / mL, about 15 ... From approximately 150,000 ng·hr / mL to approximately 200,000 ng·hr / mL. In some embodiments, the AUC is measured in subjects receiving a therapeutically effective amount of the compound once daily. 0-24 In some implementation schemes, AUC is measured in subjects on days 10, 11, 12, 13, 14, or 15 of the treatment regimen. 0-24 The treatment regimen includes administering a therapeutically effective amount of the compound once daily. In some embodiments, the AUC is measured in subjects on day fifteen of the treatment regimen. 0-24In some embodiments, the therapeutically effective dose is 2000 mg. In some embodiments, the therapeutically effective dose is 3000 mg. In some embodiments, the therapeutically effective dose is administered orally. In some embodiments, the therapeutically effective dose is administered with food.

[0092] In some embodiments, this disclosure provides pharmaceutical compositions comprising the p53-activating compound of this disclosure, wherein, following 15 once-daily administrations of a pharmaceutical composition comprising a therapeutically effective amount of the compound, an AUC of approximately 90,000 ng·hr / mL to approximately 500,000 ng·hr / mL of the compound is achieved in subjects. 0-24 In some embodiments, the therapeutically effective dose is 2000 mg. In some embodiments, the therapeutically effective dose is 3000 mg. In some embodiments, the therapeutically effective dose is administered orally. In some embodiments, the therapeutically effective dose is administered with food.

[0093] In some embodiments, the food includes a high-fat diet. In some embodiments, the high-fat diet comprises at least 50% of the total calorie content of the diet as fat. In some embodiments, the high-fat diet comprises at least 500 kcal of fat. In some embodiments, the high-fat diet comprises at least 55 g of fat.

[0094] In some embodiments, the food comprises a moderate-fat diet. In some embodiments, a moderate-fat diet comprises approximately 25% to approximately 50% of the total calorie content of the diet as fat. In some embodiments, a moderate-fat diet comprises approximately 125 kcal to approximately 500 kcal of fat. In some embodiments, a moderate-fat diet comprises approximately 14 g to approximately 55 g of fat.

[0095] In some embodiments, the food includes a low-fat diet. In some embodiments, a low-fat diet comprises less than 25% of the total calorie content of the diet as fat. In some embodiments, a low-fat diet comprises less than about 125 kcal of fat from fat. In some embodiments, a low-fat diet comprises less than about 14 g of fat.

[0096] In some implementations, a high-fat diet contains approximately 150 calories from protein, approximately 250 calories from carbohydrates, and approximately 500 to 600 calories from fat.

[0097] In some embodiments, the food includes a high-calorie diet. In some embodiments, the high-calorie diet contains at least 800 calories. In some embodiments, the high-calorie diet contains approximately 800 calories to approximately 1000 calories.

[0098] In some embodiments, the food includes a high-fat, high-calorie diet. In some embodiments, a high-fat, high-calorie diet comprises at least 50% fat and approximately 800 to approximately 1000 calories of total calories.

[0099] In some embodiments, the AUC of the disclosed method is increased compared to the AUC of the administered compound in which the subject has consumed food for a certain period of time prior to administration of the compound of this disclosure. In some embodiments, the AUC of the disclosed compound is increased by about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, or more. In some embodiments, the AUC is AUC 0-inf In some implementations, AUC is AUC 0-last In some implementations, AUC is AUC 0-24 .

[0100] In some implementations, the AUC of the fasting subject (e.g., AUC) is compared with that of the fasting subject. 0-Inf AUC 0-last AUC 0-24 Compared to the AUC (e.g., AUC) of the compounds disclosed in this paper in subjects who ingested the compound, 0-Inf AUC 0-last AUC 0-24 The AUC of fasting subjects increased by approximately 40%. In some implementations, this is compared with the AUC of fasting subjects (e.g., AUC...). 0-Inf AUC 0-last AUC 0-24 Compared to the AUC (e.g., AUC) of the compounds disclosed in this paper in subjects who ingested the compound, 0-Inf AUC 0-last AUC 0-24 The AUC of fasting subjects increased by approximately 45%. In some implementations, this is compared with the AUC of fasting subjects (e.g., AUC...). 0-Inf AUC 0-last AUC 0-24 Compared to the AUC (e.g., AUC) of the compounds disclosed in this paper in subjects who ingested the compound, 0-Inf AUC 0-lastAUC 0-24 It increased by approximately 79%.

[0101] The apparent total plasma clearance (CL / F) of the compounds disclosed herein can be, for example, about 2 L / h, about 2.5 L / h, about 3 L / h, about 3.5 L / h, about 4 L / h, about 4.5 L / h, about 5 L / h, about 5.5 L / h, about 6 L / h, about 6.5 L / h, about 7 L / h, about 7.5 L / h, about 8 L / h, about 8.5 L / h, about 9 L / h, about 9.5 L / h, about 10 L / h, about 10.5 L / h, about 11 L / h, about 11.5 L / h, about 12 L / h, about 12.5 L / h, about 13 L / h, about 13.5 L / h, about 14 L / h, about 14.5 L / h, about 15 L / h, about 15.5 L / h, about 16 L / h, about 16.5 L / h, about 17 ... L / h, approximately 17.5 L / h, approximately 18 L / h, approximately 18.5 L / h, approximately 19 L / h, approximately 19.5 L / h, or approximately 20 L / h.

[0102] In some embodiments, the CL / F of the applied compound is reduced in methods disclosed herein where the subject has consumed food for a certain period of time prior to the administration of the compound of this disclosure, compared to the CL / F of the applied compound where the subject has not consumed food for a certain period of time prior to the administration of the compound of this disclosure. In some embodiments, the CL / F of the compound of this disclosure is reduced by about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, or more.

[0103] The apparent plasma distribution volume (Vz / F) of the compounds disclosed herein can be, for example, about 90 L, about 100 L, about 110 L, about 120 L, about 130 L, about 135 L, about 140 L, about 145 L, about 150 L, about 155 L, about 160 L, about 165 L, about 170 L, about 175 L, about 180 L, about 185 L, about 190 L, about 195 L, about 200 L, about 225 L, about 250 L, about 275 L, about 300 L, about 325 L, about 350 L, or about 400 L.

[0104] In some embodiments, the Vz / F is reduced in the methods disclosed herein where the subject has consumed food for a certain period of time prior to the administration of the compound of this disclosure, compared to the Vz / F of the administered compound where the subject has not consumed food for a certain period of time prior to the administration of the compound of this disclosure. In some embodiments, the Vz / F of the compounds disclosed herein is reduced by about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, or more.

[0105] The plasma concentrations of the compounds described herein may be, for example, not less than about 1 ng / mL, not less than about 5 ng / mL, not less than about 10 ng / mL, not less than about 15 ng / mL, not less than about 20 ng / mL, not less than about 25 ng / mL, not less than about 50 ng / mL, not less than about 75 ng / mL, not less than about 100 ng / mL, not less than about 150 ng / mL, not less than about 200 ng / mL, not less than about 300 ng / mL, not less than about 400 ng / mL, not less than about 500 ng / mL, not less than about 600 ng / mL, not less than about 700 ng / mL, not less than about 800 ng / mL, not less than about 900 ng / mL, not less than about 1000 ng / mL, not less than about 1200 ng / mL, or any other plasma concentration of the compounds described herein.Plasma concentrations can be, for example, from about 1 ng / mL to about 2,000 ng / mL; from about 1 ng / mL to about 5 ng / mL; from about 5 ng / mL to about 10 ng / mL; from about 10 ng / mL to about 25 ng / mL; from about 25 ng / mL to about 50 ng / mL; from about 50 ng / mL to about 75 ng / mL; from about 75 ng / mL to about 100 ng / mL; from about 100 ng / mL to about 150 ng / mL; from about 150 ng / mL to about 200 ng / mL; from about 200 ng / mL to about 250 ng / mL; from about 250 ng / mL to about 300 ng / mL; from about 300 ng / mL to about 350 ng / mL; from about 350 ng / mL to about 400 ng / mL; from about 400 ng / mL to about 450 ng / mL; from about 450 ng / mL to about 500 ng / mL. ng / mL; about 500 ng / mL to about 600 ng / mL; about 600 ng / mL to about 700 ng / mL; about 700 ng / mL to about 800 ng / mL; about 800 ng / mL to about 900 ng / mL; about 900 ng / mL to about 1,000 ng / mL; about 1,000 ng / mL to about 1,100 ng / mL; about 1,100 ng / mL to about 1,200 ng / mL; about 1,200 ng / mL to about 1,300 ng / mL; about 1,300 ng / mL to about 1,400 ng / mL; about 1,400 ng / mL to about 1,500 ng / mL; about 1,500 ng / mL to about 1,600 ng / mL; about 1,600 ng / mL to about 1,700 ng / mL; about 1,700 ng / mL From about 1,800 ng / mL; from about 1,800 ng / mL to about 1,900 ng / mL; or from about 1,900 ng / mL to about 2,000 ng / mL.

[0106] In some implementations, the plasma concentration may be about 2,500 ng / mL, about 3,000 ng / mL, about 3,500 ng / mL, about 4,000 ng / mL, about 4,500 ng / mL, about 5,000 ng / mL, about 5,500 ng / mL, about 6,000 ng / mL, about 6,500 ng / mL, about 7,000 ng / mL, about 7,500 ng / mL, about 8,000 ng / mL, about 8,500 ng / mL, about 9,000 ng / mL, about 9,500 ng / mL, or about 10,000 ng / mL. In some implementations, the plasma concentration may be about 10,000 ng / mL, about 15,000 ng / mL, about 20,000 ng / mL, about 25,000 ng / mL, about 30,000 ng / mL, about 35,000 ng / mL, about 40,000 ng / mL, about 45,000 ng / mL, about 50,000 ng / mL, about 55,000 ng / mL, about 60,000 ng / mL, about 65,000 ng / mL, about 70,000 ng / mL, or about 75,000 ng / mL.

[0107] Further non-limiting examples of pharmacokinetic parameters that can be used to determine the effect of treating a subject with the compositions of this disclosure include: the amount of drug excreted in urine without change (Ae), renal clearance (CLR), and fraction of drug excreted in urine (fe).

[0108] Pharmacodynamic parameters can be any parameters suitable for describing the compositions described in this disclosure. For example, a pharmacodynamic profile may show a decrease in the viability phenotype of tumor cells or a reduction in tumor size in tumor cell lines or xenograft studies, for example, about 24 hours, about 48 hours, about 72 hours, or 1 week.

[0109] Food effect In some embodiments, the compounds disclosed herein are administered within 30 minutes after food consumption. In some embodiments, the compounds disclosed herein are administered concurrently with food consumption. In some embodiments, administration of the compounds disclosed herein with food enhances one or more pharmacokinetic properties compared to administration under fasting conditions.

[0110] In the methods disclosed herein, administration of the disclosed compound with food enhances one or more pharmacokinetic properties of the compound. In the methods disclosed herein, administration of the disclosed compound with food enhances one or more pharmacokinetic properties of the compound compared to administration under fasting conditions.

[0111] In some embodiments, the enhanced pharmacokinetic properties are clinically significant. In some embodiments, the enhanced pharmacokinetic properties are increased pharmacokinetic properties. For example, in some embodiments, when administered with food, the C60 of the compounds disclosed herein... max Increase.

[0112] In some implementations, enhancing pharmacokinetic properties means increasing pharmacokinetic properties. For example, enhancing pharmacokinetic properties includes an increase in any one, two, three, or all of the following parameters: mean plasma concentration, C60, etc. max AUC, AUC (0-t) and / or AUC (inf) In some implementations, enhancing pharmacokinetic properties is equivalent to reducing pharmacokinetic properties.

[0113] In the methods disclosed herein, administration of the disclosed compounds with food increases the subject's exposure to the compounds. In the methods disclosed herein, administration of the disclosed compounds with food increases the exposure to the compounds compared to administration under fasting conditions. In some embodiments, the increased exposure is clinically significant.

[0114] The fasting subject can be, for example, a subject who fasts for a period of time prior to administration of a certain dose of the compound. Non-limiting examples of this time period include at least about 1 hour, at least about 2 hours, at least about 3 hours, at least about 4 hours, at least about 5 hours, at least about 6 hours, at least about 7 hours, at least about 8 hours, at least about 9 hours, at least about 10 hours, at least about 11 hours, at least about 12 hours, at least about 13 hours, at least about 14 hours, at least about 15 hours, or longer. In some embodiments, the subject is eligible to drink water or consume health products during fasting.

[0115] In some embodiments, the dosage is in a single dosage form or more than one dosage form (e.g., a 600 mg dose can be one 600 mg dosage form, two 300 mg dosage forms, three 200 mg dosage forms, six 100 mg dosage forms, etc.). In some embodiments, the dosage includes more than one pill administered simultaneously.

[0116] The subject may be someone who has consumed food for a period of time prior to drug administration. In some embodiments, the food is consumed within a certain timeframe before drug administration. In some embodiments, the subject has consumed food for a certain amount of time prior to administration of the compound of this disclosure. In some embodiments, the timeframe is approximately 5 minutes, approximately 10 minutes, approximately 15 minutes, approximately 20 minutes, approximately 25 minutes, approximately 30 minutes, approximately 35 minutes, approximately 40 minutes, approximately 45 minutes, approximately 50 minutes, approximately 55 minutes, approximately 1 hour, approximately 1.5 hours, or approximately 2 hours. In some embodiments, the compound disclosed herein is administered within approximately 0 to 60 minutes, approximately 0 to 30 minutes, or 5 to 20 minutes after food consumption. In some embodiments, the compound disclosed herein is administered 60 minutes, 30 minutes, 25 minutes, 20 minutes, 15 minutes, 10 minutes, or 5 minutes after food consumption.

[0117] In some embodiments, one or more pharmacokinetic properties are greater in subjects who have eaten compared to those in fasting subjects. In some embodiments, the mean plasma concentration is increased in subjects who have eaten compared to those in fasting subjects. In some embodiments, the Cg of subjects who have eaten is greater than that of subjects who have fasted. max Compared to the subjects who ate, C max Increase. In some implementations, the T of fasting subjects is increased. max Compared to the T of the food-eating subjects max Increase. In some implementations, the AUC of fasting subjects is increased. 0-last Compared to the AUC of the food-eating subjects 0-last Increase. In some implementations, the AUC of fasting subjects is increased. 0-Inf Compared to the AUC of the food-eating subjects 0-Inf Increase.

[0118] In some implementations, one or more pharmacokinetic properties of subjects who have eaten are increased by approximately 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, or more compared to one or more pharmacokinetic properties of subjects who have fasted.

[0119] In some implementations, one or more pharmacokinetic properties are greater in fasting subjects compared to those in subjects that have eaten. In some implementations, the plasma CL / F is greater in fasting subjects compared to those in subjects that have eaten. In some implementations, the Vc is greater in fasting subjects compared to those in subjects that have eaten. z Compared to F, the V of the fasting subjects z / F is larger.

[0120] In the methods disclosed herein, one or more pharmacokinetic properties of the compound in fasting subjects are not altered compared to those in subjects who have eaten. In some embodiments, the total urinary excretion of the compound in fasting subjects is not altered over a 96-hour interval compared to the total urinary excretion of the compound in subjects who have eaten. In some embodiments, the total urinary excretion of the compound in fasting subjects is not altered compared to that in subjects who have eaten. R Compared to fasting subjects, CL R It remained unchanged over a 96-hour interval.

[0121] In some implementations, the participants are of Caucasian ethnicity. In some implementations, the participants are of non-Asian ethnicity. In some implementations, the participants are of Asian ethnicity. In some implementations, the participants are of Japanese ethnicity.

[0122] In some embodiments, subjects of Japanese ethnicity exhibit one or more pharmacokinetic properties that are greater than those of subjects of Caucasian or non-Asian ethnicity. In some embodiments, subjects of Japanese ethnicity exhibit greater mean plasma concentrations than subjects of Caucasian or non-Asian ethnicity. In some embodiments, C0.05 is greater than that of subjects of Caucasian or non-Asian ethnicity. max In comparison, the C of subjects of the Japanese race max Larger. In some implementations, T values ​​were compared with those of subjects of Caucasian or non-Asian ethnicities. max In comparison, the T-scores of subjects of the Japanese race were... max Larger. In some implementations, the AUC is compared with that of subjects of Caucasian or non-Asian ethnicities. 0-last In comparison, the AUC of subjects of the Japanese race 0-last Larger. In some implementations, the AUC is compared with that of subjects of Caucasian or non-Asian ethnicities. 0-Inf In comparison, the AUC of subjects of the Japanese race 0-Inf Larger.

[0123] In some implementations, the Japanese participant is a participant whose parents and grandparents are both Japanese and were born in Japan. In some implementations, the Japanese participant is a participant whose parents and grandparents are both Japanese. In some implementations, the Japanese participant is a participant whose parents and grandparents are both Japanese.

[0124] In some embodiments, one or more pharmacokinetic properties are greater in subjects who are fasting compared to those in subjects who are fasting or of non-Asian ethnicity. In some embodiments, the mean plasma concentrations are greater in subjects who are fasting or of non-Asian ethnicity. In some embodiments, the C60 concentrations are greater in subjects who are fasting or of non-Asian ethnicity. max In comparison, the C-level results of food intake subjects who were Caucasian or of non-Asian races were significantly lower. max Larger. In some implementations, the AUC of fasting subjects in Caucasian or non-Asian races is larger than that of fasting subjects. In some implementations, the T... max In comparison, the T values ​​of food-eating subjects who were Caucasian or of non-Asian races were significantly lower. max extend.

[0125] Non-limiting examples of treatment-related adverse events (TRAEs) that can be used to determine the effectiveness of treating a subject with the compositions of this disclosure include: nausea, vomiting, increased serum creatinine, diarrhea, fatigue, increased ALT, increased AST, anemia, decreased appetite, proteinuria, rash maculo-papular rash, headache, increased lipase, decreased platelet count, increased amylase, and dehydration.

[0126] In some implementations, TRAEs are self-reported by the subjects. In other implementations, TRAEs are observed by health practitioners.

[0127] In some embodiments, the TRAE is a gastrointestinal (GI) TRAE. Non-limiting examples of GI TRAEs that can be used to determine the effect of treating a subject with the compositions of this disclosure include nausea, vomiting, and diarrhea. In some embodiments, the GI TRAE is nausea. In some embodiments, the GI TRAE is vomiting. In some embodiments, the GI TRAE is diarrhea.

[0128] In some implementations, gastrointestinal adverse events are self-reported by the subjects. In other implementations, gastrointestinal adverse events are observed by a healthcare professional.

[0129] In some embodiments, the frequency of one or more TRAE rates is lower after administration of the compounds of this disclosure to feeding subjects compared to the rates of one or more TRAEs in fasting subjects. In some embodiments, the frequency of GI TRAE rates is lower after administration of the compounds of this disclosure to feeding subjects compared to the GI TRAE rates in fasting subjects.

[0130] In some embodiments, the compounds disclosed herein, when administered with food, result in an improvement in one or more adverse gastrointestinal events (TRAEs), such as nausea, vomiting, or diarrhea.

[0131] In some implementations, the incidence of one or more GI TRAEs observed in feeding subjects is reduced compared to the incidence observed in fasting subjects. In some implementations, the incidence of one or more GI TRAEs observed in feeding subjects is reduced by approximately 15%, approximately 20%, approximately 25%, approximately 30%, approximately 35%, approximately 40%, approximately 45%, approximately 50%, approximately 55%, approximately 60%, approximately 65%, approximately 70%, approximately 75%, approximately 80%, approximately 85%, or more compared to the incidence observed in fasting subjects.

[0132] In some embodiments, the incidence of one or more GI TRAEs observed in feeding subjects is reduced by approximately 15% compared to the incidence observed in fasting subjects. In some embodiments, the incidence of one or more GI TRAEs observed in feeding subjects is reduced by approximately 20% compared to the incidence observed in fasting subjects. In some embodiments, the incidence of one or more GI TRAEs observed in feeding subjects is reduced by approximately 25% compared to the incidence observed in fasting subjects. In some embodiments, the incidence of one or more GI TRAEs observed in feeding subjects is reduced by approximately 30% compared to the incidence observed in fasting subjects. In some embodiments, the incidence of one or more GI TRAEs observed in feeding subjects is reduced by approximately 35% compared to the incidence observed in fasting subjects. In some embodiments, the incidence of one or more GI TRAEs observed in feeding subjects is reduced by approximately 40% compared to the incidence observed in fasting subjects. In some embodiments, the incidence of one or more GI TRAEs observed in feeding subjects is reduced by approximately 45% compared to the incidence observed in fasting subjects. In some embodiments, the incidence of one or more GI TRAEs observed in feeding subjects is reduced by approximately 50% compared to the incidence observed in fasting subjects. In some embodiments, the incidence of one or more GI TRAEs observed in feeding subjects is reduced by approximately 55% compared to the incidence observed in fasting subjects. In some embodiments, the incidence of one or more GI TRAEs observed in feeding subjects is reduced by approximately 60% compared to the incidence observed in fasting subjects. In some embodiments, the incidence of one or more GI TRAEs observed in feeding subjects is reduced by approximately 65% ​​compared to the incidence observed in fasting subjects.

[0133] In some embodiments, the food includes a high-fat diet. In some embodiments, the high-fat diet comprises at least 50% of the total calorie content of the diet as fat. In some embodiments, the high-fat diet comprises at least 500 kcal of fat. In some embodiments, the high-fat diet comprises at least 55 g of fat.

[0134] In some embodiments, the food comprises a moderate-fat diet. In some embodiments, a moderate-fat diet comprises approximately 25% to approximately 50% of the total calorie content of the diet as fat. In some embodiments, a moderate-fat diet comprises approximately 125 kcal to approximately 500 kcal of fat. In some embodiments, a moderate-fat diet comprises approximately 14 g to approximately 55 g of fat.

[0135] In some embodiments, the food includes a low-fat diet. In some embodiments, a low-fat diet comprises less than 25% of the total calorie content of the diet as fat. In some embodiments, a low-fat diet comprises less than about 125 kcal of fat from fat. In some embodiments, a low-fat diet comprises less than about 14 g of fat.

[0136] In some implementations, a high-fat diet contains approximately 150 calories from protein, approximately 250 calories from carbohydrates, and approximately 500 to 600 calories from fat.

[0137] In some embodiments, the food includes a high-calorie diet. In some embodiments, the high-calorie diet contains at least 800 calories. In some embodiments, the high-calorie diet contains approximately 800 calories to approximately 1000 calories.

[0138] In some embodiments, the food includes a high-fat, high-calorie diet. In some embodiments, a high-fat, high-calorie diet comprises at least 50% fat and approximately 800 to approximately 1000 calories of total calories.

[0139] In some implementations, if a therapeutically effective amount of the compound is administered to fasting study subjects and a therapeutically effective amount of the compound is administered to eating study subjects, then the C of the compound in the eating study subjects... max C of compounds greater than those in fasting study subjects max In some implementations, if a therapeutically effective amount of the compound is administered to a group of subjects who are eating and a group of subjects who are fasting, C0.05 levels of approximately 8350 ng / mL to approximately 12300 ng / mL are observed in the eating subjects. max Values, and C values ​​of 7200 ng / mL to approximately 11100 ng / mL were observed in fasting subjects. max Value. In some implementations, if a therapeutically effective amount of the compound is administered to a group of subjects who are eating and a group of subjects who are fasting, C values ​​of approximately 12,300 ng / mL to approximately 28,100 ng / mL are observed in the eating subjects. maxValues, and C values ​​of approximately 2820 ng / mL to 20100 ng / mL were observed in fasting subjects. max Value. In some implementations, if a therapeutically effective amount of the compound is administered to a group of fasting subjects, C values ​​of approximately 6710 ng / mL to approximately 20600 ng / mL are observed. max value.

[0140] In some implementations, if a therapeutically effective amount of the compound is administered to fasting study subjects and a therapeutically effective amount of the compound is administered to eating study subjects, then the T value of the compound in the eating study subjects is... max T of compounds greater than those in fasting study subjects max In some implementations, if a therapeutically effective amount of the compound is administered to a group of eating study subjects and a group of fasting subjects, a T value of approximately 1.51 h to approximately 5.01 h is observed in the eating subjects. max The value was observed, and T values ​​were observed from 1.54 h to approximately 2.51 h in fasting subjects. max Value. In some implementations, if a therapeutically effective amount of the compound is administered to a group of feeding study subjects and a group of fasting subjects, a T value of approximately 2.11 h to approximately 8.08 h is observed in the feeding subjects. max The value was observed, and T values ​​were observed from 1.03 h to approximately 5.94 h in fasting subjects. max Values. In some implementations, if a therapeutically effective amount of the compound is administered to a group of fasting subjects, T values ​​are observed from approximately 1.59 h to approximately 5.06 h. max value.

[0141] In some implementations, if a therapeutically effective amount of the compound is administered to fasting study subjects and a therapeutically effective amount of the compound is administered to eating study subjects, then the t-value of the compound in the eating study subjects is... 1 / 2 The t of compounds greater than those in fasting study subjects 1 / 2 In some implementations, if a therapeutically effective amount of the compound is administered to a group of subjects who are eating and a group of subjects who are fasting, a t-reduction period of approximately 11.9 h to approximately 21.7 h is observed in the eating subjects. 1 / 2 The value was observed, and in fasting subjects, the t-value ranged from approximately 13.6 h to 20.8 h. 1 / 2 Value. In some implementations, if a therapeutically effective amount of the compound is administered to a group of feeding study subjects and a group of fasting subjects, a t-value of approximately 13.5 h to approximately 38.2 h is observed in the feeding subjects. 1 / 2 The value was observed in fasting subjects from 11.5 h to approximately 45.8 h. 1 / 2Value. In some implementations, if a therapeutically effective amount of the compound is administered to a group of fasting subjects, a t-value is observed from approximately 11.2 h to approximately 30.0 h. 1 / 2 value.

[0142] In some implementations, if a therapeutically effective amount of the compound is administered to fasting study subjects and a therapeutically effective amount of the compound is administered to eating study subjects, the AUC of the compound in the eating study subjects is greater than the AUC of the compound in the fasting study subjects.

[0143] In some implementations, AUC is AUC 0-inf In some implementations, if a therapeutically effective amount of the compound is administered to a group of subjects who are eating and a group of subjects who are fasting, an AUC of approximately 114,000 h·ng / mL to approximately 264,000 h·ng / mL is observed in the eating subjects. 0-inf Values, and AUCs of approximately 98,400 h·ng / mL to approximately 183,000 h·ng / mL were observed in fasting subjects. 0-inf Values. In some implementations, if a therapeutically effective amount of the compound is administered to a group of feeding study subjects and a group of fasting subjects, an AUC of approximately 272,000 h·ng / mL to approximately 839,000 h·ng / mL is observed in the feeding subjects. 0-inf Values, and AUCs of 106,000 h·ng / mL to approximately 613,000 h·ng / mL were observed in fasting subjects. 0-inf Values. In some implementations, if a therapeutically effective amount of the compound is administered to a group of fasting subjects, an AUC of approximately 209,000 h·ng / mL to approximately 487,000 h·ng / mL is observed. 0-inf value.

[0144] In some implementations, AUC is AUC 0-last In some implementations, if a therapeutically effective amount of the compound is administered to a group of subjects who are eating and a group of subjects who are fasting, an AUC of approximately 113,000 h·ng / mL to approximately 253,000 h·ng / mL is observed in the eating subjects. 0-last Values ​​were observed, and AUCs of approximately 95,900 h·ng / mL to 182,000 h·ng / mL were observed in fasting subjects. 0-last Values. In some implementations, if a therapeutically effective amount of the compound is administered to a group of feeding study subjects and a group of fasting subjects, an AUC of approximately 269,000 h·ng / mL to approximately 697,000 h·ng / mL is observed in the feeding subjects. 0-lastValues, and AUCs of approximately 105,000 h·ng / mL to approximately 555,000 h·ng / mL were observed in fasting subjects. 0-last Values. In some implementations, if a therapeutically effective amount of the compound is administered to a group of fasting subjects, an AUC of approximately 201,000 h·ng / mL to approximately 481,000 h·ng / mL is observed. 0-last value.

[0145] In some implementations, AUC is AUC 0-24 In some implementations, if a therapeutically effective amount of the compound is administered to a group of feeding study subjects and a group of fasting subjects, an AUC of approximately 85,800 h·ng / mL to approximately 152,000 h·ng / mL is observed in the feeding subjects. 0-24 The values ​​were observed to be approximately 72,900 h·ng / mL to 121,000 h·ng / mL in fasting subjects. 0-24 Values. In some implementations, if a therapeutically effective amount of the compound is administered to a group of subjects who are eating and a group of subjects who are fasting, an AUC of approximately 178,000 h·ng / mL to approximately 347,000 h·ng / mL is observed in the eating subjects. 0-24 Values, and AUCs of approximately 46,900 h·ng / mL to approximately 277,000 h·ng / mL were observed in fasting subjects. 0-24 Values. In some implementations, if a therapeutically effective amount of the compound is administered to a group of fasting subjects, an AUC of approximately 97,300 h·ng / mL to approximately 283,000 h·ng / mL is observed. 0-24 value.

[0146] In some implementations, AUC is AUC 0-96 In some implementations, if a therapeutically effective amount of the compound is administered to a group of subjects who are eating and a group of subjects who are fasting, an AUC of approximately 114,000 h·ng / mL to approximately 253,000 h·ng / mL is observed in the eating subjects. 0-96 Values ​​were observed, and AUCs of approximately 97,600 h·ng / mL to approximately 182,000 h·ng / mL were observed in fasting subjects. 0-96 value.

[0147] In some implementations, AUC is AUC %extrap In some implementations, if a therapeutically effective amount of the compound is administered to a group of subjects who are eating and a group of subjects who are fasting, an AUC of approximately 0.884% to approximately 4.21% is observed in the eating subjects. %extrapThe values ​​were observed in fasting subjects at approximately 0.821% to 3.62% of the AUC. %extrap Value. In some implementations, if a therapeutically effective amount of the compound is administered to a group of eating study subjects and a group of fasting subjects, an AUC of approximately 0.953% to approximately 16.8% is observed in the eating subjects. %extrap The values ​​were observed in fasting subjects, with AUCs ranging from 0.901% to approximately 9.51%. %extrap Values. In some implementations, if a therapeutically effective amount of the compound is administered to a group of fasting subjects, an AUC of approximately 0.482% to approximately 10.5% is observed. %extrap value.

[0148] In some embodiments, if a therapeutically effective amount of the compound is administered to both fasting and feeding subjects, the CL / F of the compound is greater in the fasting subjects than in the feeding subjects. In some embodiments, if a therapeutically effective amount of the compound is administered to both a feeding group and a fasting group, a CL / F value of approximately 3.79 L / h to approximately 8.76 L / h is observed in the feeding subjects, and a CL / F value of approximately 5.45 L / h to approximately 10.2 L / h is observed in the fasting subjects. In some embodiments, if a therapeutically effective amount of the compound is administered to both a feeding group and a fasting group, a CL / F value of approximately 2.39 L / h to approximately 7.36 L / h is observed in the feeding subjects, and a CL / F value of approximately 3.27 L / h to approximately 18.8 L / h is observed in the fasting subjects. In some implementations, if a therapeutically effective amount of the compound is administered to a group of fasting subjects, CL / F values ​​of approximately 4.11 L / h to approximately 9.56 L / h are observed.

[0149] In some implementations, if a therapeutically effective amount of the compound is administered to fasting study subjects and a therapeutically effective amount of the compound is administered to eating study subjects, then the V of the compound in the fasting study subjects... Z / F is greater than the V of compounds in subjects of the food intake study. Z / F. In some implementations, if a therapeutically effective amount of the compound is administered to a group of feeding study subjects and a group of fasting subjects, a volume of approximately 99.6 L to approximately 164 L is observed in the feeding subjects. Z / F value, and V values ​​of 107 L to approximately 199 L were observed in fasting subjects. Z / F value. In some implementations, if a therapeutically effective amount of the compound is administered to a group of feeding study subjects and a group of fasting subjects, a VF of approximately 86.8 L to approximately 216 L is observed in the feeding subjects.Z / F value, and V values ​​of 117 L to approximately 382 L were observed in fasting subjects. Z / F value. In some implementations, if a therapeutically effective amount of the compound is administered to a group of fasting subjects, a VF of approximately 93.8 L / h to approximately 251 L / h is observed. Z / F value.

[0150] In some implementations, if a therapeutically effective amount of the compound is administered to fasting study subjects and a therapeutically effective amount of the compound is administered to eating study subjects, the mean peak plasma concentration (C0) of the compound is... max The ratio of the compound's concentration to its mean 12-hour plasma concentration (C12h) was greater than the compound's mean peak plasma concentration (C12h) under fasting conditions. max The ratio of plasma concentration to mean 12-hour plasma concentration (C12h).

[0151] Pharmaceutically acceptable salts This disclosure provides for the use of pharmaceutically acceptable salts of any of the therapeutic compounds described herein. Pharmaceutically acceptable salts include, for example, acid addition salts and base addition salts. The acid added to the compound to form an acid addition salt can be an organic or inorganic acid. The base added to the compound to form a base addition salt can be an organic or inorganic base. In some embodiments, the pharmaceutically acceptable salt is a metal salt. In some embodiments, the pharmaceutically acceptable salt is an ammonium salt.

[0152] Metal salts can be produced by adding an inorganic base to the compounds of this disclosure. The inorganic base consists of a metal cation paired with a basic counterion, such as, for example, hydroxide, carbonate, bicarbonate, or phosphate. The metal can be an alkali metal, alkaline earth metal, transition metal, or main group metal. In some embodiments, the metal is lithium, sodium, potassium, cesium, cerium, magnesium, manganese, iron, calcium, strontium, cobalt, titanium, aluminum, copper, cadmium, or zinc.

[0153] In some implementations, the metal salt is a lithium salt, sodium salt, potassium salt, cesium salt, cerium salt, magnesium salt, manganese salt, iron salt, calcium salt, strontium salt, cobalt salt, titanium salt, aluminum salt, copper salt, cadmium salt, or zinc salt.

[0154] Ammonium salts can be produced by adding ammonia or an organic amine to the compounds disclosed herein. In some embodiments, the organic amine is triethylamine, diisopropylamine, ethanolamine, diethanolamine, triethanolamine, morpholine, N-methylmorpholine, piperidine, N-methylpiperidine, N-ethylpiperidine, dibenzylamine, piperazine, pyridine, pyrazole, imidazole, or pyrazine.

[0155] In some embodiments, the ammonium salt is a triethylamine salt, trimethylamine salt, diisopropylamine salt, ethanolamine salt, diethanolamine salt, triethanolamine salt, morpholine salt, N-methylmorpholine salt, piperidine salt, N-methylpiperidine salt, N-ethylpiperidine salt, dibenzylamine salt, piperazine salt, pyridine salt, pyrazole salt, pyridazine salt, pyrimidine salt, imidazole salt, or pyrazine salt.

[0156] Acid addition salts can be generated by adding an acid to a compound of the present disclosure. In some embodiments, the acid is organic. In some embodiments, the acid is inorganic. In some embodiments, the acid is hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, nitrous acid, sulfuric acid, sulfurous acid, phosphoric acid, isonicotinic acid, lactic acid, salicylic acid, tartaric acid, ascorbic acid, gentian acid, gluconic acid, glucuronic acid, saccharic acid, formic acid, benzoic acid, glutamic acid, pantothenic acid, acetic acid, propionic acid, butyric acid, fumaric acid, succinic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, oxalic acid, or maleic acid.

[0157] In some embodiments, the salt is a hydrochloride, hydrobromide, hydroiodide, nitrate, nitrite, sulfate, sulfite, phosphate, isonicotinate, lactate, salicylate, tartrate, ascorbate, gentianate, gluconate, glucuronate, glucodinolate, formate, benzoate, glutamate, pantothenate, acetate, propionate, butyrate, fumarate, succinate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, citrate, oxalate, or maleate.

[0158] The pharmaceutical compositions disclosed herein The pharmaceutical compositions disclosed herein may be used, for example, before, during, or after treatment of a subject with another pharmaceutical agent.

[0159] Subjects may be, for example, elderly people, adults, adolescents, pre-adolescent children, children, toddlers, infants, newborns, and non-human animals. In some implementations, subjects are patients.

[0160] The pharmaceutical compositions disclosed herein can be combinations of any pharmaceutical compound described herein with other chemical components such as carriers, stabilizers, diluents, dispersants, suspending agents, thickeners, and / or excipients. The pharmaceutical compositions facilitate the administration of the compound to a living organism. The pharmaceutical compositions can be administered in therapeutically effective amounts via various forms and routes, including, for example, intravenous administration, subcutaneous administration, intramuscular administration, oral administration, parenteral administration, ocular administration, subcutaneous administration, transdermal administration, nasal administration, vaginal administration, and topical administration.

[0161] The pharmaceutical composition can be administered topically, for example, by direct injection of the compound into an organ, optionally as a reservoir formulation, a sustained-release formulation, or an implant. The pharmaceutical composition can be provided as a rapid-release formulation, a prolonged-release formulation, or an intermediate-release formulation. Rapid-release formulations provide immediate release. Prolonged-release formulations provide controlled release or sustained, delayed release.

[0162] For oral administration, pharmaceutical compositions can be formulated by combining the active compound with a pharmaceutically acceptable carrier or excipient. Such carriers can be used to formulate liquids, gels, syrups, elixirs, slurries, or suspensions for oral ingestion by a subject. Non-limiting examples of solvents for oral soluble formulations include water, ethanol, isopropanol, saline, physiological saline, DMSO, dimethylformamide, potassium phosphate buffer, phosphate-buffered saline (PBS), sodium phosphate buffer, 4-2-hydroxyethyl-1-piperazine ethanesulfonic acid buffer (HEPES), 3-(N-morpholino)propanesulfonic acid buffer (MOPS), piperazine-N,N'-bis(2-ethanesulfonic acid) buffer (PIPES), and saline-sodium citrate buffer (SSC). Non-limiting examples of co-solvents used in oral soluble formulations include sucrose, urea, cremaphore, DMSO, and potassium phosphate buffer.

[0163] Pharmaceutical preparations can be formulated for intravenous administration. Pharmaceutical compositions can be in the form of sterile suspensions, solutions, or emulsions suitable for use as oily or aqueous media for parenteral injection, and may contain a formulation agent, such as a suspending agent, stabilizer, and / or dispersant. Pharmaceutical preparations for parenteral administration include aqueous solutions of the active compound in a water-soluble form. Suspensions of the active compound can be prepared as oily injectable suspensions. Suitable lipophilic solvents or media include fatty oils such as sesame oil, or synthetic fatty acid esters such as ethyl oleate or triglycerides, or liposomes. Suspensions may also contain suitable stabilizers or agents that increase the solubility of the compound to allow for the preparation of highly concentrated solutions. Alternatively, the active ingredient can be in powder form for preparation with a suitable media, such as sterile pyrogen-free water, prior to use.

[0164] The active compound can be applied topically and can be formulated into a variety of topical compositions, such as solutions, suspensions, lotions, gels, pastes, medicated sticks, balms, creams, and ointments. Such pharmaceutical compositions may contain solubilizers, stabilizers, tension enhancers, buffers, and preservatives.

[0165] The compounds of this disclosure can be applied topically to the skin or body cavity of a subject, such as the oral cavity, vaginal cavity, bladder cavity, cranial cavity, spinal cavity, thoracic cavity, or pelvic cavity. The compounds of this disclosure can be applied to accessible body cavities.

[0166] The compounds can also be formulated into rectal compositions, such as enemas, rectal gels, rectal foams, rectal aerosols, suppositories, gel suppositories, or retention enemas, comprising a conventional suppository base such as cocoa butter or other glycerides, and synthetic polymers such as polyvinylpyrrolidone and PEG. In the suppository form of the composition, a low-melting-point wax, such as a mixture of fatty acid glycerides optionally combined with cocoa butter, can be melted.

[0167] When practicing the treatments or methods of use provided herein, a therapeutically effective amount of the compounds described herein is administered as a pharmaceutical composition to a subject suffering from the disease or condition to be treated. In some embodiments, the subject is a mammal, such as a human. The therapeutically effective amount can vary widely depending on the severity of the disease, the age and relative health of the subject, the potency of the compound used, and other factors. The compounds can be used alone or in combination with one or more therapeutic agents as components of a mixture.

[0168] Pharmaceutical compositions can be formulated using one or more physiologically acceptable carriers comprising excipients and adjuvants, which facilitate the processing of the active compound into a pharmaceutically usable article. The formulation can be modified depending on the chosen route of administration. Pharmaceutical compositions containing the compounds described herein can be manufactured, for example, by mixing, dissolving, emulsifying, encapsulating, embedding, or compressing processes.

[0169] Pharmaceutical compositions may comprise at least one pharmaceutically acceptable carrier, diluent, or excipient, and the compounds described herein as either a free base or a pharmaceutically acceptable salt. Such pharmaceutical compositions may also comprise solubilizers, stabilizers, tension enhancers, buffers, and preservatives.

[0170] Methods for preparing compositions comprising the compounds described herein include formulating the compounds with one or more inert, pharmaceutically acceptable excipients or carriers to form solid compositions, semi-solid compositions, or liquid compositions. Solid compositions include, for example, powders, tablets, dispersible granules, capsules, and granules. Liquid compositions include, for example, solutions in which the compounds are dissolved, emulsions containing the compounds, or solutions containing liposomes, micelles, or nanoparticles containing the compounds disclosed herein. Semi-solid compositions include, for example, gels, suspensions, and creams. Compositions may be in the form of liquid solutions or suspensions, suitable for solid forms dissolved or suspended in a liquid prior to use, or as emulsions. These compositions may also contain small amounts of non-toxic excipients, such as wetting agents or emulsifiers, pH buffers, and other pharmaceutically acceptable additives.

[0171] Non-limiting examples of dosage forms suitable for use in this disclosure include liquids, powders, gels, nanosuspensions, nanoparticles, microgels, aqueous or oily suspensions, emulsions, and any combination thereof.

[0172] Non-limiting examples of pharmaceutically acceptable excipients suitable for use in this disclosure include binders, disintegrants, anti-adhesion agents, antistatic agents, surfactants, antioxidants, coating agents, colorants, plasticizers, preservatives, suspending agents, emulsifiers, antimicrobial agents, spheronization agents, and any combination thereof.

[0173] The compositions of this disclosure can be, for example, immediate-release forms or controlled-release formulations. Immediate-release formulations can be formulated to allow the compound to act rapidly. Non-limiting examples of immediate-release formulations include readily soluble formulations. Controlled-release formulations can be pharmaceutical formulations that have been adapted such that the release rate and profile of the active agent can be matched to physiological and time-dependent therapeutic requirements, or optionally have been formulated to achieve a programmed release of the active agent. Non-limiting examples of controlled-release formulations include granules, delayed-release granules, hydrogels (e.g., hydrogels of synthetic or natural origin), other gelling agents (e.g., gel-forming dietary fibers), matrix-based formulations (e.g., formulations comprising a polymeric material having at least one active ingredient dispersed therein), matrix-integrated granules, polymer mixtures, and granular masses.

[0174] In some embodiments, the controlled-release formulation is a delayed-release form. The delayed-release form can be formulated to prolong the duration of the compound's effect. The delayed-release form can be formulated to delay the release of one or more compounds of an effective dose, for example, for about 4 hours, about 8 hours, about 12 hours, about 16 hours, or about 24 hours.

[0175] Controlled-release formulations can be in a sustained-release form. Sustained-release forms can be formulated to provide, for example, the action of a compound over an extended period of time. Sustained-release forms can be formulated to provide an effective dose of any compound described herein (e.g., to provide a physiologically effective blood profile) over about 4 hours, about 8 hours, about 12 hours, about 16 hours, or about 24 hours.

[0176] Non-restrictive examples of pharmaceutically acceptable excipients can be found, for example, in Remington: The Science and Practice of Pharmacy, 19th edition (Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania 1975; Liberman, HA and Lachman, L. (eds.), Pharmaceutical Dosage Forms, Marcel Decker, New York, NY, 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, 7th edition (Lippincott Williams & Wilkins 1999), each of which is incorporated in its entirety by reference.

[0177] Multiple therapeutic agents can be administered in any order or simultaneously. In some embodiments, the compounds of this disclosure are administered in combination with another therapeutic agent, before or after treatment with the other therapeutic agent. If administered simultaneously, the multiple therapeutic agents can be provided in a single, uniform form or in multiple forms, such as as multiple separate pills. The agents can be packaged together or separately in a single package or in more than one package. One or all of the therapeutic agents can be administered in multiple doses. If not administered simultaneously, the timing between the multiple doses can vary for up to about one month.

[0178] In some embodiments, the compounds of this disclosure are administered in combination with an antiemetic, either before or after treatment with the antiemetic. Suitable antiemetics include, but are not limited to: 5-HT3 receptor antagonists, such as ondansetron, granisetron, dolasetron, and palonosetron; NK1 receptor antagonists, such as aprepitant and fosaprepitant; corticosteroids, such as dexamethasone; dopamine receptor antagonists, such as phenothiazines (e.g., prochlorperazine), butyrophenones (e.g., haloperidol, fluperidone), and substituted benzamides (e.g., metoclopramide); benzodiazepines, such as lorazepam and alprazolam; olanzapine; and cannabinoids, such as dronabinol and cannabinol.

[0179] In some embodiments, the compounds of this disclosure are administered in combination with a gastric acid-reducing agent, either before or after treatment with the gastric acid-reducing agent. In some embodiments, the gastric acid-reducing agent is an H2-receptor antagonist. In some embodiments, the H2-receptor antagonist is cimetidine, ranitidine, famotidine, nizatidine, roxatidine, lafutidine, lavotidine, niperotidine, or sufotidine. In some embodiments, the H2-receptor antagonist is famotidine. In some embodiments, the gastric acid-reducing agent is a proton pump inhibitor. In some embodiments, the proton pump inhibitor is omeprazole, esomeprazole, lansoprazole, dexlansoprazole, pantoprazole, or rabeprazole. In some implementations, the proton pump inhibitor is rabeprazole.

[0180] In some embodiments, this disclosure provides a method for treating a subject in need of treatment, the method comprising selecting subjects for treatment based on the fact that the subject has not been treated with a proton pump inhibitor within 7 days prior to administering the p53 activating compound of this disclosure, and administering the p53 activating compound to the subject based on said selection. In some embodiments, the method includes administering an H2-receptor antagonist to the subject within 7 days prior to administering the p53 activating compound.

[0181] In some embodiments, this disclosure provides a method for treating a subject in need of treatment, the method comprising: a) selecting a subject to be treated with a p53-activating compound of this disclosure based on the subject not having been treated with a CYP3A4 modulator, P-gp inhibitor, or BCRP inhibitor, and b) administering the p53-activating compound to the subject based on said selection. CYP3A4 modulators include CYP3A4 inhibitors such as, for example, clarithromycin, cobicistat, elvitegravir and ritonavir, grapefruit juice, indinavir and ritonavir, ederaris, itraconazole, ketoconazole, lopinavir and ritonavir, nefazodone, nelfinavir, paritaprevir / ritonavir / ombitasvir, and / or dasabuvir. Asabuvir, posaconazole, ritonavir, saquinavir and ritonavir, telazalovir, telithromycin and voriconazole; and CYP3A4 inducers, such as apalutamide, carbamazepine, enzalutamide, mitotan, phenytoin, rifampin and St. John's wort. P-gp inhibitors include, for example, amiodarone, carvedilol, clarithromycin, dronedarone, itraconazole, lopinavir and ritonavir, propafenone, quinidine, ranolazine, ritonavir, saquinavir and ritonavir, telazalovir and ritonavir, and verapamil. BCRP inhibitors include, for example, curcumin, cyclosporine A, and eltrombopag.

[0182] In some embodiments, this disclosure provides a method for treating a subject with a corresponding need, the method comprising switching the subject from treatment with a proton pump inhibitor to treatment with an H2-receptor antagonist, and administering the p53 activating compound of this disclosure to the subject no earlier than 7 days after the switch.

[0183] The therapeutic agents described herein can be administered before, during, or after the onset of a disease or condition, and the timing of administration of the composition containing the therapeutic agent can vary. For example, the composition can be used as a prophylactic agent and can be administered continuously to subjects predisposed to a condition or disease in order to reduce the likelihood of the disease or condition occurring. The composition can be administered to the subject as soon as possible during or after the onset of symptoms. Administration of the therapeutic agent can be initiated within 48 hours before the onset of symptoms, within 24 hours before the onset of symptoms, within 6 hours before the onset of symptoms, or within 3 hours of the onset of symptoms. Initial administration can be performed using any of the formulations described herein via any feasible route, such as those described herein.

[0184] The compound can be administered as soon as practicable after the onset of a disease or condition is detected or suspected, and for the duration necessary to continue treatment, such as, for example, from about 1 month to about 3 months. In some embodiments, the duration for which the compound can be administered can be about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 1 month, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 2 months, about 9 weeks, about 10 weeks, about 11 weeks, about 12 weeks, about 3 months, about 13 weeks, about 14 weeks, about 15 weeks, about 16 weeks, about 4 months, about 17 weeks, about 18 weeks, about 19 weeks, about 20 weeks, about 5 months, about 21 weeks, and so on. Treatment durations are estimated as follows: 22 weeks, approximately 23 weeks, approximately 24 weeks, approximately 6 months, approximately 7 months, approximately 8 months, approximately 9 months, approximately 10 months, approximately 11 months, approximately 1 year, approximately 13 months, approximately 14 months, approximately 15 months, approximately 16 months, approximately 17 months, approximately 18 months, approximately 19 months, approximately 20 months, approximately 21 months, approximately 22 months, approximately 23 months, approximately 2 years, approximately 2.5 years, approximately 3 years, approximately 3.5 years, approximately 4 years, approximately 4.5 years, approximately 5 years, approximately 6 years, approximately 7 years, approximately 8 years, approximately 9 years, or approximately 10 years. The duration of treatment may vary for each individual subject.

[0185] The pharmaceutical compositions described herein can be presented in unit dosage forms suitable for precise single-dose administration. In a unit dosage form, the formulation is divided into unit doses containing appropriate amounts of one or more compounds. The unit doses can be in the form of packages containing discrete amounts of the formulation. Non-limiting examples are packages of injectable, vials, or ampoules. Aqueous suspension compositions can be packaged in single-dose, non-reclosable containers. Multi-dose, reclosable containers can be used, for example, with or without preservatives. Formulations for injection can be presented in unit dosage forms, for example, in ampoules or in multi-dose containers with preservatives.

[0186] The pharmaceutical compositions described herein can be administered in combination with other therapies, such as chemotherapy, radiation, surgery, anti-inflammatory agents, and selected vitamins. Other agents may be administered before, after, or concurrently with the pharmaceutical compositions.

[0187] Depending on the intended mode of administration, the pharmaceutical composition may be in the form of a solid, semi-solid, or liquid dosage form, such as, for example, tablets, suppositories, pills, capsules, powders, liquids, suspensions, lotions, creams, or gels, for example, in a unit dosage form suitable for a precise dose for a single administration.

[0188] For solid compositions, non-toxic solid carriers include, for example, pharmaceutical-grade mannitol, lactose, starch, magnesium stearate, sodium saccharin, talc, cellulose, glucose, sucrose, and magnesium carbonate.

[0189] Non-limiting examples of pharmaceutically active agents suitable for use in combination with compositions of this disclosure include anti-infective agents (i.e., aminoglycosides), antiviral agents, antimicrobial agents, anticholinergic / antispasmodic agents, antidiabetic agents, antihypertensive agents, antitumor agents, cardiovascular agents, central nervous system agents, coagulation modulators, hormones, immunomodulators, immunosuppressants, and ophthalmic products.

[0190] Compounds can be delivered via liposome technology. Using liposomes as drug carriers can increase the therapeutic index of compounds. Liposomes contain natural phospholipids and may contain mixed lipid chains with surfactant properties (e.g., lecithin ethanolamine). Liposomes can be designed with surface ligands for attachment to unhealthy tissues. Non-limiting examples of liposomes include multilayered vesicles (MLVs), small unlayered vesicles (SUVs), and large unlayered vesicles (LUVs). The physicochemical properties of liposomes can be tuned to optimize penetration across biological barriers and retention at the application site, and to reduce the likelihood of premature degradation and toxicity to non-target tissues. Optimal liposome properties depend on the route of administration: larger liposomes show good retention upon local injection, while smaller liposomes are better suited for passive targeting. Polyethylene glycolization reduces hepatic and splenic uptake of liposomes and increases circulation time, resulting in increased localization at sites of inflammation due to enhanced permeability and retention (EPR) effects. Additionally, the liposome surface can be modified to selectively deliver encapsulated drugs to specific target cells. Non-limiting examples of targeted ligands include monoclonal antibodies, vitamins, peptides, and polysaccharides that are specific to receptors concentrated on the cell surface associated with disease.

[0191] Non-limiting examples of dosage forms suitable for use in this disclosure include liquids, elixirs, nanosuspensions, aqueous or oily suspensions, drops, syrups, and any combination thereof. Non-limiting examples of pharmaceutically acceptable excipients suitable for use in this disclosure include granulators, binders, lubricants, disintegrants, sweeteners, gliding agents, anti-adhesion agents, antistatic agents, surfactants, antioxidants, gums, coating agents, colorants, flavoring agents, plasticizers, preservatives, suspending agents, emulsifiers, plant cellulose materials, and pelleting agents, and any combination thereof.

[0192] The compositions disclosed herein can be packaged as kits. In some embodiments, the kit includes written instructions for the administration / use of the composition. The written material may be, for example, a label. The written material may suggest the conditions and methods of administration. The instructions provide best guidance to the subject and supervising physician for achieving optimal clinical outcomes from the administration of the therapy. The written material may be a label. In some embodiments, the label may be approved by a regulatory agency, such as the U.S. Food and Drug Administration (FDA), the European Medicines Agency (EMA), or other regulatory bodies.

[0193] Dosage The pharmaceutical compositions described herein can be presented in unit dosage forms suitable for precise single-dose administration. In a unit dosage form, the formulation is divided into unit doses containing appropriate amounts of one or more compounds. The unit doses can be in the form of packages containing discrete amounts of the formulation. Non-limiting examples are liquids in vials or ampoules. Aqueous suspension compositions can be packaged in single-dose non-reclosable containers. Multi-dose reclosable containers can be used, for example, in combination with preservatives. Formulations for parenteral injection can be presented in unit dosage forms, for example, in ampoules or in multi-dose containers with preservatives.

[0194] The compounds described herein may be available in quantities ranging from about 1 mg to about 3000 mg, from about 10 mg to about 3000 mg, from about 100 mg to about 3000 mg, from about 1000 mg to about 3000 mg, from 1 mg to about 2000 mg, from about 10 mg to about 2000 mg, from about 100 mg to about 2000 mg, from about 100 mg to about 2000 mg, from about 5 mg to about 1000 mg, from about 10 mg to about 500 mg, from about 50 mg to about 250 mg, from about 100 mg to about 200 mg, from about 1 mg to about 50 mg, from about 50 mg to about 100 mg, from about 100 mg to about 150 mg, from about 150 mg to about 200 mg, from about 200 mg to about 250 mg, from about 250 mg to about 300 mg, from about 300 mg to about 350 mg, from about 350 mg to about 400 mg, from about 400 mg to about 300 mg. The composition is present in the range of about 450 mg, from about 450 mg to about 500 mg, from about 500 mg to about 550 mg, from about 550 mg to about 600 mg, from about 600 mg to about 650 mg, from about 650 mg to about 700 mg, from about 700 mg to about 750 mg, from about 750 mg to about 800 mg, from about 800 mg to about 850 mg, from about 850 mg to about 900 mg, from about 900 mg to about 950 mg, or from about 950 mg to about 1000 mg.

[0195] The compounds described herein may be available in doses of approximately 1 mg, approximately 2 mg, approximately 3 mg, approximately 4 mg, approximately 5 mg, approximately 10 mg, approximately 15 mg, approximately 20 mg, approximately 25 mg, approximately 30 mg, approximately 35 mg, approximately 40 mg, approximately 45 mg, approximately 50 mg, approximately 55 mg, approximately 60 mg, approximately 65 mg, approximately 70 mg, approximately 75 mg, approximately 80 mg, approximately 85 mg, approximately 90 mg, approximately 95 mg, approximately 100 mg, approximately 125 mg, approximately 150 mg, approximately 175 mg, approximately 200 mg, approximately 250 mg, approximately 300 mg, approximately 350 mg, approximately 400 mg, approximately 450 mg, approximately 500 mg, approximately 550 mg, approximately 600 mg, approximately 650 mg, approximately 700 mg, approximately 750 mg, approximately 800 mg, approximately 850 mg, approximately 900 mg, approximately 950 mg, approximately 1000 mg. The composition is present in amounts of approximately 1050 mg, approximately 1100 mg, approximately 1150 mg, approximately 1200 mg, approximately 1250 mg, approximately 1300 mg, approximately 1350 mg, approximately 1400 mg, approximately 1450 mg, approximately 1500 mg, approximately 1550 mg, approximately 1600 mg, approximately 1650 mg, approximately 1700 mg, approximately 1750 mg, approximately 1800 mg, approximately 1850 mg, approximately 1900 mg, approximately 1950 mg, approximately 2000 mg, approximately 2100 mg, approximately 2200 mg, approximately 2300 mg, approximately 2400 mg, approximately 2500 mg, approximately 2600 mg, approximately 2700 mg, approximately 2800 mg, approximately 2900 mg, or approximately 3000 mg.

[0196] In some embodiments, the compound of this disclosure may be present in the composition in an amount from about 500 mg to about 2000 mg. In some embodiments, the compound of this disclosure may be present in the composition in an amount from about 1000 mg to about 1500 mg. In some embodiments, the compound of this disclosure may be present in the composition in an amount of about 500 mg. In some embodiments, the compound of this disclosure may be present in the composition in an amount of about 1000 mg. In some embodiments, the compound of this disclosure may be present in the composition in an amount of about 1500 mg. In some embodiments, the compound of this disclosure may be present in the composition in an amount of about 2000 mg.

[0197] In some embodiments, the dosage may be expressed as the amount of drug divided by the mass of the subject, for example, milligrams of drug per kilogram of the subject's body weight. In some embodiments, the compound is administered in amounts ranging from about 5 mg / kg to about 50 mg / kg, 250 mg / kg to about 2000 mg / kg, about 10 mg / kg to about 800 mg / kg, about 50 mg / kg to about 400 mg / kg, about 100 mg / kg to about 300 mg / kg, or about 150 mg / kg to about 200 mg / kg.

[0198] Companion diagnostic methods This document discloses a method for treating a condition in a subject with a corresponding need, comprising administering to the subject a therapeutically effective amount of a small molecule compound that binds to a p53 mutant protein, wherein the mutation status of the KRAS gene in the subject has been previously determined. In some embodiments, the mutation status of the KRAS gene in the subject has been previously determined based on assays performed on biological samples of the subject. In some embodiments, it has been previously determined that the subject has a mutation in the TP53 gene encoding a p53 mutant in the subject.

[0199] This document also discloses a method for treating a subject with a corresponding need, the method comprising: a) determining the mutation status of the KRAS gene in the subject based on the results of a assay performed on a biological sample of the subject; and b) administering to the subject a therapeutically effective amount of a compound binding to the p53 mutant protein, based at least in part on the mutation status of the KRAS gene in the subject. In some embodiments, the method further comprises determining, based on the results of the assay, that the subject carries a mutation in the TP53 gene encoding the p53 mutant protein, wherein the administration of a therapeutically effective amount of the compound binding to the p53 mutant protein to the subject is based at least in part on determining that the subject carries a mutation in the TP53 gene encoding the p53 mutant protein.

[0200] In some embodiments, the method further includes measuring a biological sample from the subject. In some embodiments, the method further includes measuring a biological sample from the subject, wherein measuring the biological sample from the subject further includes detecting a mutation in the TP53 gene encoding the p53 mutant in the subject.

[0201] In some embodiments, the mutation status of the KRAS gene in the subject includes the absence of missense mutations at any of codons 12 and 13. In some embodiments, the mutation status of the KRAS gene in the subject includes the absence of missense mutations at any of codons 12, 13, and 61. In some embodiments, the mutation status of the KRAS gene in the subject includes the absence of missense mutations at any of codons 12, 13, 59, 61, 117, and 146.

[0202] In some embodiments, the mutation status of the KRAS gene in the subject includes the absence of a mutation at codon 12 that encodes an amino acid other than glycine. In some embodiments, the mutation status of the KRAS gene in the subject includes the absence of a mutation at codon 13 that encodes an amino acid other than glycine. In some embodiments, the mutation status of the KRAS gene in the subject includes the absence of a mutation at codon 59 that encodes an amino acid other than alanine. In some embodiments, the mutation status of the KRAS gene in the subject includes the absence of a mutation at codon 61 that encodes an amino acid other than glutamine. In some embodiments, the mutation status of the KRAS gene in the subject includes the absence of a mutation at codon 117 that encodes an amino acid other than lysine. In some embodiments, the mutation status of the KRAS gene in the subject includes the absence of a mutation at codon 146 that encodes an amino acid other than alanine.

[0203] In some embodiments, the mutation status of the KRAS gene in the subject is the KRAS gene encoding the wild-type K-Ras protein. In some embodiments, the wild-type K-Ras protein has a sequence according to SEQ ID NO:1 or SEQ ID NO:2.

[0204] In some implementations, the mutation status of the KRAS gene in the subject is the copy number variation status of the KRAS gene. In some implementations, the mutation status of the KRAS gene in the subject includes the absence of KRAS copy number variation.

[0205] The mutation status of genes in subjects can be, for example, single nucleotide polymorphism (SNP), mutated gene sequence (MUT), loss of gene expression without gene mutation (LOE), wild-type gene sequence (WT), or complete gene deletion (NULL).

[0206] In some embodiments, the method of this disclosure further includes obtaining a biological sample from a subject. In some embodiments, the method of this disclosure further includes measuring the biological sample from the subject to detect mutations in genes encoding proteins (e.g., TP53, KRAS, PIK3CA, and BRCA1 / 2) in the subject.

[0207] In some embodiments, the mutation status of the BRCA1 or BRCA2 gene in the subject has been previously determined. In some embodiments, the mutation status of the BRCA1 or BRCA2 gene in the subject has been previously determined based on assays performed on biological samples of the subject. In some embodiments, it has been previously determined that the subject has a mutation in the TP53 gene, which encodes a p53 mutant in the subject.

[0208] This article also discloses a method for treating a subject with a corresponding need, the method comprising: administering to the subject a therapeutically effective amount of a compound binding to a p53 mutant protein, based at least in part on the mutation status of the BRCA1 or BRCA2 gene in the subject. This article also discloses a method for treating a subject with a corresponding need, the method comprising: a) determining the mutation status of the BRCA1 or BRCA2 gene in the subject; and b) administering to the subject a therapeutically effective amount of a compound binding to a p53 mutant protein, based at least in part on the mutation status of the BRCA1 or BRCA2 gene in the subject. This article also discloses a method for treating a subject with a corresponding need, the method comprising: a) determining the mutation status of the BRCA1 or BRCA2 gene in the subject based on the results of measurements performed on biological samples of the subject; and b) administering to the subject a therapeutically effective amount of a compound binding to a p53 mutant protein, based at least in part on the mutation status of the BRCA1 or BRCA2 gene in the subject. This document also discloses a method for treating a subject with a corresponding need, the method comprising: a) measuring a biological sample of the subject to determine the mutation status of the BRCA1 or BRCA2 gene in the subject; and b) administering to the subject a therapeutically effective amount of a compound binding to a p53 mutant protein, based at least in part on the mutation status of the BRCA1 or BRCA2 gene in the subject. In some embodiments, the method further includes determining, based on the results of the measurement, that the subject carries a mutation in the TP53 gene encoding the p53 mutant protein, wherein the administration of a therapeutically effective amount of the compound binding to the p53 mutant protein to the subject is based at least in part on determining that the subject carries a mutation in the TP53 gene encoding the p53 mutant protein.

[0209] This article also discloses a method for treating a subject with a corresponding need, the method comprising: administering to the subject a therapeutically effective amount of a compound binding to a p53 mutant protein, based at least in part on the mutation status of the PIK3CA gene in the subject. This article also discloses a method for treating a subject with a corresponding need, the method comprising: a) determining the mutation status of the PIK3CA gene in the subject; and b) administering to the subject a therapeutically effective amount of a compound binding to a p53 mutant protein, based at least in part on the mutation status of the PIK3CA gene in the subject. This article also discloses a method for treating a subject with a corresponding need, the method comprising: a) determining the mutation status of the PIK3CA gene in the subject based on the results of measurements performed on biological samples of the subject; and b) administering to the subject a therapeutically effective amount of a compound binding to a p53 mutant protein, based at least in part on the mutation status of the PIK3CA gene in the subject. This document also discloses a method for treating a subject with a corresponding need, the method comprising: a) measuring a biological sample of the subject to determine the mutation status of the PIK3CA gene in the subject; and b) administering to the subject a therapeutically effective amount of a compound binding to the p53 mutant protein, based at least in part on the mutation status of the PIK3CA gene in the subject. In some embodiments, the method further comprises determining, based on the results of the measurement, that the subject carries a mutation in the TP53 gene encoding the p53 mutant protein, wherein administering to the subject a therapeutically effective amount of the compound binding to the p53 mutant protein is based at least in part on determining that the subject carries a mutation in the TP53 gene encoding the p53 mutant protein.

[0210] In some embodiments, the homologous recombination deficiency (HRD) status of the cancer in the subject has been previously determined. In some embodiments, the HRD status of the cancer in the subject has been previously determined based on assays performed on biological samples of the subject. In some embodiments, it has been previously determined that the subject has a mutation in the TP53 gene encoding the p53 mutant in the subject.

[0211] This article also discloses a method for treating the condition of a subject in need, the method comprising: administering to the subject a therapeutically effective amount of a compound binding to a p53 mutant protein, based at least in part on the HRD status of the subject's cancer. This article also discloses a method for treating the condition of a subject in need, the method comprising: a) determining the HRD status of the subject's cancer; and b) administering to the subject a therapeutically effective amount of a compound binding to a p53 mutant protein, based at least in part on the HRD status of the subject's cancer. This article also discloses a method for treating the condition of a subject in need, the method comprising: a) determining the HRD status of the subject's cancer based on the results of measurements performed on the subject's biological samples; and b) administering to the subject a therapeutically effective amount of a compound binding to a p53 mutant protein, based at least in part on the HRD status of the subject's cancer. This article also discloses a method for treating the condition of a subject in need, the method comprising: a) measuring the subject's biological samples to determine the HRD status of the subject's cancer; and b) administering to the subject a therapeutically effective amount of a compound binding to a p53 mutant protein, based at least in part on the HRD status of the subject's cancer. In some implementations, the method further includes determining, based on the results of the assay, that the subject carries a mutation in the TP53 gene encoding the p53 mutant protein, wherein administration of a therapeutically effective amount of a compound binding the p53 mutant protein to the subject is based at least in part on determining that the subject carries a mutation in the TP53 gene encoding the p53 mutant protein.

[0212] In some embodiments, administering a therapeutically effective amount of the small molecule compound to the subject reduces the likelihood of p53 mutant degeneration in the subject. In some embodiments, administering a therapeutically effective amount of the small molecule compound to the subject reactivates the p53 pathway in the subject. In some embodiments, administering a therapeutically effective amount of the small molecule compound to the subject increases the anticancer activity of p53 mutants in the subject.

[0213] In some embodiments, the small molecule compound enhances the ability of the p53 mutant to bind to DNA. In some embodiments, the small molecule compound induces a conformational change in the p53 mutant. In some embodiments, the small molecule compound selectively binds to the p53 mutant compared to wild-type p53. In some embodiments, the small molecule compound increases the stability of the biologically active conformation of the p53 mutant relative to the stability of the biologically active conformation in the absence of the small molecule compound.

[0214] In some embodiments, mutations in the TP53 gene inactivate the p53 pathway in the subject. In some embodiments, the subject has cancer in which the cancer expresses a p53 mutant. In some embodiments, the mutation is at amino acid 220. In some embodiments, the p53 mutation is p53 Y220C. In some embodiments, the p53 mutant has a reduced ability to bind to DNA in the subject compared to wild-type p53. In some embodiments, the TP53 gene contains a frameshift mutation. In some embodiments, the TP53 gene contains a splice site mutation. In some embodiments, the TP53 gene contains an insertion mutation. In some embodiments, the TP53 gene contains a deletion mutation. In some embodiments, the TP53 gene contains a substitution mutation. In some embodiments, the TP53 gene contains a copy number variation. In some embodiments, the TP53 gene contains a copy number loss. In some embodiments, the TP53 gene contains a single nucleotide polymorphism. In some embodiments, the mutation in the TP53 gene is c.659A>G.

[0215] This document also discloses a method for treating a subject with a corresponding need, the method comprising: a) determining the mutation status of the KRAS gene in the subject based on the results of measurements performed on biological samples of the subject; and b) determining, at least in part, based on the mutation status of the KRAS gene in the subject, not to administer a compound that binds to the p53 mutant protein to the subject. In some embodiments, the mutation status of the KRAS gene in the subject includes a mutation at codon 12 encoding cysteine. In some embodiments, the mutation status of the KRAS gene in the subject includes a mutation at codon 12 encoding arginine. In some embodiments, the mutation status of the KRAS gene in the subject includes a mutation at codon 12 encoding serine. In some embodiments, the mutation status of the KRAS gene in the subject includes a mutation at codon 12 encoding alanine. In some embodiments, the mutation status of the KRAS gene in the subject includes a mutation at codon 12 encoding glutamate. In some embodiments, the mutation status of the KRAS gene in the subject includes a mutation at codon 12 encoding valine.

[0216] In some embodiments, this disclosure provides the use of the compounds herein (e.g., compound 1) in the treatment of cancer in a subject with a corresponding need, wherein the subject has been previously identified as lacking an oncogenic mutation in the KRAS gene based on assays performed on biological samples of the subject.

[0217] This document also discloses a method for selecting subjects for treatment with the compounds described herein (e.g., compound 1), the method comprising measuring a biological sample of the subject to determine the mutation status of the KRAS gene in the subject, wherein if the KRAS gene in the subject does not contain an oncogenic mutation, the subject is selected for treatment with the compounds described herein (e.g., compound 1). This document also discloses a method for selecting subjects for treatment with the compounds described herein, the method comprising determining the mutation status of the KRAS gene in the subject based on the results of measurements performed on a biological sample of the subject, wherein if the KRAS gene in the subject does not contain an oncogenic mutation, the subject is selected for treatment with the compounds described herein (e.g., compound 1).

[0218] In some implementations, the mutational status of the KRAS gene in the subject includes a mutation at codon 13 encoding aspartic acid.

[0219] In some embodiments, the mutation status of the KRAS gene in the subject includes a mutation at codon 59 encoding glutamate. In some embodiments, the mutation status of the KRAS gene in the subject includes a mutation at codon 59 encoding glycine. In some embodiments, the mutation status of the KRAS gene in the subject includes a mutation at codon 59 encoding threonine.

[0220] In some embodiments, the mutation status of the KRAS gene in the subject includes a mutation at codon 61 encoding lysine. In some embodiments, the mutation status of the KRAS gene in the subject includes a mutation at codon 61 encoding leucine. In some embodiments, the mutation status of the KRAS gene in the subject includes a mutation at codon 61 encoding arginine. In some embodiments, the mutation status of the KRAS gene in the subject includes a mutation at codon 61 encoding histidine.

[0221] In some implementations, the mutational status of the KRAS gene in the subject includes a mutation at codon 117 encoding asparagine.

[0222] In some embodiments, the mutation status of the KRAS gene in the subject includes a mutation at codon 146 encoding proline. In some embodiments, the mutation status of the KRAS gene in the subject includes a mutation at codon 146 encoding threonine. In some embodiments, the mutation status of the KRAS gene in the subject includes a mutation at codon 146 encoding valine.

[0223] In some embodiments, the mutation status of the KRAS gene in the subject is the KRAS gene encoding a mutant K-ras protein. In some embodiments, the mutant K-Ras protein does not have the sequence according to SEQ ID NO:1 or SEQ ID NO:2.

[0224] In some implementations, the mutation status of the KRAS gene in the subject is the copy number variation status of the KRAS gene. In some implementations, the mutation status of the KRAS gene in the subject is the presence of KRAS copy number variation.

[0225] This article also discloses a method for treating a subject with a corresponding need, the method comprising: at least in part based on the mutation status of the BRCA1 or BRCA2 gene in the subject, without administering to the subject a therapeutically effective amount of a compound binding to the p53 mutant protein. This article also discloses a method for treating a subject with a corresponding need, the method comprising: a) determining the mutation status of the BRCA1 or BRCA2 gene in the subject; and b) at least in part based on the mutation status of the BRCA1 or BRCA2 gene in the subject, without administering to the subject a therapeutically effective amount of a compound binding to the p53 mutant protein. This article also discloses a method for treating a subject with a corresponding need, the method comprising: a) determining the mutation status of the BRCA1 or BRCA2 gene in the subject based on the results of measurements performed on biological samples of the subject; and b) at least in part based on the mutation status of the BRCA1 or BRCA2 gene in the subject, without administering to the subject a therapeutically effective amount of a compound binding to the p53 mutant protein. This article also discloses a method for treating subjects in need of treatment, the method comprising: a) measuring a biological sample of the subject to determine the mutation status of the BRCA1 or BRCA2 gene in the subject; and b) not administering to the subject a therapeutically effective amount of a compound binding to the p53 mutant protein, based at least in part on the mutation status of the BRCA1 or BRCA2 gene in the subject.

[0226] In some embodiments, this disclosure provides the use of the compounds herein (e.g., compound 1) in treating cancer in a subject with a corresponding need, wherein the subject has previously been determined, based on assays performed on biological samples of the subject, to lack oncogenic mutations in the BRCA1 or BRCA2 genes.

[0227] This document also discloses a method for selecting a subject for treatment with a compound of this document (e.g., compound 1), the method comprising measuring a biological sample of the subject to determine the mutation status of the BRCA1 or BRCA2 gene in the subject, wherein if the BRCA1 or BRCA2 gene in the subject does not contain an oncogenic mutation, the subject is selected for treatment with a compound of this document (e.g., compound 1). This document also discloses a method for selecting a subject for treatment with a compound of this document (e.g., compound 1), the method comprising determining the mutation status of the BRCA1 or BRCA2 gene in the subject based on the results of measurements performed on a biological sample of the subject, wherein if the BRCA1 or BRCA2 gene in the subject does not contain an oncogenic mutation, the subject is selected for treatment with a compound of this document (e.g., compound 1).

[0228] This article also discloses a method for treating a subject in need of treatment, the method comprising: at least in part based on the mutation status of the PIK3CA gene in the subject, without administering to the subject a therapeutically effective amount of a compound binding to the p53 mutant protein. This article also discloses a method for treating a subject in need of treatment, the method comprising: a) determining the mutation status of the PIK3CA gene in the subject; and b) at least in part based on the mutation status of the PIK3CA gene in the subject, without administering to the subject a therapeutically effective amount of a compound binding to the p53 mutant protein. This article also discloses a method for treating a subject in need of treatment, the method comprising: a) determining the mutation status of the PIK3CA gene in the subject based on the results of measurements performed on biological samples of the subject; and b) at least in part based on the mutation status of the PIK3CA gene in the subject, without administering to the subject a therapeutically effective amount of a compound binding to the p53 mutant protein. This article also discloses a method for treating subjects in need of treatment, the method comprising: a) measuring a biological sample of the subject to determine the mutation status of the PIK3CA gene in the subject; and b) not administering to the subject a therapeutically effective amount of a compound binding to the p53 mutant protein, based at least in part on the mutation status of the PIK3CA gene in the subject.

[0229] In some embodiments, this disclosure provides the use of the compounds herein (e.g., compound 1) in treating cancer in a subject with a corresponding need, wherein the subject has previously been determined, based on assays performed on biological samples of the subject, to lack an oncogenic mutation in the PIK3CA gene.

[0230] In some embodiments, the mutation status of the PIK3CA gene in the subject is a PIK3CA gene that does not encode an oncogenic mutation. In some embodiments, the mutation status of the PIK3CA gene in the subject includes the absence of missense mutations at any one of codons 542, 545, 1047, and 1048. In some embodiments, the mutation status of the PIK3CA gene in the subject includes the absence of mutations at codon 542 that encode an amino acid other than glutamic acid. In some embodiments, the mutation status of the PIK3CA gene in the subject includes the absence of mutations at codon 545 that encode an amino acid other than glutamic acid. In some embodiments, the mutation status of the PIK3CA gene in the subject includes the absence of mutations at codon 1047 that encode an amino acid other than histidine. In some embodiments, the mutation status of the PIK3CA gene in the subject includes the absence of mutations at codon 117 that encode an amino acid other than lysine. In some embodiments, the mutation status of the PIK3CA gene in the subject is a PIK3CA gene encoding the wild-type p110α protein. In some implementations, the mutation status of the PIK3CA gene in the subject is the absence of copy number variation of the PIK3CA gene.

[0231] This document also discloses a method for selecting subjects to be treated with a compound of this document (e.g., compound 1), the method comprising measuring a biological sample of the subject to determine the mutation status of the PIK3CA gene in the subject, wherein if the PIK3CA gene in the subject does not contain an oncogenic mutation, the subject is selected to be treated with a compound of this document (e.g., compound 1). This document also discloses a method for selecting subjects to be treated with a compound of this document (e.g., compound 1), the method comprising determining the mutation status of the PIK3CA gene in the subject based on the results of measurements performed on a biological sample of the subject, wherein if the PIK3CA gene in the subject does not contain an oncogenic mutation, the subject is selected to be treated with a compound of this document (e.g., compound 1).

[0232] This article also discloses a method for treating the condition of a subject with a corresponding need, the method comprising: at least in part based on the HRD status of the subject's cancer, without administering to the subject a therapeutically effective amount of a compound binding to a p53 mutant protein. This article also discloses a method for treating the condition of a subject with a corresponding need, the method comprising: a) determining the HRD status of the subject's cancer; and b) at least in part based on the HRD status of the subject's cancer, without administering to the subject a therapeutically effective amount of a compound binding to a p53 mutant protein. This article also discloses a method for treating the condition of a subject with a corresponding need, the method comprising: a) determining the HRD status of the subject's cancer based on the results of measurements performed on the subject's biological samples; and b) at least in part based on the HRD status of the subject's cancer, without administering to the subject a therapeutically effective amount of a compound binding to a p53 mutant protein. This article also discloses a method for treating the condition of a subject with a corresponding need, the method comprising: a) measuring the subject's biological samples to determine the HRD status of the subject's cancer; and b) at least in part based on the HRD status of the subject's cancer, without administering to the subject a therapeutically effective amount of a compound binding to a p53 mutant protein.

[0233] In some embodiments, this disclosure provides the use of the compounds herein (e.g., compound 1) in the treatment of cancer in a subject with appropriate need, wherein the HRD status of the cancer has been previously determined based on assays performed on biological samples of the subject, wherein the HRD status is negative.

[0234] This document also discloses a method for selecting subjects to be treated with a compound of this document (e.g., compound 1), the method comprising measuring a biological sample of the subject to determine the HRD status of the subject's cancer, wherein if the HRD status of the cancer is negative, the subject is selected to be treated with a compound of this document (e.g., compound 1).

[0235] This article discloses a method for treating the condition of a subject in need, the method comprising administering to the subject a therapeutically effective amount of a compound of the following formula: Or its pharmaceutically acceptable salt.

[0236] Based on measurements of the subjects' biological samples, it has been determined that the subjects possess the TP53 gene encoding the Y220C p53 mutant protein and the KRAS gene encoding the wild-type K-ras protein.

[0237] In some implementations, it has been determined that the subject's biological sample contains mutations in the KRAS gene, which encodes the wild-type K-ras protein, and in the TP53 gene, which encodes the Y220C p53 mutant protein.

[0238] In some embodiments, this disclosure provides a method for treating the condition of a subject in need, the method comprising: a) Based on the results of measurements performed on the subjects' biological samples, i) Identify that the subject carries the KRAS gene encoding the wild-type K-ras protein, and ii) Identify that the subject carries a mutation in the TP53 gene, which encodes the Y220C p53 mutant protein; and b) at least in part based on i) Identify that the subject carries the KRAS gene encoding the wild-type K-ras protein, and ii) Identify whether the subject carries a mutation in the TP53 gene, which encodes the p53 mutant protein. Administer a therapeutically effective amount of the following compound to the subject: Or its pharmaceutically acceptable salt.

[0239] In some embodiments, the method further includes identifying whether the biological sample carries a mutation in the KRAS gene encoding the wild-type K-ras protein and a mutation in the TP53 gene encoding the Y220C p53 mutant protein.

[0240] In some embodiments, this disclosure provides a method comprising: (a) administering a dose of the compound provided herein (e.g., compound 1) to a subject, wherein the subject has cancer and is receiving a treatment regimen of the compound; (b) observing that the subject does not show improvement in cancer; (c) determining the mutation status of the KRAS gene in the subject, at least in part based on the observation that the subject does not show improvement in cancer; (d) determining that the mutation status of the KRAS gene in the subject is that the KRAS gene is non-wildtype; and (e) determining a treatment regimen to discontinue the compound, at least in part based on the mutation status of the KRAS gene. In some embodiments, the non-wildtype gene includes any mutations described herein.

[0241] In some implementations, this disclosure provides a treatment method in which recommendations are made based on observations of the patient's physiological state. Healthcare practitioners may recommend the administration of a therapeutic such as a pharmaceutical composition of this disclosure by, for example, consulting the subject or other healthcare provider, or by entering the recommendation into a medical record.

[0242] In some embodiments, this disclosure provides a method of treating a subject with cancer, the method comprising: (a) administering to the subject a first course of a compound binding to a p53 mutant; (b) after (a) determining that the subject has exhibited a gastrointestinal adverse event; (c) after (b) recommending that the compound be administered to the subject with food, wherein the recommendation is based at least on the determination that the subject has exhibited a gastrointestinal adverse event; and (d) administering to the subject a second course of the compound with food according to the recommendation.

[0243] In some embodiments, this disclosure provides a method of treating a subject with cancer, the method comprising: (a) administering to the subject a first course of a compound binding to a p53 mutant; (b) after (a) observing that the subject exhibits gastrointestinal adverse events; (c) after (b) determining, at least based on the observation that the subject has exhibited gastrointestinal adverse events, that the compound needs to be administered with food; and (d) according to the determination, administering to the subject a second course of the compound with food.

[0244] In some embodiments, this disclosure provides a method of treating cancer, the method comprising: (a) identifying a subject who: (i) has been treated with a first course of a compound binding to a p53 mutant, and (ii) has exhibited gastrointestinal adverse events following treatment with the first course of the compound; (b) recommending that the compound be administered to the subject with food, wherein the recommendation is based at least on the subject having exhibited gastrointestinal adverse events; and (c) administering a second course of the compound to the subject with food in accordance with the recommendation.

[0245] Clinical trials To determine the suitability of the compounds disclosed herein for human treatment, clinical trials are conducted. For example, patients diagnosed with cancer and requiring treatment are selected and divided into a treatment group and one or more subgroups, whereby the treatment group is administered the compounds disclosed herein, while the control group receives a placebo or a known anticancer drug. Therefore, the therapeutic safety and efficacy of the compounds disclosed herein can be evaluated by comparing patient groups regarding factors such as survival and quality of life. In this embodiment, the patient group treated with the compounds disclosed herein showed improved long-term survival compared to the control group treated with a placebo.

[0246] The results of genetic and biomarker tests, as well as additional tests on biological samples for biomarkers, can be investigated in relation to the safety and efficacy of the compounds disclosed herein, in relation to potential relevance to patient outcomes.

[0247] For example, clinical activity or clinical response can be evaluated using standard imaging assessments such as computed tomography (CT), magnetic resonance imaging (MRI), and bone scans. Additionally, positron emission tomography (PET) of [18F]-fluorodeoxyglucose and [18F]-fluorothymidine (FDG-PET and FLT-PET, respectively) or other techniques deemed clinically appropriate for the specific disease type of the patient can be used. For example, for DR-A, CT imaging can be performed at the end of cycle 2 and thereafter every two cycles (e.g., cycles 4 and 6), and in DR-B, CT imaging can be performed after the last infusion in cycle 3 and thereafter every three cycles (e.g., cycles 6 and 9). Anti-cancer cell activity can be assessed using the criteria for lymphoma patients outlined in IWG (2014) (Appendix H). Furthermore, for patients with FDG-avid lymphoma, FDG-PET imaging can be performed at baseline and post-baseline, as outlined in IWG 2014. For patients with cancerous cells, such as those with lymphoma, who typically show adequate uptake of the FLT tracer, FLT-PET imaging can be performed at baseline. For example, patients in the DR-A allocation who show a baseline standardized uptake value (SUV) >5 can have repeat FLT imaging performed one day after their last infusion of the study drug in cycle 1 (i.e., day 16). For example, patients in the DR-B allocation who show a baseline standardized uptake value (SUV) >5 can have repeat FLT imaging performed one day after their last infusion of the study drug in cycle 1 (i.e., day 12).

[0248] biological samples Biological samples can be fluids or other materials derived from the body of a normal or diseased subject, such as blood, serum, plasma, lymph, urine, saliva, tears, cerebrospinal fluid, breast milk, amniotic fluid, bile, ascites, pus, and the like. In some embodiments, biological samples are organ or tissue extracts and culture media in which any cell or tissue product from the subject has been incubated. Biological samples can be any sample from which genetic material can be obtained. Biological samples can also include solid or liquid cancer cell samples or specimens. Cancer cell samples can be cancer cell tissue samples. Cancer cell samples can be primary cancer samples, metastatic cancer samples, or recurrent cancer samples. In some embodiments, cancer cell tissue samples can be obtained from surgically removed tissue. Exemplary sources of biological samples include fine-needle aspiration, core needle biopsy, vacuum-assisted biopsy, incision biopsy, excisional biopsy, punch biopsy, shave biopsy, or skin biopsy. In some cases, biological samples include fine-needle aspiration samples. In some embodiments, the biological sample includes tissue samples, including, for example, excisional biopsies, incisional biopsies, or other biopsies. The biological sample may comprise a mixture of two or more sources; for example, fine-needle aspiration and tissue samples. Tissue and cell samples can also be obtained without invasive surgery, such as by puncturing the chest or abdominal wall with a fine needle or by aspirating cellular material from a mass in the breast, thyroid, or other sites (fine-needle aspiration biopsy). In some embodiments, the biological sample is a bone marrow aspiration sample. Biological samples can be obtained by methods known in the art, such as the biopsy methods provided herein, swabbing, curettage, venotomy, or any other suitable method.

[0249] The obtained biological samples can be used fresh, frozen, or fixed (e.g., formaldehyde-fixed paraffin-embedded), depending on the nature of the sample, the assay used, and the practitioner's convenience. Although fresh, frozen, and fixed materials are suitable for a variety of RNA and protein assays, fresh tissue is generally preferred for in vitro measurements of activity.

[0250] Fixed tissue samples can also be used. Tissue obtained through biopsy is typically fixed, usually by means of formalin, formaldehyde, or glutaraldehyde, or by immersion in alcohol. Fixed biological samples are typically dehydrated and embedded in paraffin or other solid supports. Non-embedded fixed tissue as well as fixed and embedded tissue can be used in the methods of the present invention. The solid support used to embed the fixed tissue can be removed with an organic solvent to allow subsequent rehydration of the preserved tissue.

[0251] In some cases, the assays described herein include steps of cell culture or tissue culture. For example, cells from a biopsy may be depolymerized using enzymes (such as collagenase and hyaluronidase) and / or physical disruption (e.g., repeated passage through a 25-gauge needle) to dissociate the cells, collected by centrifugation, and resuspended in the desired buffer or culture medium for culture, immediate analysis, or further processing.

[0252] Subjects / Patients In some embodiments, the subjects treated with the methods provided herein are humans who already have or have been diagnosed with cancer. In other embodiments, the subjects treated with the methods provided herein are humans who are predisposed to or susceptible to developing cancer. In some embodiments, the subjects treated with the methods provided herein are humans at risk of developing cancer.

[0253] In some embodiments, the subjects treating cancer according to the methods provided herein are humans who already have or have been diagnosed with cancer identified as carrying a p53 inactivating mutation and / or lacking wild-type p53. In other embodiments, the subjects treating cancer according to the methods provided herein are humans who are predisposed to or susceptible to cancer, identified as carrying a p53 inactivating mutation and / or lacking wild-type p53. In some embodiments, the subjects treating cancer using the methods provided herein are humans at risk of developing cancer, identified as carrying a p53 inactivating mutation and / or lacking wild-type p53.

[0254] p53 inactivation mutations are mutations that result in the loss (or reduction) of p53's apoptotic activity in vitro. Non-restrictive examples of p53 inactivation mutations include mutations at Val143, His168, Arg175, Tyr220, Gly245, Arg248, Arg249, Phe270, Arg273, and Arg282. Examples include mutations at Y220C, Y220S, Y220H, R248Q, R248W, R273C, and R273H. Other non-restrictive examples of p53 inactivation mutations include E62_W91del, V122X, C135S, V143A, Q144P, W146X, V157F, R158H, Y163N, H168Y, V173L, R175H, R175L, R175P, R175Q, R175S, P219H, Y234C, Y234H, and M237I. S240R, G245C, G245S, M246I, R249S, I255N, V272M, R273L, V274F, G279E, R280K, D281H, R282W, R306P, P300_L308del, P300_Y327del, D324_I332del, R337C, and L344P, where X is any amino acid.

[0255] Therefore, in some implementations, subjects who use the methods provided herein to treat cancer are humans who are at risk of developing cancer carrying a p53 inactivating mutation, and who already have or have been diagnosed with the cancer.

[0256] In some implementations, the subjects treated with cancer according to the methods provided herein are humans who already have or have been diagnosed with cancer identified as lacking a dominant p53 inactivating mutation. As used herein, a dominant p53 inactivating mutation or a dominant negative mutation is a mutation in which the mutated p53 suppresses or disrupts the activity of the wild-type p53 gene.

[0257] In some embodiments, the subject treating cancer according to the methods provided herein is a treatment-resistant patient. In some embodiments, a treatment-resistant patient is one who is resistant to standard therapies (e.g., surgery, radiation, anti-androgen therapy, and / or drug therapies such as chemotherapy). In some embodiments, a patient with cancer is treatment-resistant when the cancer is not significantly eradicated and / or one or more symptoms are not significantly relieved. Whether a patient is treatment-resistant can be determined in vivo or in vitro by any effective method for assessing the effectiveness of cancer treatment. In various embodiments, a patient with cancer is treatment-resistant when the number of cancer-associated CTCs or MNBCs has not decreased or has increased. In various embodiments, a patient with cancer is treatment-resistant when one or more types of cancer cells have metastasized and / or spread to another organ.

[0258] In some embodiments, subjects treated with the methods provided herein are humans who have been shown to be refractory to therapies other than those disclosed herein but are no longer receiving such therapies. In some embodiments, subjects treated with the methods provided herein are humans who have received one or more conventional anticancer therapies, such as surgery, drug therapies such as chemotherapy, anti-androgen therapy, or radiation. These patients include refractory patients, patients who are too young for conventional therapies, and patients who have recurrent cancer cells despite existing treatments.

[0259] In some implementations, the subjects are humans who have already undergone at least one unsuccessful prior cancer treatment and / or cancer therapy.

[0260] Methods for detecting wild-type proteins and / or mutations In some embodiments, subjects with cancer that does not contain a KRAS gene with an oncogenic mutation are candidates for cancer treatment with the compounds of this disclosure. In some embodiments, the cancer contains a p53 inactivating mutation. Prior to treatment with the compounds of this disclosure, cancer cells from the patient group can be assayed to determine mutations and / or expression of wild-type proteins (e.g., p53 and / or K-ras).

[0261] In some embodiments, subjects selected for treatment with the p53-activating compound of this disclosure are selected at least in part based on the mutation status of one or more genes. In cases where subjects are selected for treatment based on the mutation status of more than one gene, selection may be based at least in part on the results of a single assay or more than one assay. For example, a single assay may be performed on a biological sample of the subject (e.g., a solid tumor sample) to provide results describing the mutation status of two or more genes (e.g., TP53, KRAS, BRCA1, and / or BRCA2).

[0262] In some embodiments, the assay is DNA sequencing. In some embodiments, the assay is next-generation DNA sequencing. In some embodiments, the assay is RNA sequencing. In some embodiments, the assay is real-time PCR.

[0263] In some embodiments, the biological sample is a liquid biopsy. In some embodiments, the biological sample is a blood sample. In some embodiments, the biological sample is circulating tumor DNA. In some embodiments, the biological sample is cell-free DNA. In some embodiments, the biological sample is a solid tumor sample.

[0264] In some implementations, the choice of treatment is based at least in part on the results of more than one assay. The assays may be of the same type or different types, and may be performed simultaneously or sequentially by the same or different parties. Each assay may be performed on the same biological sample or different biological samples. For example, a subject's biological sample may be subjected to an assay to determine the mutation status of at least one gene, optionally stored for a period of time, and another assay to determine the mutation status of another gene may be performed. Optionally or additionally, a first biological sample (e.g., a solid tumor sample) may be subjected to an assay to determine the mutation status of one gene (e.g., TP53), and a second biological sample (e.g., circulating tumor DNA) may be subjected to a second assay to determine the status of another gene (e.g., KRAS, BRCA1, and / or BRCA2).

[0265] The activity of the p53 pathway can be determined by the mutation status of genes involved in the p53 pathway. Genes involved in the p53 pathway include, for example, AKT1, AKT2, AKT3, ALK, BRAF, CDK4, CDKN2A, DDR2, EGFR, ERBB2 (HER2), FGFR1, FGFR3, GNA11, GNQ, GNAS, KDR, KIT, KRAS, MAP2K1 (MEK1), MET, HRAS, NOTCH1, NRAS, NTRK2, PIK3CA, NF1, PTEN, RAC1, RB1, NTRK3, STK11, PIK3R1, TSC1, TSC2, RET, TP53, and VHL. It can also assess genes that regulate p53 activity, including, for example, kinases: ABL1, JAK1, JAK2, JAK3; receptor tyrosine kinases: FLT3 and KIT; receptors: CSF3R, IL7R, MPL, and NOTCH1; transcription factors: BCOR, CEBPA, CREBBP, ETV6, GATA1, GATA2, MLL, KZF1, PAX5, RUNX1, STAT3, WT1, and TP53; epigenetic factors: ASXL1, DNMT3A, EZH2, KDM6A(UTX), SUZ12, TET2, PTPN11, SF3B1, SRSF2, U2AF35, and ZRSR2; RAS proteins: HRAS, KRAS, and NRAS; adaptors CBL and CBL-B; FBXW7, IDH1, IDH2, and NPM1.

[0266] Cancer cell samples can be obtained from solid tumors or liquid tumors, for example, via resection of primary or metastatic tumors (e.g., lung resection, lobectomy, wedge resection, and craniotomy), biopsy of primary or metastatic disease (e.g., transbronchial or needle core), pleural fluid or ascites (e.g., FFPE cell deposits), bone marrow aspirate, bone marrow clots and bone marrow biopsy, or macroscopic anatomy of tumor-rich areas (solid tumors).

[0267] To detect mutational status of KRAS, TP53, BRCA1, and / or BRCA2 in tissue, cancerous tissue can be isolated from surrounding normal tissue. For example, tissue can be isolated from paraffin or frozen sections. Cancer cells can also be separated from normal cells by flow cytometry. Detection of mutations may be more difficult if cancerous tissue is highly contaminated with normal cells.

[0268] A variety of methods and assays for analyzing mutations are suitable for use in this disclosure. Non-limiting examples of assays include polymerase chain reaction (PCR), quantitative PCR (qPCR), real-time PCR (RT-PCR), Sanger sequencing, restriction fragment length polymorphism (RFLP), microarrays, Southern blotting, Northern blotting, Western blotting, Eastern blotting, H&E staining, microscopic evaluation of tumors, massively parallel sequencing (MPS), next-generation sequencing (NGS) (e.g., DNA extraction, purification, quantification, and amplification, library preparation), immunohistochemistry (IHC), protein quantification, chromogenic in situ hybridization (CISH), and fluorescence in situ hybridization (FISH).

[0269] Microarrays allow researchers to study multiple DNA sequences attached to a surface, such as a DNA chip made of glass or silicon, or polymer beads or resin. The DNA sequences hybridize with fluorescent or luminescent probes. Microarrays can indicate the presence of oligonucleotide sequences in a sample based on the hybridization of the sample sequence with the probe, followed by washing and subsequent detection of the probe. Quantification of the fluorescent or luminescent signal indicates the presence of a known oligonucleotide sequence in the sample.

[0270] PCR allows for the rapid amplification of DNA oligomers and can be used to identify oligonucleotide sequences in a sample. A PCR experiment involves contacting an oligonucleotide sample with a PCR mixture containing primers complementary to the target sequence, one or more DNA polymerases, deoxyribonucleotide triphosphate (dNTP) structural units (including dATP, dGTP, dTTP, and dCTP), suitable buffers, salts, and additives. If the sample contains an oligonucleotide sequence complementary to a pair of primers, the experiment amplifies the sample sequence, which can then be collected and identified.

[0271] In some embodiments, the assay includes amplifying a biomolecule from a cancer sample. The biomolecule can be a nucleic acid molecule, such as DNA or RNA. In some embodiments, the assay includes circularization of the nucleic acid molecule, followed by digestion of the circularized nucleic acid molecule.

[0272] In some embodiments, the assay involves contacting an organism or a biochemical sample collected from an organism (such as a nucleic acid sample) with a library of oligonucleotides (such as PCR primers). The library may contain any number of oligonucleotide molecules. Oligonucleotide molecules may bind individual DNA or RNA motifs, or any combination of motifs described herein. Motifs may be spaced at any distance, and the distance may be known or unknown. In some embodiments, two or more oligonucleotides in the same library bind to motifs spaced at known distances within the parental nucleic acid sequence. Primer binding to the parental sequence may occur based on the complementarity of the primer and parental sequence. Binding may occur, for example, under annealing or under stringent conditions.

[0273] In some embodiments, the results of the assay are used to design new oligonucleotide sequences for future use. In some embodiments, the results of the assay are used to design new oligonucleotide libraries for future use. In some embodiments, the results of the assay are used to revise, improve, or update existing oligonucleotide libraries for future use. For example, the assay may reveal that previously undocumented nucleic acid sequences are associated with the presence of target material. This information can be used to design or redesign nucleic acid molecules and libraries.

[0274] In some embodiments, one or more nucleic acid molecules in the library contain barcode tags. In some embodiments, one or more nucleic acid molecules in the library contain type I or type II restriction sites suitable for circularizing and cleaving amplified sample nucleic acid sequences. Such primers can be used to circularize PCR products and cleave PCR products to provide product nucleic acid sequences that are constructed differently from the untreated nucleic acid sequences of the sample organism.

[0275] Following a PCR experiment, the presence of the amplified sequence can be verified. Non-limiting examples of methods for identifying amplified sequences include DNA sequencing, whole transcriptome shotgun sequencing (WTSS or RNA-seq), mass spectrometry (MS), microarrays, pyrosequencing, column purification analysis, polyacrylamide gel electrophoresis, and index-tagged sequencing of PCR products generated from indexed primers.

[0276] In some implementations, more than one nucleic acid sequence is amplified from the organic tissue of the sample. Non-limiting examples of methods for isolating different nucleic acid sequences from a mixture of PCR products include column purification, high-performance liquid chromatography (HPLC), HPLC / MS, polyacrylamide gel electrophoresis, and size exclusion chromatography.

[0277] Amplified nucleic acid molecules can be identified by sequencing. Nucleic acid sequencing can be performed on automated instruments. Sequencing experiments can be performed in parallel to analyze dozens, hundreds, or thousands of sequences simultaneously. Non-limiting examples of sequencing technologies are as follows.

[0278] In pyrosequencing, DNA is amplified within water droplets in an oil solution, each droplet containing a single DNA template bound to primer-coated beads. Nucleotides are added to the grown sequence, and the addition of each base is verified by visible light.

[0279] Ion semiconductor sequencing detects the addition of nucleic acid residues as an electrical signal associated with hydrogen ions released during synthesis. Reaction wells containing the template are filled with four types of nucleotide structural units, one at a time. The timing of the electrical signal identifies which structural unit was added and the corresponding residue in the template.

[0280] DNA nanospheres amplify DNA into nanospheres using rolling circle replication. Non-stranded sequencing performed through nanosphere linking reveals the DNA sequence.

[0281] In the reversible dye method, nucleic acid molecules are annealed to primers on a glass slide and amplified. Four types of fluorescent dye residues, each complementary to an untreated nucleobase, are added. Residues complementary to the next base in the nucleic acid sequence are then added, and unincorporated dye is washed off the slide. Four types of reversible terminator bases (RT-bases) are added, and unincorporated nucleotides are washed away. The addition of fluorescent indicator dye residues identifies complementary bases in the template sequence. The dye residues are then chemically removed, and the cycle is repeated.

[0282] Detection of point mutations can be accomplished by molecularly cloning and sequencing the KRAS, PIK3CA, TP53, BRCA1, and / or BRCA2 alleles present in cancer cell tissue. Alternatively, PCR can be used to amplify the KRAS, PIK3CA, TP53, BRCA1, and / or BRCA2 gene sequences directly from genomic DNA artifacts from cancer cell tissue. The amplified DNA sequence can then be determined. Specific deletions of the KRAS, PIK3CA, TP53, BRCA1, and / or BRCA2 genes can also be detected. For example, restriction fragment length polymorphism (RFLP) probes targeting the KRAS, PIK3CA, TP53, BRCA1, and / or BRCA2 genes or surrounding marker genes can be used to score the loss of KRAS, TP53, PIK3CA, BRCA1, and / or BRCA2 alleles.

[0283] The loss of wild-type KRAS, TP53, PIK3CA, BRCA1, and / or BRCA2 genes can also be detected based on the loss of wild-type expression products of these genes. Such expression products include mRNA and p53, BRCA1, BRCA2, and / or K-ras protein products. Point mutations can be detected by direct sequencing of mRNA or by molecular cloning of cDNA prepared from mRNA. The sequence of the cloned cDNA can be determined using DNA sequencing technology. cDNA can also be sequenced by PCR.

[0284] Alternatively, mismatch detection can be used to detect point mutations in the KRAS, TP53, PIK3CA, BRCA1, and / or BRCA2 genes or mRNA products. The method may involve using a labeled riboprobe complementary to the human wild-type KRAS, TP53, PIK3CA, BRCA1, and / or BRCA2 genes. The riboprobe and mRNA or DNA isolated from cancer cell tissue are annealed (hybridized) together and subsequently digested with RNase A, an enzyme capable of detecting some mismatches in the double-stranded RNA structure. If a mismatch is detected by RNase A, the enzyme cleaves at the mismatch site. Therefore, when the annealed RNA product is separated on an electrophoresis gel matrix, if a mismatch has been detected and cleaved by RNase A, the RNA product is seen as a full-length double-stranded RNA smaller than the riboprobe and the KRAS, TP53, PIK3CA, BRCA1, and / or BRCA2 mRNA or DNA. The RNA probe does not need to be the full length of KRAS, TP53, PIK3CA, BRCA1, and / or BRCA2 mRNA or gene, but can be a segment of any of them. For example, if the RNA probe only contains segments of KRAS, TP53, PIK3CA, BRCA1, and / or BRCA2 mRNA or gene, many of these probes can be used to screen for mismatches in the entire mRNA sequence.

[0285] In a similar manner, DNA probes can be used to detect mismatches via enzymatic or chemical cleavage. Alternatively, mismatches can be detected by observing changes in the electrophoretic mobility of the mismatched duplex relative to the matched duplex. Using ribonucleic acid or DNA probes, PCR (see below) can be used to amplify cellular mRNA or DNA that may contain mutations prior to hybridization.

[0286] Allele-specific probes can also be used to screen DNA sequences of the KRAS, TP53, PIK3CA, BRCA1, and / or BRCA2 genes that have been amplified by PCR from cancer cell tissue. These probes are nucleic acid oligomers, each containing a region of the KRAS, TP53, PIK3CA, BRCA1, and / or BRCA2 gene sequence carrying a known mutation. For example, an oligomer may be about 30 nucleotides long, corresponding to a portion of the KRAS, TP53, PIK3CA, BRCA1, and / or BRCA2 gene sequence. At codon 175 of the p53 gene, the oligomer encodes alanine instead of the wild-type codon valine. By using a set of such allele-specific probes, PCR amplification products can be screened to identify the presence of previously identified mutations in the KRAS, TP53, PIK3CA, BRCA1, and / or BRCA2 genes. Hybridization of allele-specific probes with amplified KRAS, TP53, PIK3CA, BRCA1, and / or BRCA2 sequences can be performed, for example, on a nylon filter. Hybridization with a specific probe indicates the presence of the same mutation in the allele-specific probe within cancer cell tissue.

[0287] The identification of structural changes in KRAS, TP53, PIK3CA, BRCA1, and / or BRCA2 genes in cancer cells can be facilitated by applying a range of different high-resolution, high-throughput microarray platforms. Essentially, two types of arrays exist: arrays carrying PCR products of nucleic acids from clones (e.g., cDNA, BAC, viscera) and arrays using oligonucleotides. Methods can provide ways to investigate genome-wide DNA copy number aberrations and expression levels, allowing for associations between loss, gain, and amplification in cancer cells and genes overexpressed and underexpressed in the same sample. Gene expression arrays providing estimates of mRNA levels in cancer cells have been developed, resulting in exon-specific arrays capable of identifying gene expression levels, selective splicing events, and mRNA processing alterations.

[0288] Oligonucleotide arrays can be used to query single nucleotide polymorphisms (SNPs) throughout the genome for linkage and association studies, and these have been applied to quantify copy number abnormalities and loss of heterozygosity events. DNA sequencing arrays can allow for the resequencing of chromosomal regions and the entire genome.

[0289] SNP-based arrays or other gene arrays or microarrays can determine the presence and structure of wild-type TP53 and / or KRAS alleles. Single nucleotide polymorphisms (SNPs), variations at a single site in DNA, are the most common type of variation in the genome. For example, it is estimated that there are 5 to 10 million SNPs in the human genome. SNPs can be synonymous or nonsynonymous substitutions. Due to the degeneracy of the genetic code, synonymous SNP substitutions do not lead to changes in the amino acids of proteins, but can affect function in other ways. For example, a seemingly silent mutation in a gene encoding a membrane transporter can slow translation, allow peptide misfolding, and produce a weaker mutant membrane transporter. Nonsynonymous SNP substitutions can be missense or nonsense substitutions. Missense substitutions occur when a single base change leads to a change in the amino acid sequence of a protein and its dysfunction results in disease. Nonsense substitutions occur when point mutations result in premature stop codons or nonsense codons in transcribed mRNA, leading to truncated and usually nonfunctional protein products. Because SNPs are highly conserved throughout evolution and within populations, SNP maps serve as excellent genotypic markers for research. SNP arrays are a useful tool for studying the entire genome.

[0290] Furthermore, SNP arrays can be used to study loss of heterozygosity (LOH). LOH is a form of allelic imbalance that can be caused by the complete loss of an allele or by an increase in the copy number of one allele relative to another. While other microarray-based methods exist (e.g., comparative genomic hybridization can only detect genomic gains or deletions), SNP arrays have the added advantage of detecting copy number-neutral LOH caused by uniparental disomy (UPD). In UPD, the deletion of one allele or an entire chromosome from one parent results in a duplicated copy of the allele from the other parent (uniparent = from one parent, disomy = duplicated). In disease settings, this condition can be pathological when the wild-type allele (e.g., from the mother) is absent and two copies of a heterozygous allele (e.g., from the father) are present. This use of SNP arrays has great potential in cancer diagnosis, as LOH is a prominent feature of most human cancers. SNP array technology has been shown to have a high rate of LOH (allelic imbalance) in cancers (e.g., gastric cancer, liver cancer, etc.) and hematologic malignancies (ALL, MDS, CML, etc.) due to genomic deletions or UPD and genomic gains. In this disclosure, the detection of LOH using high-density SNP arrays allows for the identification of patterns of allelic imbalance to determine the presence of the wild-type p53 allele.

[0291] Examples of p53 gene sequences and SNP arrays include the p53 Gene Chip (Affymetrix, Santa Clara, CA) and the Ampli-Chip.® p53 microarray (Roche Molecular Systems, Pleasanton, CA), GeneChipMapping array (Affymetrix, Santa Clara, CA), SNP Array 6.0 (Affymetrix, Santa Clara, CA), BeadArrays (Illumina, San Diego, CA), etc.

[0292] Non-limiting examples suitable for determining KRAS mutation status include therascreen KRAS RGQPCR kit (QIAGEN, Hilden, Germany), KRAS XL StripAssay® (ViennaLab Diagnostics, Wien, Austria), SNaPshot multiplex PCR, cobas® KRAS mutation assay (Roche Diagnostics, FlorhamPark, NJ), and ddPCR™. KRAS G12 / G13 Screening kits (Bio-Rad Laboratories, Philadelphia, PA), OncoBEAM® RAS CRC kits (Sysmex Inostics, Baltimore, MD), and Idylla TM KRAS mutation test (Biocartis, Mechelen, Belgium).

[0293] Non-limiting examples of assays suitable for determining BRCA1 / 2 mutation status include the BRACANALYSIS CDX® germline companion diagnostic test (Myriad Genetics, Salt Lake City, UT), the CleanPlex® BRCA1 & BRCA2 kit v3 (Paragon Genomics, Inc., Fremont, CA), the Guardant360® CDx (Guardant, Palo Alto, CA), and the BRCAssure® (Laboratory Corporation of America, Burlington, NC).

[0294] Non-limiting examples of assays suitable for determining PIK3CA mutation status include the Agena PIK3CA Breast Group (Agena Bioscience, San Diego, CA) and the Invitae PIK3CA Diagnostics (Invitae, San Francisco, CA). therascreen PIK3CA RGQ PCR Kit (QIAGEN, Hilden, Germany), PIK3CA Mutation Analysis Kit for Real-Time PCR (EnteroGen, Woodland Hills, CA), FoundationOne® CDx (Cambridge, MA), Caris® Life Science Caris Molecular Intelligence® Tumor Profiling Analysis (Irving, TX), Illumina® TruSight TM Oncology 500 (San Diego, CA), NeoGenomics NeoTYPE® (Fort Meyers, FL), Tempus xT (Chicago, IL), and Thermo Fisher Oncomine TM Dx target test (Waltham, MA).

[0295] Mutations in wild-type KRAS, TP53, PIK3CA, BRCA1, and / or BRCA2 genes can also be detected based on mutations in the wild-type expression products of the KRAS, TP53, PIK3CA, BRCA1, and / or BRCA2 genes. Such expression products include mRNA as well as the p110α, p53, BRCA1, BRCA2, and / or K-ras protein products themselves. Point mutations can be detected by direct sequencing of the mRNA or via molecular cloning of cDNA prepared from the mRNA. The sequence of the cloned cDNA can be determined using DNA sequencing technology. The cDNA can also be sequenced via polymerase chain reaction (PCR). A set of monoclonal antibodies can be used, where each epitope involved in the function of p53, BRCA1, BRCA2, and / or K-ras is represented by a monoclonal antibody. Loss or perturbation of binding to the monoclonal antibodies in this set can indicate mutational changes in the p53 and / or K-ras proteins, and therefore indicate mutational changes in the KRAS, TP53, PIK3CA, BRCA1, and / or BRCA2 genes themselves. Mutant TP53 and / or KRAS genes or their products can also be detected in bodily samples, including, for example, serum, feces, urine, and sputum. The same techniques discussed above for detecting mutant KRAS, TP53, PIK3CA, BRCA1, and / or BRCA2 genes or their products in tissues can be applied to other bodily samples.

[0296] Loss of wild-type KRAS, TP53, PIK3CA, BRCA1, BRCA2, and / or K-ras proteins can also be detected by screening for loss of function of wild-type p53, PIK3CA, BRCA1, and / or BRCA2 genes. While not all functions of the p53 protein have been elucidated, at least two specific functions are known. The p53 protein binds to the SV40 large T antigen and the adenovirus E1B antigen. Loss of the ability of the p53 protein to bind to any one or both of these antigens indicates a mutational change in the protein, which reflects a mutational change in the gene itself. Alternatively, a set of monoclonal antibodies can be used, where each epitope involved in p53 function is represented by a monoclonal antibody. Loss or perturbation of binding to monoclonal antibodies in this set indicates a mutational change in the p53 protein, and therefore indicates a mutational change in the p53 gene itself. Any method used to detect altered p53 protein can be used to detect loss of the wild-type p53 gene.

[0297] The determination of p53 inactivation mutations and / or the absence or reduced expression of wild-type p53 in subjects with cancer can be performed before, during, or after administration of the compounds disclosed herein. The determination of KRAS mutations and / or the absence of wild-type K-ras expression in subjects with cancer can be performed before, during, or after administration of the compounds disclosed herein. The determination of BRCA1 and / or BRCA2 mutations and / or the absence of wild-type BRCA1 and / or BRCA2 expression in subjects with cancer can be performed before, during, or after administration of the compounds disclosed herein. The determination of PIK3CA mutations and / or the absence of wild-type p110α expression in subjects with cancer can be performed before, during, or after administration of the compounds disclosed herein. In some implementations, the absence of the KRAS mutation and / or the determination of wild-type K-ras expression are performed prior to the first administration of the compound to the subject, for example, approximately 5 years to approximately 1 month, approximately 4 years to approximately 1 month, approximately 3 years to approximately 1 month, approximately 2 years to approximately 1 month, approximately 1 year to approximately 1 month, approximately 5 years to approximately 1 week, approximately 4 years to approximately 1 week, approximately 3 years to approximately 1 month, approximately 2 years to approximately 1 week, approximately 1 year to approximately 1 week, approximately 5 years prior to the first administration of the compound to the subject. One year - about 1 day, about 4 years - about 1 day, about 3 years - about 1 day, about 2 years - about 1 day, about 1 year - about 1 day, about 15 months - about 1 month, about 15 months - about 1 week, about 15 months - about 1 day, about 12 months - about 1 month, about 12 months - about 1 week, about 12 months - about 1 day, about 6 months - about 1 month, about 6 months - about 1 week, about 6 months - about 1 day, about 3 months - about 1 month, about 3 months - about 1 week or about 3 months - about 1 day. In some instances, confirmation of KRAS mutation deficiency and / or wild-type K-ras expression is performed up to 6 years, 5 years, 4 years, 3 years, 24 months, 23 months, 22 months, 21 months, 20 months, 19 months, 18 months, 17 months, 16 months, 15 months, 14 months, 13 months, 12 months, 11 months, 10 months, 9 months, 8 months, 7 months, 6 months, 5 months, 4 months, 3 months, 2 months, 1 month, 4 weeks (28 days), 3 weeks (21 days), 2 weeks (14 days), 1 week (7 days), 6 days, 5 days, 4 days, 3 days, 2 days, or 1 day prior to the first administration of the compound to the subject. In some embodiments, confirmation of KRAS mutation deficiency is performed within 1 month of the first administration of the compound to the subject. In some embodiments, confirmation of KRAS mutation deficiency is performed within 21 days of the first administration of the compound to the subject.

[0298] Example Example 1: Retrospective confirmatory test for p53 Y220C mutation.

[0299] NGS is used for retrospective confirmatory testing for p53 Y220C mutations in blood or solid tissues. If a patient does not have the p53 Y220C mutation detected in the pre-screening step, the patient can be prospectively recruited.

[0300] DNA was obtained from a cancer sample from a patient. NGS sequencing was used to sequence the DNA to determine the entire p53 coding region, including TP53 exons, introns, and splice sites. Dysfunctional p53 was inferred from the identification of substitutions, insertions / deletions, frameshift mutations, splice site mutations, insertions or deletions, copy number variants, large deletions, or polymorphisms. The minimum tumor content was 20%. The average read depth was approximately 750 reads per amplicon. In each amplicon... > With an average read depth of 450 reads, the detection limit is 5% for mutant alleles. When the average read depth is <450 reads per amplicon, the detection limit is 15% for mutant alleles.

[0301] TP53 gene copy number determination is based on comparing the number of TP53 amplicon reads from tumors to the average number of reads from 14 normal DNA samples. Limitations of the assay: If tumor content >60% (99% sensitivity), one or more allele losses are identified; if tumor sample content >30% (99% sensitivity), two allele losses are identified. More than 10% of tumors possessing wild-type TP53 have a copy number less than 0.5 (<0.5).

[0302] A regimen of administering a compound containing a therapeutically effective amount of the compound of this disclosure or a pharmaceutically acceptable salt thereof to a patient with a confirmed Y220C mutation.

[0303] Example 2: Phase 1 clinical trial in patients with advanced solid tumors carrying the p53 Y220C mutation Compound 1 (4-((3-(4-((((3S,4R)-3-fluoro-1-methylpiperidin-4-yl)amino)-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl)amino)-3-methoxy-N-methylbenzamide) is a selective p53 reactivator that stabilizes the p53 Y220C mutant protein. A single amino acid change creates a small crack in the p53 protein, making it thermally unstable at physiological temperatures and unable to effectively interact with DNA to activate the transcription of target genes. By selectively binding to the crack, Compound 1 stabilizes the mutant protein structure in the wild-type (WT) conformation, thereby restoring the function of the mutant p53.

[0304] Compound 1 is supplied in 50 mg, 150 mg, and 500 mg tablets. The study drug is provided in appropriately labeled bottles. Patients receive the corresponding study drug supply according to the dosing instructions based on the prescribed dosage. Compound 1 is administered orally in a continuous 21-day dosing cycle. The initial dosing schedule is once daily, but may be adjusted to a more frequent or less frequent schedule as needed during the study period. When administered, all doses of Compound 1 are taken orally at approximately the same time each morning, with food restricted for 2 hours before and 1 hour after administration. Whenever possible, all doses of Compound 1 are taken with approximately 240 mL of water. Patients continue to receive Compound 1 as long as clinical benefit is demonstrated or until disease progression or other criteria for treatment discontinuation are met.

[0305] This study selectively recruited patients with advanced cancer who had been confirmed (historically or prospectively) to have tumors carrying the p53 Y220C mutation prior to signing the primary study informed consent form. Patients had demonstrated solid malignancies with the p53 Y220C mutation, identified by molecular testing performed by a Clinical Laboratory Improvement Amendments (CLIA) certified laboratory. The p53 Y220C mutation originated from the tumor and was not a result of suspected clonal hematopoiesis (CHIP) of undetermined potential.

[0306] CHIP is a common age-related phenomenon in which hematopoietic stem cells or other early blood cell progenitors contribute to the formation of genetically distinct subsets of blood cells. CHIP can explain up to 25% to 40% of TP53 variants detected in saliva and blood during germline testing and may require additional confirmatory testing. Among TP53 mutations, Y220C is one of the most common mutations in CHIP. If the assay does not evaluate CHIP, the liquid assay detects p53 Y220C derived from CHIP rather than tumor.

[0307] p53 Y220C Testing: Blood samples were collected from eligible patients during screening for retrospective germline sequencing to determine the presence of the p53 Y220C mutation using molecular testing. Germline results were not an entry requirement for the study, and all patients were initiated with Compound 1 before results became available. Genetic counseling and additional confirmatory testing using tissue from skin biopsies were provided to patients with positive or indeterminate germline tests. Germline testing is exploratory and provides a basis for understanding whether germline p53Y220C mutations increase patient susceptibility to Compound 1 toxicity due to systemic reactivation of p53. All patients were closely monitored for toxicity during frequent clinical visits, with standard safety assessments. If toxicity was observed, appropriate action was taken, including interruption and / or discontinuation of treatment.

[0308] Acceptable patient identification assays include: 1) solid tissue assays performed in CLIA-certified laboratories, or 2) ctDNA assays performed in CLIA-certified laboratories including a matched normal erythrocyte sedimentation rate (ESR) tannin layer sample. Patients with an identified p53 Y220C mutation in their tumors are eligible for screening. Patients with a p53 Y220C mutation identified by different assays (e.g., confirmed via a p53 Y220C identification form) require a pre-screening step. Patients are identified using the Resolution ctDX Lung™ blood-based test or the FoundationOne® CDx tissue test to prospectively confirm the presence of a p53 Y220C mutation. The Resolution ctDX Lung™ test involves sequencing of plasma and a matched normal ESR tannin layer to determine whether the p53 Y220C mutation originates from a tumor. Based on FoundationOne. ® Tissue-based CTA for CDx testing uses either fresh biopsy or archived tissue. The decision to perform a pre-screening biopsy is based on the patient providing written informed consent for pre-screening and the investigator's assessment that the procedure meets the criteria for care (i.e., low-risk criteria and tumor size sufficient to proceed). Following informed consent for the primary study, a blood sample is collected from the patient during screening for retrospective testing to confirm the presence of the p53 Y220C mutation using the Resolution ctDX Lung™ test. Patients with available archived tumor tissue are sent to the FoundationOne-based CTA. ® CTA for CDx testing is used for retrospective confirmatory testing. If archived tumor tissue is unavailable, a fresh tumor biopsy is required for retrospective confirmatory testing if the biopsy procedure meets the criteria for care (i.e., low-risk criteria and the tumor is of sufficient size for biopsy). If available, the sample used for retrospective confirmatory testing is replaced with tissue biopsy and / or archived tumor tissue collected during pre-screening.

[0309] To explore whether the presence of additional p53 mutations or non-p53 mutations is related to clinical outcomes, pretreatment samples were analyzed using a genome of 324 genes included in a FoundationOne® CDx molecular assay. Additional genomic analyses included loss of heterozygosity (LOH), somatic-germline zygosity (SGZ), microsatellite instability (MSI), and tumor mutational burden (TMB).

[0310] Resolution ctDX Lung TM The test contained a large tumor genome, including p53. In addition to using Resolution ctDX Lung... TMIn addition to the confirmatory p5 Y220C test, this test was also used to explore the potential relationship between other p5 (and non-p53) mutations and the response to treatment with compound 1. Furthermore, blood was collected during compound 1 treatment and analyzed via Resolution ctDX Lung. TM The tests were analyzed to explore whether circulating p53 Y220C levels could serve as surrogate biomarkers for efficacy or disease progression. ctDNA analysis also explored whether the presence of p53 (and non-p53) mutations altered the response to compound 1 treatment.

[0311] Patients with advanced solid tumors harboring the p53 Y220C mutation were recruited for a phase 1 clinical trial administering compound 1. Patients were 12 years of age or older and had locally advanced or metastatic solid tumors harboring the p53 Y220C mutation. Up to 60 patients were recruited in phase 1 to determine the maximum tolerated dose (MTD) and the recommended phase 2 dose (RP2D). The primary objectives of this study were to determine the maximum tolerated dose (MTD), select the recommended phase 2 dose (RP2D), and evaluate the safety and tolerability of compound 1.

[0312] Primary endpoints included the incidence of dose-limiting toxicities (DLT), adverse events (AEs), serious adverse events (SAEs), and changes between baseline and post-baseline laboratory assessments, electrocardiograms (ECGs), vital signs, and physical examinations. Toxicity was graded using the Common Terminology Criteria for Adverse Events (CTCAE) v5.0. Secondary endpoints included characterizing the pharmacokinetics (PK) of compound 1 upon oral administration; describing the concentration of compound 1 upon oral administration; and evaluating the preliminary potency of compound 1 using tumor response criteria. Secondary endpoints were evaluated using plasma PK parameters, including but not limited to: the observed maximum plasma concentration (C0.05). max ), time to observe maximum plasma concentration (T) max The time from time 0 to the final quantifiable concentration or the area under the concentration-time curve (AUC) within a dosing interval. 0-T AUC tau ), observed peak and trough concentrations (C 峰谷 / C tauCompound 1 concentration; and overall response rate (complete response [CR] + partial response [PR]; ORR) and time to response (CR+PR; TTR), duration of response (DoR), disease control rate (CR+PR+stable disease [SD]; DCR), and progression-free survival (PFS) at 18 weeks, as determined by RECIST v1.1, as assessed by blinded independent central review (BICR), or as modified by Prostate Cancer Working Group 3 (PCWG3) RECIST v1.1, as assessed by BICR. A partial response is a response with a reduction in tumor size of at least 30%. TTR is defined as the time from the start of study treatment to the first recorded response (CR+PR), followed by confirmation. DoR is defined as the time from the recording of objective tumor response (CR or PR, whichever occurs first) to the first occurrence of radiographic disease progression or death from any cause. Confirmation of CR or PR is required at ≥4 weeks; however, the date of recording of objective tumor response is based on the date the criteria were met prior to confirmation. If the minimum duration of SD is 6 weeks (±1 week) from the start of treatment, then DCR is defined as the percentage of patients with a complete or partial response or stable disease (CR+PR+SD) at 6, 12, 18, and 24 weeks. PFS is defined as the time from the start of study treatment to the first occurrence of radiological disease progression or death from any cause (whichever comes first).

[0313] Exploratory endpoints include evaluating the effects of compound 1 on multiple pharmacodynamic (PDc) biomarkers of compound 1 activity in blood and / or tumor tissue; exploring the exposure-response relationship between plasma exposure to compound 1 and the safety, PDc biomarkers, and preliminary potency of compound 1; and exploring the PK metabolites of compound 1 in plasma. Exploratory endpoints include circulating tumor DNA (ctDNA) (e.g., Y220C, other p53 mutations, non-p53 mutations), p53 target gene and protein expression (e.g., total p53, p21, macrophage inhibitory cytokine-1 [MIC-1], and multiple biomarkers of immune cell activation) in patient blood, circulating tumor cells (CTCs), and tumor samples collected before and during treatment with compound 1; and the presence or concentration of additional pharmacokinetic metabolites.

[0314] Phase 1 recruitment was limited to patients with advanced solid tumors whose cancers had a p53 Y220C mutation identified by NGS. Phase 1 consisted of an accelerated titration design in the initial dose group, followed by a modified probability toxicity interval (mTPI) design in subsequent dose groups. This approach minimized the number of patients treated at potentially subtherapeutic dose levels. Adolescent patients aged 12 to 17 years and ≥40 kg were eligible to be recruited into the mTPI portion of Phase 1 upon SRC approval. Prior to recruiting adolescent patients, the SRC reviewed safety data and available PK data.

[0315] The dose levels for Phase 1 are summarized in Table 3. The starting dose of Compound 1 is 150 mg once daily (QD). If the first patient is intolerant to 150 mg, the patient will be given the opportunity to reduce the dose to 100 mg upon recovery, provided that the dose modification criteria are met. Due to the presence of methanol as a residual solvent in the pharmaceutical product, the maximum permissible dose of Compound 1 is 5,000 mg daily. The maximum permissible dose is within the daily permissible exposure limits set by the ICH Q3C guidelines for Class 2 solvents such as methanol. Based on an allometric scaling from data from nonclinical studies, pharmacological activity in humans is planned to be observed at a dose level of approximately 570 mg of Compound 1. Therefore, the maximum permissible dose of Compound 1 at 5,000 mg is significantly higher than the planned effective dose of Compound 1. Figure 1 The figure illustrates the design of Phase 1 and Phase 2 studies of compound 1 administration in patients with advanced solid tumors harboring the p53 Y220C mutation (BICR: blinded independent central review; BID: twice daily; CTC: circulating tumor cells; ctDNA: circulating tumor DNA; DCR: disease control rate; DoR: duration of response; MTD: maximum tolerated dose; mTPI: modified probabilistic toxicity interval design; ORR: objective response rate according to RECIST (Responsiveness Evaluation Criteria for Solid Tumors) 1.1; OS: overall survival; PFS: progression-free survival; PK: pharmacokinetics; QD: once daily; RP2D: recommended Phase 2 dose; TTR: time to response). OS was defined as the time from the start of study treatment to death from any cause. For those patients who did not die or were lost to follow-up before the end of the study, OS was checked on the last day of known patient survival or at the data cutoff date (whichever occurred first).

[0316] Table 3 Molecular profiling of tumors in relation to KRAS status and treatment efficacy: Molecular profiling of tumors from stage 1 patients was performed to assess... KRAS The impact of mutation states on the response. KRAS Mutations are defined as single nucleotide variants (SNVs). TP53Y220C / KRAS wild type (WT) power-evaluable analysis sets include those with measurable disease at baseline (without) KRAS Patients with a post-baseline tumor assessment ≥1 within the effective dose range (SNV).

[0317] Sixty-seven patients were treated within the effective dose range (1150 mg QD to 1500 mg BID). Among patients within the effective dose range, the median age was 63 years (range 32–84 years), 61% were female, 76% were white, 67% had an ECOG performance status of 1, and the median number of prior lines of systemic therapy was 3 (range 1–9) (Table 4).

[0318] Table 4 *Including sarcoma, bile duct cancer, esophageal cancer, gastric esophageal cancer, germ cell tumors, rhabdomyosarcoma pleomorphus, small intestinal cancer, and urothelial carcinoma.

[0319] ECOG, Eastern Cooperative Oncology Group; SNV, Single Nucleotide Variant.

[0320] Accelerated Titration: To minimize the number of patients treated at potentially subtherapeutic dose levels, the initial portion of Phase 1 follows an accelerated titration design, whereby each cohort includes a minimum of one and a maximum of two patients. During the accelerated titration portion of Phase 1, at least one patient must have completed Cycle 1 (C1), and safety and DLT (dose-to-weight-leverage) must have been assessed for at least 3 weeks prior to the start of recruitment for the next cohort (including the pre-dosing assessment at C2D1). A second patient is eligible for recruitment into the same cohort if they can begin treatment within 2 weeks of the first patient's treatment in the same cohort. A sentinel approach is used in Cohort 1, whereby the first patient must have completed at least one week of treatment, and a safety assessment must have been reported at C1D8 prior to the start of treatment for the second Cohort 1 patient.

[0321] If a second patient is treated in any dose group, the safety of both patients is fully evaluated before opening a subsequent dose group. The accelerated titration portion continues until grade ≥2 drug-related toxicity is observed in ≥1 patient during the 21-day DLT observation period (C1). Once grade ≥2 drug-related toxicity is observed, the accelerated titration portion ends, and the mTPI portion begins at a dose level no higher than the level at which grade ≥2 toxicity was observed. If the dose levels are the same, data from the accelerated titration portion of the study are included in the dose escalation decisions related to the mTPI design.

[0322] If preliminary efficacy is observed during the accelerated titration portion of the study in the absence of ≥ grade 2 drug-related toxicity during the 21-day DLT observation period (C1), the study transitions to the mTPI portion of the study, which begins with the next highest dose group to collect additional safety, PK, PDc, and preliminary efficacy data in subsequent groups.

[0323] mTPI: mTPI is a rule-based approach that uses Bayesian modeling to define dosing decisions. The Phase 1 mTPI portion guides the selection of dose levels until the MTD is determined based on mTPI rules, the dose escalation portion of the study is stopped, or the maximum permissible dose of Compound 1 is reached. If ≥ Grade 2 drug-related toxicity is observed during the accelerated titration period, the first dose cohort of the mTPI portion uses a dose no higher than that of the highest dose cohort used during the accelerated titration period. Initially, 3 to 4 patients are recruited for each mTPI dose cohort. If a fourth patient can be started within 2 weeks of the third patient starting treatment, that patient is recruited into the cohort. To determine that a dose level is safe and tolerable, at least 3 patients treated at that dose level are required to complete a 21-day DLT period (C1) and a pre-dosing assessment for C2D1. If a fourth patient is treated in any dose cohort, the safety of all 4 patients is fully evaluated before making future dosing decisions.

[0324] During the mTPI phase, pre-defined rules guide the selection of the dose level to be tested, associated with dose decisions based on statistical modeling using all safety data. Dosing decisions (i.e., escalation, descent, or maintenance) are based on observed toxicity outcomes and a predefined set of operational characteristics. Additionally, PK and PDc data are reviewed, when available, to help inform dose selection decisions. At the end of Phase 1, the MTD is estimated as the highest dose among the tested doses with estimated toxicity less than or equal to the target toxicity equivalence interval (e.g., ≤35%). The mTPI design is eligible to be stopped, and the MTD determines whether at least nine patients have been recruited and evaluated in a given dose cohort, and whether the mTPI decision is “maintain.” A maximum of 12 patients are recruited in a single dose cohort. During the mTPI phase, if the current cohort being evaluated is filled and another patient who may be eligible for the study is identified, the potential patient undergoes screening to be recruited into the next lowest dose cohort, provided that recruitment in the lower dose cohort does not exceed 12 patients and any lower dose cohort has shown preliminary evidence of PDc activity and / or preliminary potency.

[0325] Any dose-dependent exposure (DLT) experienced by patients recruited from previously cleared cohorts is not included in subsequent dose escalation decisions or the definition of the mean time to dose (MTD). However, patient data are reviewed and taken into account when assessing safety and tolerability and selecting the RP2D. Dosing may be discontinued prior to establishing the MTD if safety concerns arise, or if clinical and PDc data indicate that the optimal bioactive dose has been reached, or if further increases in exposure are unlikely with further dose increases.

[0326] Serial blood samples were collected from all patients (including adolescents) for PK assessment. Serial blood samples were also collected from all patients for PDc assessment of response to compound 1. Pre-treatment and in-treatment tumor biopsies for PDc assessment were optional. The relationship between plasma compound 1 concentration and pharmacodynamic biomarkers of response was explored. When the MTD was established, this cohort was expanded to up to 12 patients. At least one additional cohort of patients with total daily doses below the MTD and deemed active was expanded to up to 12 patients. These expanded cohorts provided additional information on the relationship between dose, safety / tolerability, and efficacy to inform the selection of RP2D. Data from Phase 1 were reviewed to assess the benefit-risk of each dose level when selecting RP2D. Based on a review of all available safety, PK, PDc, and preliminary efficacy data, the RP2D was determined as the dose leading to the optimal therapeutic window for compound 1. The RP2D may be the same as or below the MTD.

[0327] Intra-patient dose escalation: During Phase 1, intra-patient dose escalation is permitted if the patient tolerates Compound 1 well (i.e., experiences no drug-related grade ≥2 adverse events) and treatment continues for at least 2 cycles. The patient's dose is increased to a level that has been determined to be safe and tolerable. Whenever a patient undergoes a dose escalation, the patient is required to undergo weekly safety assessments at the new dose level during the first two cycles, as outlined in Cycles 1 and 2.

[0328] Phase 1 Dose Elevation: Phase 1 employs an accelerated titration design in the initial dose escalation cohort until grade ≥2 drug-related toxicity occurs. In subsequent dose escalation cohorts, an mTPI design is used, starting at a dose level no higher than the dose level at which grade 2 toxicity occurs. The mTPI design described above is used to determine the MTD. The mTPI design employs a simple β-binomial stratification model, where the decision rule is based on calculating the unit probability mass (UPM) of three intervals corresponding to underdosing, appropriate dosing, and overdosing in terms of toxicity. The three dosing intervals are defined by equivalent intervals reflecting the uncertainty surrounding the true target toxicity and are associated with three distinct dose level decisions. Underdosing intervals correspond to dose escalation, overdosing to dose decrement, and appropriate dosing to maintaining the current dose. After each patient cohort, the mTPI design calculates the UPM for the three dosing intervals, and the interval with the largest UPM defines the corresponding dose discovery decision. The dose escalation portion of the study is terminated when the lowest dose exceeds the MTD or the predetermined maximum sample size is reached. The phase 1 mTPI portion includes up to 60 patients with a target DLT rate of 30%, an equivalent interval of ±5%, and a cohort size of 3 or 4. Due to the rarity of the p53 Y220C mutation, if the mTPI design rules require recruitment of more than 9 patients for a single dose, the mTPI design is eligible to be stopped and an MTD selected.

[0329] Compound 1 concentration data are summarized by dose level and study day using nominal time points. Individual and mean concentrations are presented as plots relative to nominal time by dose level. Pharmacokinetic parameter values ​​are derived using non-compartmental methods via validated pharmacokinetic analysis procedures. All formal analyses use actual time. PK parameters are summarized by dose level. Each PK parameter (e.g., Cp) is listed by dose level. max T max AUC 0-T AUC tau C 峰谷 / C tau Individual values ​​for N (cumulative ratio) and the following (non-model-based) descriptive statistics are provided: N (number of patients with non-missing data), arithmetic mean, standard deviation, arithmetic percentage coefficient of variation (CV) (calculated as 100 × standard deviation / arithmetic mean), median, minimum, maximum, geometric mean, and geometric percentage CV (calculated as: Where s 2 (This is the variance observed on the natural logarithmic scale).

[0330] The operational characteristics associated with the mTPI design were evaluated at different maximum sample volumes when (i) all dose levels had a true toxicity rate equal to 10% and (ii) the dose levels had a monotonically increasing true toxicity rate. The operational characteristics of the mTPI design were evaluated based on the mTPI portion of a study with five dose levels.

[0331] The most frequent treatment-related adverse events (TRAEs) (>20%) were nausea, vomiting, and increased serum creatinine. Administration of Compound 1 with food resulted in improved gastrointestinal toxicity (nausea, vomiting, and diarrhea), with a similar safety profile observed within the effective range. A 3% drug discontinuation rate due to TRAEs was observed. Table 5 summarizes the incidence of TRAEs in patients receiving Compound 1 at doses ranging from 1150 mg QD to 1500 mg BID.

[0332] Table 5 † This includes five additional Grade 3 treatment-related adverse events: decreased neutrophil count, acute kidney injury, pancreatitis, pneumonia, and erythematous rash. Note that patients may experience multiple Grade 3 events.

[0333] ‡ This includes a patient with grade 4 immune thrombocytopenic purpura.

[0334] Data cutoff: September 5, 2023. ALT, alanine aminotransferase; AST, aspartate aminotransferase; BID, twice daily; CTCAE, Common Terminology Standard for Adverse Events; QD, once daily; TRAE, treatment-related adverse events.

[0335] Compound 1 exhibited dose-proportioning and linear p-p-values ​​from 150 mg to 2500 mg QD at steady state. Figure 2 ). Figure 2 Day 15 (steady-state) exposure to compound 1 was plotted. The median half-life of compound 1 at steady state (day 15) at each dose was 19 hours (AUC, area under the curve; SD, standard deviation).

[0336] Clinical efficacy was achieved in a large number of pre-treatment patients across multiple tumor types. Within the safety cohort (N=67), 51 patients had measurable disease at baseline and post-baseline tumor assessment. Of those patients: 38 were included in the TP53 Y220C / KRAS WT efficacy evaluable cohort, and 13 had tumors carrying KRAS mutations. Tumor shrinkage was observed across multiple tumor types in the patients with measurable disease included in the efficacy evaluable cohort (N=51), including patients whose tumors were KRAS wild-type. Figure 3 ).

[0337] Figure 3 This includes patients with measurable disease at baseline and a post-baseline assessment ≥1. One patient without tumor measurement at the first scan was not represented. “Other” tumor types include sarcoma, esophageal cancer, germ cell tumors, rhabdomyosarcoma pleomorphus, and urothelial carcinoma. BID, twice daily; PD, progressive disease; PR, partial response; QD, once daily; SD, stable disease; WT, wild-type.

[0338] In patients with TP53 Y220C mutation and KRAS WT disease, a total of 13 confirmed partial responses (PRs) were observed across multiple tumor types, including ovarian cancer, breast cancer, small cell lung cancer, and endometrial cancer (overall response rate [ORR] = 34%) (Table 6).

[0339] Table 6 * All partial responses were confirmed. Only tumor types with at least one responder were included in the overall response analysis across tumor types. KRAS WT potency was evaluable: all treated patients had measurable disease at baseline, KRAS WT was defined as no KRAS SNV, and post-baseline assessment ≥1. BID, twice daily; ORR, objective response rate; PD, progressive disease; PR, partial response; QD, once daily; SD, stable disease; SNV, single nucleotide variant; WT, wild-type.

[0340] With 2000 mg QD, the ORR was 38% in patients with TP53 Y220C and KRAS WT tumors.

[0341] Clinical activity was observed in patients with KRAS mutations, but no confirmed response was reported. Clinical activity or clinical efficacy is activity identified across various trial endpoints or tumor response criteria, including ORR, time to response (TTR), duration of response (DoR), disease control rate (DCR) at 6, 12, 18, and 24 weeks, and progression-free survival (PFS) and overall survival (OS) based on BICR.

[0342] Among all respondents, the median response time and median response duration were 1.5 months and 7 months, respectively. Figure 4 ). Figure 4 A summary of TP53 Y220C / KRAS WT efficacy can evaluate target lesion reduction across tumor types in a population (1150 mg QD to 1500 mg BID), and includes patients with measurable disease at baseline. KRASWT and all patients with a post-baseline assessment ≥1 (n=38). "Other" tumor types included 1 patient with a germ cell tumor, 2 patients with sarcomas, 1 patient with urothelial carcinoma, and 1 patient with esophageal cancer.

[0343] Treatment with compound 1 within the effective dose range and is TP53 Y220C / KRAS Of the 51 patients with WT, 40 had available ctDNA TP53 Y220C variant allele frequencies (VAF) at baseline. Figure 5 ). Figure 5 It was shown in week 3 TP53 Percentage change in Y220C VAF relative to baseline. CFBL, change relative to baseline; ctDNA, circulating tumor DNA; NGS, next-generation sequencing; PR, partial response; QD, once daily; VAF, variant allele frequency. 80% (n=32) and 42.5% (n=17) of patients showed changes in TP53 Y220C VAF relative to baseline of ≥50% and ≥95%, respectively. 92.5% (n=37) of patients had a reduction in TP53 Y220C VAF, indicating target activity. Furthermore, all patients who experienced PR had a reduction in TP53 Y220C VAF.

[0344] Case Studies A 51-year-old woman presented with metastatic triple-negative breast cancer (TNBC). She had previously undergone neoadjuvant therapy (carboplatin, paclitaxel, and pembrolizumab, followed by cyclophosphamide, doxorubicin, and pembrolizumab), bilateral mastectomy, followed by pembrolizumab maintenance, radiation therapy, and breast reconstruction, and treatment with pegylated liposomal doxorubicin for disease recurrence. Progressive disease was observed in the axilla, with extensive skin lesions on adjacent breasts and arms, limiting mobility. TP53 Y220C was detected by next-generation sequencing. A QD of 2000 mg of compound 1 was initiated. Rapid, visible reduction in arm swelling and improvement in arm and finger mobility were observed within the first week. Partial response (PR) was observed at week 6 (41% reduction in axillary lesions) and was confirmed and sustained at week 12. Figure 6 Images of patients with triple-negative breast cancer at baseline and after the third cycle of treatment with compound 1 are provided.

[0345] in conclusion Compound 1 demonstrated a favorable safety profile within the effective dose range, with improved gastrointestinal adverse events when taken with food. Single-dose clinical efficacy was achieved in a large number of pre-treated patients across multiple tumor types.

[0346] Example 3: Activity of compound 1 in ovarian cancer In a subset of 22 patients with ovarian cancer treated in the Phase 1 clinical trial detailed in Example 3, preliminary efficacy (ORR) and serum CA-125 response (defined as a >50% reduction at two separate time points, 4 weeks apart) of RECIST v1.1 as assessed by the investigator were analyzed. Safety across tumor types was also evaluated within the effective dose range. This subgroup analysis investigated the efficacy of compound 1 in patients with advanced ovarian cancer treated within the effective dose range (1150 mg daily to 1500 mg twice daily).

[0347] The median age of patients with ovarian cancer (N=22) was 66 years (range 49–81 years). At baseline, 20 patients had high-grade serous ovarian cancer and 2 had endometrioid carcinoma; 2 had BRCA2 mutations and 20 had no BRCA1 / 2 mutations; 6 were HRD-positive, 11 were HRD-negative, and 5 had unknown HRD status. All were KRAS WT. The median number of prior lines of systemic therapy was 4 (range 1–9). At the start of the study, 19 patients were platinum-resistant, 2 were platinum-refractory, and 1 was platinum-sensitive. Other patient demographics and disease characteristics are summarized in Table 7. * Patients whose disease progressed within 6 months of platinum-based chemotherapy. ** Patients whose disease progressed during platinum-based therapy or within 4 weeks after the last dose. † Defined as none. KRAS Single nucleotide variants. ECOG, Eastern Cooperative Oncology Group; HGSOC, High-grade serous ovarian cancer; HRD, Homologous recombination defect.

[0348] The results showed that among the 15 patients with measurable disease at baseline and a post-baseline tumor assessment ≥1 (evaluable efficacy group), 7 patients achieved a confirmed partial response (PR), 7 had stable disease (SD), and 1 had progressive disease. Figure 7 This is a graph summarizing the best percentage change in the total number of tumor target lesions among 15 patients in a subgroup of ovarian cysts with measurable disease. Figure 7 In the text, * indicates a patient who experienced an initial, unconfirmed, and subsequently unconfirmed partial response. NE, non-evaluable; ORR, overall response rate; PD, progressive disease; PR, partial response; SD, stable disease.

[0349] The median duration of response was 7 months. Of the 15 patients with measurable serum CA-125 at baseline, 6 had a CA-125 response. Of these patients, 5 achieved radiological partial response (PR), and one had stable disease (SD). Figure 8 This is a graph summarizing the best percentage change in CA-125 levels in 15 patients in a subgroup of ovarian cysts with measurable CA-125 at baseline. * CA-125 response was defined as a 50% reduction in CA-125 relative to baseline at two separate time points 4 weeks apart.

[0350] Figure 9 This is a graph summarizing response time and treatment duration for patients with a post-baseline tumor assessment ≥1 in the ovarian cyst subgroup with measurable disease at baseline. The median duration of response was 7 months.

[0351] In the overall cohort of 67 patients assessed within the effective dose range (≥1150 mg QD) (including a subset of ovarian cancer patients), the majority of treatment-related adverse events (TRAEs) were grade 1 and 2, and the majority of TRAEs were nausea (51%), vomiting (43%), and increased serum creatinine (27%). The frequency and severity of TRAEs were similar in the ovarian cancer cohort compared to the overall cohort. Administration of compound 1 with food resulted in improvement in nausea and vomiting.

[0352] Example 4: Pharmacokinetics of compound 1 in fasting and eating subjects.

[0353] The pharmacokinetics of compound 1 were analyzed in healthy subjects who were administered it while fasting. A subject was considered fasting if they had fasted overnight for at least 10 hours and were permitted to drink water freely except for one hour before and after administration. For fasting subjects, compound 1 was administered with at least 8 ounces (240 mL) of water. The pharmacokinetics of compound 1 were also analyzed in healthy subjects who were administered it while eating. A subject was considered to have eaten if they consumed a high-fat meal 30 minutes before administration and completed the meal at least 5 minutes before administration. For both fasting and eating subjects, food was not permitted for at least 4 hours after administration, and water was permitted starting 1 hour after administration.

[0354] The participants were healthy, non-smoking male and female Caucasian / non-Asian or Japanese participants, aged 18 to 55 years, with a BMI of 18.5 kg / m². 3 and 30.0 kg / m 3 between.

[0355] Part 1 Study Design (1000 mg Compound 1) In Part 1 of the study, analyses were performed to compare 1000 mg of Compound 1 in fed and fasted states. The 1000 mg dose of Compound 1 consisted of two 500 mg tablets administered as a single oral dose. The analyses in Part 1 were performed at two separate time points in two different protocols (1A and 1B).

[0356] In Protocol 1A, Compound 1 was analyzed in 3 healthy subjects (n=3) who were given 1000 mg of Compound 1 while on an empty stomach during time period 1. After a clearance period of 7–9 days, these subjects were then given 1000 mg of Compound 1 while on an empty stomach.

[0357] In Protocol 1B, compound 1 was analyzed in healthy subjects (n=3) who were administered 1000 mg of compound 1 while fasting during time period 1. After a clearance period of 7–9 days, these subjects were then administered 1000 mg of compound 1 while eating.

[0358] Part 2 Study Design (2000 mg Compound 1) In Part 2 of the study, analyses were performed to compare 2000 mg of Compound 1 in fed and fasted states, and also to compare the ethnicity of Caucasian / non-Asian subjects and Japanese subjects. The 2000 mg dose of Compound 1 consisted of four 500 mg tablets administered as a single oral dose. Participants were classified as Japanese if (i) they were born in Japan, (ii) their parents and grandparents were of Japanese ethnicity and they were born in Japan, (iii) they had lived outside Japan for less than 10 years, and (iv) their lifestyle had not undergone significant changes since leaving Japan.

[0359] For Caucasian / non-Asian subjects (n=20), analyses were performed at two separate time points across two distinct protocols (2A and 2B). In Protocol 2A, during time period 1, Compound 1 was analyzed in subjects (n=10) who received 2000 mg of Compound 1 while on an empty stomach. Following a 7–9 day clearance period, these subjects received 2000 mg of Compound 1 again during time period 2, while on an empty stomach. Separately, in Protocol 2B, Compound 1 was analyzed in subjects (n=10) who received 2000 mg of Compound 1 while on an empty stomach during time period 1. Following a 7–9 day clearance period, these subjects received 2000 mg of Compound 1 again during time period 2, while on an empty stomach.

[0360] The pharmacokinetics of compound 1 were analyzed in Japanese subjects (n=6) who received 2000 mg of compound 1 while fasting.

[0361] Pharmacokinetic (PK) Analysis - Blood Samples Blood was collected for analysis of the research drug (compound 1) and metabolite (M1).

[0362] In Part 1 of the study, feeding / fasting conditions (linear and semi-log scales) were used ( Figures 10-11 ), and in Part 2 of the study, by race and eating / fasting conditions ( Figures 12-13 The arithmetic mean (±SD) plasma concentration-time plot of compound 1 was grouped.

[0363] Following administration of a single oral dose of compound 1 in Parts 1 and 2, plasma concentrations of compound 1 rapidly reached a peak and then declined in a generally biexponential manner. For both the 1000 mg and 2000 mg doses, Caucasian / non-Asian participants had higher mean concentrations of compound 1 throughout the entire PK profile under fed conditions compared to fasting conditions. Figures 10-13 ).

[0364] Following administration of a 2000 mg dose under fasting conditions, Japanese participants had higher mean plasma concentrations of compound 1 compared to Caucasian / non-Asian participants. Figures 12-13 ).

[0365] Determine various pharmacokinetic (PK) parameters for each subject. (Tables 11-14 and...) Figures 14-19 For example, the terminal elimination rate constant (λ) z ); From exceeding t last AUC obtained by extrapolation 0-inf Percentage (AUC) %extrap The area under the concentration-time curve (AUC) from before administration (time 0) to 24 hours later. 0-24 The area under the concentration-time curve (AUC) from before administration (time 0) to 96 hours later. 0-96 The area under the concentration-time curve (AUC) extrapolated from before administration (time 0) to infinity. 0-inf From before administration (time 0) to t last Area under the concentration-time curve (AUC) 0-last ); Apparent total body clearance (CL / F); Observed maximum plasma concentration (C max ); Apparent terminal elimination half-life (t½); Time before first measurable (non-zero) concentration (t lag The final time for quantifying the concentration (t) last ); corresponding to C max Time of occurrence (T) max ); and apparent distribution volume (V z / F).

[0366] In Part 1 of the study, the statistical significance of plasma PK parameters of Compound 1 was evaluated based on feeding / fasting conditions (Table 8) and in Part 2 of the study, based on race and feeding / fasting conditions (Table 9). For Part 1, the geometric mean (gCV%)C after administration of a single 1000 mg oral dose of Compound 1 max The values ​​under fasting and fed conditions were 8690 ng / mL (19.0%) and 10600 ng / mL (18.2%), respectively. Median T max The values ​​were 2.27 hours (range: 1.54 hours to 2.51 hours) under fasting conditions and 3.25 hours (range: 1.51 hours to 5.01 hours) under feeding conditions. Geometric mean (gCV%) AUC 0-last The values ​​under fasting and feeding conditions were 147,000 h·ng / mL (23.4%) and 177,000 h·ng / mL (32.6%), respectively.

[0367] Geometric mean (gCV%) AUC 0-inf The values ​​under fasting and fed conditions were 150,000 h·ng / mL (23.2%) and 180,000 h·ng / mL (33.4%), respectively. The percentages obtained by extrapolation for both fasting and fed conditions (AUC) are... %extrap Less than 5%. Low variability of the major plasma PK parameters was observed in both groups (less than 23.4% for fasting conditions and 33.4% for fed conditions). Under fasting conditions, plasma CL / F and V z / F is slightly higher.

[0368] For Part 2, for Caucasian / non-Asian participants, the geometric mean (gCV%)C following administration of a single 2000 mg oral dose of Compound 1 was... max The values ​​were 10300 ng / mL (52.2%) under fasting conditions and 19000 ng / mL (20.7%) under fed conditions. For Caucasian / non-Asian participants, the median T... max The values ​​were 2.52 hours (range: 1.03 hours to 5.94 hours) under fasting conditions and 4.03 hours (range: 2.11 hours to 8.08 hours) under fed conditions. For Caucasian / non-Asian participants, the geometric mean (gCV%) AUC was... 0-lastThe values ​​under fasting and feeding conditions were 254,000 h·ng / mL (53.1%) and 448,000 h·ng / mL (26.4%), respectively.

[0369] For Caucasian / non-Asian participants, the geometric mean (gCV%) AUC 0-inf The values ​​under fasting and fed conditions were 259,000 h·ng / mL (55.5%) and 479,000 h·ng / mL (29.5%), respectively. The percentages (AUC) obtained by extrapolation for both fasting and fed conditions are... %extrap Less than 17%. Low to moderate variability in the major plasma PK parameters was observed in both groups (less than 55.5% and 29.5% for fasting and fed conditions, respectively). Plasma CL / F and Vz / F were slightly higher under fasting conditions.

[0370] For Caucasian / non-Asian participants, the maximum exposure to compound 1 (C1) under the fed condition (high-fat diet) compared to the fasting condition was [missing information]. max Both total exposure (AUC) and systemic exposure were numerically higher under feeding conditions with prolonged T... max A slight delay was observed in the absorption (the delay was approximately 1.00 hours for part 1 and 1.50 hours for part 2).

[0371] For Japanese participants under fasting conditions, compared with Caucasian / non-Asian participants under fasting conditions, the peak exposure (C1) of compound 1 in Japanese participants was significantly lower. max The values ​​for total exposure (AUC) and overall exposure (AUC) were numerically higher. No formal statistical comparisons were performed during the study.

[0372] Analysis of the food effect Statistical analyses of the plasma PK parameters of compound 1 were performed to evaluate the food effects in Part 1 (Table 10) and Part 2 (Table 11). Analysis of variance models were used to analyze the log-transformed PK parameters, with treatment (eating / fasting conditions) and time period as fixed effects, and participants as random effects.

[0373] Table 10 Table 11 Still on the plasma PK parameters (AUC) of compound 1 0-inf AUC 0-last and C max The trapezoidal diagram is based on the feeding / fasting conditions ( Figures 14-16 And according to race and eating / fasting conditions ( Figures 17-19To evaluate the data.

[0374] Following a single oral dose administered under fasting conditions, compound 1 was rapidly absorbed, with a median T0. max The ranges were from 1.54 hours to 2.51 hours (Part 1) and from 1.03 hours to 5.94 hours (Part 2). The plasma concentration of compound 1 decreased in a bi-exponential manner.

[0375] For Part 1, for AUC 0-inf AUC 0-last and C max The geometric mean ratios of the feeding / fasting conditions were 120.25% (90% CI: 100.89, 143.33), 120.21% (90% CI: 101.26, 142.69), and 121.75% (90% CI: 105.55, 140.44), respectively (Table 10). For Part 2, for AUC 0-inf AUC 0-last and C max The geometric mean ratios of the feeding / fasting conditions were 178.74% (90% CI: 150.88, 211.74), 172.94% (90% CI: 148.55, 201.33), and 183.82% (90% CI: 153.27, 220.46) (Table 11).

[0376] For Part 1, compared to a fasting state, administration of a single dose of 1000 mg of Compound 1 with food (high-fat diet) resulted in a peak exposure (C0.05) of Compound 1. max This increased by approximately 22%, and also increased the total body exposure (AUC) at AUC. 0-inf and AUC 0-last Both aspects increased by approximately 20%. A prolonged T was also observed under dietary conditions (high-fat diet). max (3.25 hours versus 2.27 hours).

[0377] For Part 2, compared to the fasting state, administration of a single dose of 2000 mg of Compound 1 with food (high-fat diet) resulted in a higher peak exposure (C0.05) of Compound 1. max This increased by approximately 84% and increased the total body exposure (AUC) to AUC. 0-inf and AUC 0-last The percentages increased by 79% and 73% respectively. A prolonged T was also observed under dietary conditions (high-fat diet). max (4.03 hours vs. 2.52 hours).

[0378] Analysis of racial effects In Part 2 of the study, between Caucasian / non-Asian participants and Japanese participants, after administration of a single 2000 mg dose of Compound 1 under fasting conditions, peak exposure (C0.05) of Compound 1 was found to be lower in Japanese participants compared to Caucasian / non-Asian participants. max The values ​​for whole-body exposure (AUC) were higher.

[0379] Following administration of a single 2000 mg dose of compound 1 under fasting conditions, the peak exposure (geometric mean) of compound 1 in Caucasian / non-Asian participants was C1. max It is 10300 ng / mL, and the peak exposure (geometric mean) of compound 1 to Japanese participants was C. max The median concentrations were 14700 ng / mL, occurring at 2.52 hours and 3.94 hours, respectively. For Caucasian / non-Asian participants and Japanese participants, the geometric mean AUC was... 0-inf The values ​​were 259,000 h·ng / mL and 338,000 h·ng / mL, respectively, and the geometric mean AUC was... 0-last The values ​​are 254,000 h·ng / mL and 326,000 h·ng / mL, respectively.

[0380] Furthermore, a slight delay in absorption was observed in Japanese participants compared to Caucasian / non-Asian participants (T0). max The duration of illness was prolonged (3.94 hours vs. 2.52 hours). Compared to Caucasian / non-Asian participants, Japanese participants had higher plasma CL / F and V levels. z / F is numerically low.

[0381] Pharmacokinetic (PK) analysis - urine sample Urine was also collected for analysis of the study drug. Participants were asked to urinate at the end of each collection period to ensure complete urine collection at each interval. The total volume of urine excreted at each interval was collected and recorded, and a small volume was stored for analysis of the study drug concentration.

[0382] Determine various pharmacokinetic (PK) parameters for each subject. Figures 20-22 For example, the amount of drug excreted unchanged in urine (Ae), renal clearance (CL) R ) and the fraction excreted in urine (fe).

[0383] The arithmetic mean (±SD) of compound 1 urinary cumulative excretion fraction (%) was assessed relative to the midpoint of the collection interval, grouped as follows: feeding / fasting conditions in Part 1 of the study ( Figure 20 ), the feeding / fasting conditions of Caucasian / non-Asian participants in Part 2 of the study ( Figure 21 ), and the fasting conditions of Japanese participants in Part 2 of the study ( Figure 22 ).

[0384] Following administration of a single dose of compound 1, less than 4% of the dose was excreted in the urine as unchanged compound 1 in all participants (Caucasian / non-Asian and Japanese participants) under both fasting and feeding conditions over a 96-hour interval.

[0385] Following administration of a single 1000 mg dose of compound 1, 2.53% and 3.29% of the total dose administered were excreted in the urine as unchanged compound 1 over a 96-hour interval, under both fasting and fed conditions. Following administration of a single 2000 mg dose of compound 1, 1.90% and 3.66% of the total dose administered were excreted in the urine as compound 1 over a 96-hour interval, under both fasting and fed conditions. Furthermore, for Japanese participants, 2.28% of the total dose administered was excreted in the urine as unchanged compound 1 over a 96-hour interval under fasting conditions.

[0386] For Part 1, renal clearance (CL) R The geometric mean of CLR was 0.172 L / h under fasting and 0.186 L / h under fed conditions. In Part 2, for Caucasian / non-Asian participants, the geometric mean of CLR was 0.152 L / h under fasting and 0.164 L / h under fed conditions. On the other hand, for Japanese participants, the geometric mean of CLR was... R The flow rate is 0.140 L / h under fasting conditions.

[0387] Pharmacokinetic parameters conclusions Following a single oral dose administered under fasting conditions, compound 1 was rapidly absorbed, with a median T0. max The ranges were from 1.54 hours to 2.51 hours (Part 1) and from 1.03 hours to 5.94 hours (Part 2). The plasma concentration of compound 1 decreased in a generally biexponential manner.

[0388] For Caucasian / non-Asian participants, compared with a fasting state, administration of a single dose of 1000 mg of compound 1 with food (high-fat diet) resulted in a higher peak exposure (C0.05) of compound 1. max This increased by approximately 22%, and also increased the total body exposure (AUC) at AUC. 0-inf and AUC 0-last Both aspects increased by approximately 20%. A prolonged T was also observed under dietary conditions (high-fat diet). max(3.25 hours vs. 2.27 hours). Compared to a fasting state, administration of a single 2000 mg dose of compound 1 with food (high-fat diet) increased peak exposure of compound 1 by approximately 84% and increased systemic exposure to AUC. 0-inf and AUC 0-last The increases were 79% and 73% respectively. Furthermore, a prolonged T was also observed under dietary conditions (high-fat diet). max (4.03 hours vs. 2.52 hours).

[0389] Following administration of a single 2000 mg dose of compound 1 under fasting conditions, the peak exposure (geometric mean) of compound 1 in Caucasian / non-Asian participants was C1. max It is 10300 ng / mL, and the peak exposure (geometric mean) of compound 1 to Japanese participants was C. max The median concentrations were 14700 ng / mL, occurring at 2.52 hours and 3.94 hours, respectively. For Caucasian / non-Asian participants and Japanese participants, the geometric mean AUC was... 0-inf The values ​​were 259,000 h·ng / mL and 338,000 h·ng / mL, respectively, and the geometric mean AUC was... 0-last The values ​​are 254,000 h·ng / mL and 326,000 h·ng / mL, respectively.

[0390] Following administration of a single dose of the compound, under both fasting and eating conditions, less than 4% of the dose was excreted in the urine as unchanged compound 1 over a 96-hour interval.

[0391] Example 5: Effects of food on the pharmacokinetics and safety profile of compound 1 in healthy subjects and patients with solid tumors.

[0392] Physiology-based pharmacokinetic (PBPK) modeling and pharmacokinetic (PK) studies were conducted in healthy volunteers and patients with solid tumors under fasting and feeding conditions to assess the potential impact of food on the PK and safety of compound 1.

[0393] Methods and Materials: A PBPK model was developed using in vitro and clinical PK data to predict the effect of a standard diet on compound 1 Pk in healthy volunteers and patients with solid tumors. The model was validated in studies of patients with solid tumors carrying the TP53 Y220C mutation using PK data obtained after single and repeated doses of compound 1 (150 mg–2500 mg); most predictions fell within 1.5-fold of the observed values. A healthy volunteer study was also conducted in healthy volunteers (n=22) receiving a single 2000 mg dose of compound 1 with or without a high-fat diet, with PK samples collected within 96 h post-dose. Patients with advanced solid tumors carrying the TP53 Y220 mutation were also evaluated in studies starting compound 1 (2000 mg, PO, once daily) in a fasting state (n=13) or a feeding state (n=12; ≥100 kcal, 3 g fat), with PK samples collected on day 1 within 24 h post-dose and at steady state.

[0394] Data and Results: Using a PBPK model, 10 virtual trials were generated in 10 healthy volunteers (50% / 50% male / female; age 20–50 years) and 10 virtual trials were generated in 10 patients with solid tumors (50% / 50% male / female; age 30–75 years), who received a 2000 mg dose of compound 1 in either a fasting or fed state. The area under the curve (AUC) predicting food intake in healthy volunteers was calculated from 0 to infinity. 0-inf Increase by 45% and make the maximum concentration (C) max The AUC increased by 28%. For patients with solid tumors, predicting food intake increased the AUC by 28%. 0-inf Increase by 47% and make C max An increase of 35%. Exposure predictions for multiple oral doses showed similar results.

[0395] In the study of healthy volunteers, a single dose of compound 1 (2000 mg) was administered to healthy volunteers (PO) with or without a high-fat diet. The AUC in the fed state was [data missing]. 0-inf and C max They were 79% and 84% higher, respectively.

[0396] In studies of patients with advanced solid tumors carrying the TP53 Y220 mutation, the AUC (AUC) of patients with solid tumors who received 2000 mg of compound 1 at PO after eating was significantly higher from 0–24 h compared to those who had not eaten. 0-24 ) and C max The increases were 40% and 42% higher (at steady state), respectively (Table 12).

[0397] In studies of patients with advanced solid tumors carrying the TP53 Y220 mutation, the rate of treatment-related adverse events (TRAEs) after administration of 2000 mg of compound 1 was similar in patients who ate versus those who did not eat; gastrointestinal (GI) TRAEs were less frequent in patients who ate (Table 12).

[0398] Table 12 Example 6: Analysis of a subgroup of patients with advanced breast cancer Patients with locally advanced or metastatic breast cancer carrying the TP53 Y220C mutation were eligible to receive compound 1 orally on a continuous schedule within the effective dose range (1150 mg once daily [QD] to 1500 mg twice daily). Safety and preliminary potency were assessed (according to RECIST v1.1 as assessed by the investigator). Local next-generation sequencing was used to determine TP53 Y220C, BRCA , PIK3CA and KRAS Tumor mutation status.

[0399] Nine breast cancer patients (HR+ / HER2- n=3; HR+ / HER2+ n=1; HR- / HER2+ n=1; TNBC n=3; unknown n=1) received compound 1 in a phase 1 trial. The median age was 53 years (range 32–65 years); ECOG performance scores were 0 (n=3) or 1 (n=6). Two patients had somatic BRCA2 mutations, no patients had confirmed BRCA1 mutations (two patients had unknown BRCA1 status; one patient had unknown BRCA2 status), two patients had PIK3CA mutations, and all patients were KRAS WT. The median number of prior lines of systemic therapy was 4 (range: 2–9). Of the eight patients with measurable disease at baseline, three achieved a confirmed partial response (PR; overall response rate 37.5%), four had stable disease (SD), and one had progressive disease (PD) as the best objective response. Reduction in target lesions was reported in all patients, with the largest changes in tumor volume relative to baseline ranging from -2.4% to -55.2%. One patient (51 years old) with TNBC achieved PR (41% reduction in axillary lesions) at 6 weeks, confirmed at 12 weeks and ongoing; improvement in symptoms (arm swelling, arm and finger mobility) was observed in the first week of treatment. In the entire Phase 1 cohort (N=67 patients with solid tumors), the most frequent treatment-related adverse events (TRAEs) were nausea (51%), vomiting (43%), and increased serum creatinine (27%), most of which were grade 1 / 2. In the overall cohort, administration of compound 1 with food resulted in improvement in gastrointestinal (GI) adverse events, including nausea and vomiting.

[0400] The following non-limiting embodiments provide illustrative examples of the invention, but do not limit the scope of the invention.

[0401] Implementation Scheme 1. A method for treating a condition in a subject with a corresponding need, the method comprising administering to the subject a therapeutically effective amount of a small molecule compound that binds to a p53 mutant protein, wherein the mutation status of the KRAS gene in the subject has been previously determined.

[0402] Implementation Scheme 2. The method according to Implementation Scheme 1, wherein the mutation status of the KRAS gene in the subject has been previously determined based on assays performed on biological samples of the subject.

[0403] Implementation Scheme 3. The method according to Implementation Scheme 1 or 2, wherein it has been previously determined that the subject has a mutation in the TP53 gene encoding the p53 mutant in the subject.

[0404] Implementation Scheme 4. A method for treating a condition in a subject with a corresponding need, the method comprising: a) determining the mutation status of the KRAS gene in the subject based on the results of a measurement performed on a biological sample of the subject; and b) administering to the subject a therapeutically effective amount of a compound binding to a p53 mutant protein, based at least in part on the mutation status of the KRAS gene in the subject.

[0405] Implementation Scheme 5. The method according to Implementation Scheme 4, wherein the method further comprises determining, based on the results of the assay, that the subject carries a mutation in the TP53 gene encoding the p53 mutant protein, wherein administering a therapeutically effective amount of the compound binding the p53 mutant protein to the subject is based at least in part on determining that the subject carries a mutation in the TP53 gene encoding the p53 mutant protein.

[0406] Implementation Scheme 6. The method according to any one of Implementation Schemes 2, 4 and 5 further includes measuring the biological sample of the subject.

[0407] Implementation Scheme 7. The method according to Implementation Scheme 5 further includes measuring the biological sample of the subject, wherein measuring the biological sample of the subject further includes detecting a mutation in the TP53 gene encoding the p53 mutant in the subject.

[0408] Implementation Scheme 7A. The method according to any one of Implementation Schemes 1-7, wherein the gene is the KRAS gene.

[0409] Implementation Scheme 8. The method according to Implementation Scheme 7A, wherein the mutation status of the KRAS gene in the subject includes the absence of missense mutations at any one of codons 12, 13, 59, 61, 117 and 146.

[0410] Implementation Scheme 9. The method according to Implementation Scheme 7A or 8, wherein the mutational state of the KRAS gene in the subject includes the absence of a mutation at codon 12 that encodes an amino acid other than glycine.

[0411] Implementation Scheme 10. The method according to Implementation Scheme 7A, 8 or 9, wherein the mutational state of the KRAS gene in the subject includes the absence of a mutation at codon 13 that encodes an amino acid other than glycine.

[0412] Implementation Scheme 11. The method according to any one of Implementation Schemes 7A and 9-10, wherein the mutational state of the KRAS gene in the subject includes the absence of a mutation at codon 59 that encodes an amino acid other than alanine.

[0413] Implementation Scheme 12. The method according to any one of Implementation Schemes 7A and 9-11, wherein the mutational state of the KRAS gene in the subject includes the absence of a mutation at codon 61 that encodes an amino acid other than glutamine.

[0414] Implementation Scheme 13. The method according to any one of Implementation Schemes 7A and 9-12, wherein the mutational state of the KRAS gene in the subject includes the absence of a mutation at codon 117 that encodes an amino acid other than lysine.

[0415] Implementation Scheme 14. The method according to any one of Implementation Schemes 7A and 9-13, wherein the mutational state of the KRAS gene in the subject includes the absence of a mutation at codon 146 that encodes an amino acid other than alanine.

[0416] Implementation Scheme 15. The method according to any one of Implementation Schemes 7A and 9-14, wherein the mutation status of the KRAS gene in the subject is the KRAS gene encoding the wild-type K-ras protein.

[0417] Implementation Scheme 16. The method according to Implementation Scheme 15, wherein the wild-type K-Ras protein has a sequence according to SEQ ID NO: 1 or SEQ ID NO: 2.

[0418] Implementation Scheme 17. The method according to any one of Implementation Schemes 7A and 9-16, wherein the mutation status of the KRAS gene in the subject is the copy number variation status of the KRAS gene.

[0419] Implementation Scheme 18. The method according to Implementation Scheme 17, wherein the mutation status of the KRAS gene in the subject is the absence of KRAS copy number variation.

[0420] Implementation Scheme A1. The method according to any one of Implementation Schemes 1-7, wherein the gene is BRCA1.

[0421] Implementation Scheme A2. The method according to Implementation Scheme A1, wherein the mutation status of the BRCA1 gene in the subject is the BRCA1 gene encoding the wild-type breast cancer type 2 susceptibility protein.

[0422] Implementation Scheme A3. The method according to Implementation Scheme A1 or A2, wherein the mutation status of the BRCA1 gene in the subject is a BRCA1 gene that does not encode oncogenic mutations.

[0423] Implementation Scheme A4. The method according to any one of Implementation Schemes A1-A3, wherein the mutation state of the BRCA1 gene in the subject is a state in which there is no copy number variation of the BRCA1 gene.

[0424] Implementation Scheme A5. The method according to any one of Implementation Schemes 1-7, wherein the gene is BRCA2.

[0425] Implementation Scheme A6. The method according to Implementation Scheme A5, wherein the mutation status of the BRCA2 gene in the subject is the BRCA2 gene encoding the wild-type breast cancer type 2 susceptibility protein.

[0426] Implementation Scheme A7. The method according to Implementation Scheme A5 or A6, wherein the mutation status of the BRCA2 gene in the subject is a BRCA2 gene that does not encode oncogenic mutations.

[0427] Implementation scheme A8. The method according to any one of implementation schemes A5-A7, wherein the mutation state of the BRCA2 gene in the subject is a state in which there is no copy number variation of the BRCA2 gene.

[0428] Implementation Scheme A9. The method according to any one of Implementation Schemes 1-7, wherein the gene is PIK3CA.

[0429] Implementation Scheme A10. The method according to Implementation Scheme A9, wherein the mutation status of the PIK3CA gene in the subject is a PIK3CA gene that does not encode oncogenic mutations.

[0430] Implementation Scheme A11. The method according to Implementation Scheme A9 or A10, wherein the mutation status of the PIK3CA gene in the subject includes the absence of missense mutations at any one of codons 542, 545, 1047 and 1047.

[0431] Implementation Scheme A12. The method according to any one of Implementation Schemes A9-A11, wherein the mutational state of the PIK3CA gene in the subject includes the absence of a mutation at codon 542 that encodes an amino acid other than glutamic acid.

[0432] Implementation Scheme A13. The method according to any one of Implementation Schemes A9-A12, wherein the mutational state of the PIK3CA gene in the subject includes the absence of a mutation at codon 545 that encodes an amino acid other than glutamic acid.

[0433] Implementation Scheme A14. The method according to any one of Implementation Schemes A9-A13, wherein the mutational state of the PIK3CA gene in the subject includes the absence of a mutation at codon 1047 that encodes an amino acid other than histidine.

[0434] Implementation Scheme A15. The method according to any one of Implementation Schemes A9-A14, wherein the mutational state of the PIK3CA gene in the subject includes the absence of a mutation at codon 117 that encodes an amino acid other than lysine.

[0435] Implementation Scheme A16. The method according to any one of Implementation Schemes A9-A15, wherein the mutation status of the PIK3CA gene in the subject is the PIK3CA gene encoding the wild-type p110α protein.

[0436] Implementation Scheme A17. The method according to any one of Implementation Schemes A9-A16, wherein the mutation state of the PIK3CA gene in the subject is a state in which there is no copy number variation of the PIK3CA gene.

[0437] Implementation Scheme 19. The method according to any one of Implementation Schemes 9-17 and A1-A17, wherein the compound enhances the ability of the p53 mutant to bind DNA.

[0438] Implementation Scheme 20. The method according to any one of Implementation Schemes 9-18 and A1-A17, wherein the compound induces a conformational change in the p53 mutant.

[0439] Implementation Scheme 21. The method according to any one of Implementation Schemes 3-7 and A1-A17, wherein the compound selectively binds to the p53 mutant compared to wild-type p53.

[0440] Implementation Scheme 22. The method according to any one of Implementation Schemes 3, 5, 7 and A1-A17, wherein the mutation of the TP53 gene inactivates the p53 pathway in the subject.

[0441] Implementation Scheme 23. The method according to any one of Implementation Schemes 3, 5, 7, 22 and A1-A17, wherein the mutation of the TP53 gene is c.659A>G.

[0442] Implementation Scheme 24. The method according to any one of Implementation Schemes 3, 5, 7, 22, 23 and A1-A17, wherein the p53 mutant has the sequence according to SEQ ID NO: 3.

[0443] Implementation Scheme 25. The method according to any one of Implementation Schemes 3, 5, 7, 22-24 and A1-A17, wherein the mutation of the TP53 gene is at amino acid 220.

[0444] Implementation Scheme 26. The method according to any one of Implementation Schemes 3, 5, 7, 22-25 and A1-A17, wherein the TP53 gene contains a single nucleotide polymorphism.

[0445] Implementation Scheme 27. The method according to any one of Implementation Schemes 1-26 and A1-A17, wherein the p53 mutant is p53 Y220C.

[0446] Implementation Scheme 28. The method according to any one of Implementation Schemes 1-27 and A1-A17, wherein the p53 mutant has a reduced ability to bind to DNA in the subject compared to wild-type p53.

[0447] Implementation Scheme 29. The method according to any one of Implementation Schemes 1-28 and A1-A17, wherein the therapeutically effective amount is from about 50 mg to about 3000 mg.

[0448] Implementation Scheme 30. The method according to any one of Implementation Schemes 1-29 and A1-A17, wherein the application is once daily.

[0449] Implementation Scheme 31. The method according to any one of Implementation Schemes 2, 4, 5-7 and A1-A17, wherein the assay is DNA sequencing.

[0450] Implementation Scheme 32. The method according to any one of Implementation Schemes 2, 4, 5-7 and A1-A17, wherein the assay is next-generation DNA sequencing.

[0451] Implementation Scheme 33. The method according to any one of Implementation Schemes 2, 4, 5-7 and A1-A17, wherein the assay is RNA sequencing.

[0452] Implementation Scheme 34. The method according to any one of Implementation Schemes 2, 4, 5-7 and A1-A17, wherein the assay is a real-time polymerase chain reaction assay.

[0453] Implementation Scheme 35. The method according to any one of Implementation Schemes 2, 4, 5-7, 31-34 and A1-A17, wherein the biological sample is a liquid biopsy.

[0454] Implementation Scheme 36. The method according to any one of Implementation Schemes 2, 4, 5-7, 31-34 and A1-A17, wherein the biological sample is a blood sample.

[0455] Implementation Scheme 37. The method according to any one of Implementation Schemes 2, 4, 5-7, 31-34 and A1-A17, wherein the biological sample is circulating tumor DNA.

[0456] Implementation Scheme 38. The method according to any one of Implementation Schemes 2, 4, 5-7, 31-34 and A1-A17, wherein the biological sample is cell-free DNA.

[0457] Implementation Scheme 39. The method according to any one of Implementation Schemes 2, 4, 5-7, 31-34 and A1-A17, wherein the biological sample is a solid tumor sample.

[0458] Implementation Scheme 40. The method according to any one of Implementation Schemes 1-39 and A1-A17, wherein the condition is cancer.

[0459] Implementation Scheme 41. The method according to any one of Implementation Schemes 1-40 and A1-A17, wherein the condition is cancer, wherein the cancer expresses a p53 mutant.

[0460] Implementation Scheme 42. The method according to any one of Implementation Schemes 1-41 and A1-A17, wherein the condition is ovarian cancer.

[0461] Implementation Scheme 43. The method according to any one of Implementation Schemes 1-41 and A1-A17, wherein the condition is breast cancer.

[0462] Implementation Scheme 43A. The method according to Implementation Scheme 43, wherein the breast cancer is triple-negative breast cancer.

[0463] Implementation Scheme 44. The method according to any one of Implementation Schemes 1-41 and A1-A17, wherein the condition is lung cancer.

[0464] Implementation Scheme 44A. The method according to any one of Implementation Schemes 40-44 and 43A, wherein the cancer is HRD negative.

[0465] Implementation Scheme 45. The method according to any one of Implementation Schemes 1-44, 43A, 44A and A1-A17, wherein the administration is oral.

[0466] Implementation Scheme 46. The method according to any one of Implementation Schemes 1-44, 43A, 44A and A1-A17, wherein the administration is intravenous.

[0467] Implementation Scheme 47. The method according to any one of Implementation Schemes 1-44, 43A, 44A and A1-A17, wherein the application is subcutaneous.

[0468] Implementation Scheme 48. The method according to any one of Implementation Schemes 1-44, 43A, 44A and A1-A17, wherein the application is external.

[0469] Implementation Scheme 49. The method according to any one of Implementation Schemes 1-48, 43A, 44A and A1-A17, wherein the subject is a human.

[0470] Implementation Scheme 50. The method according to any one of Implementation Schemes 1-79, 43A, 44A and A1-A17, wherein said compound is: .

[0471] Implementation Scheme 51. The method according to any one of Implementation Schemes 1-79, 43A, 44A and A1-A17, wherein said compound is: .

[0472] Implementation Scheme 52. A method for treating cancer in a subject with a corresponding need, the method comprising: (a) Administering the subject a first course of treatment with a compound that binds to the p53 mutant; (b) Following (a), it is determined that the subject exhibited gastrointestinal adverse events; and (c) Based at least in part on determining that the subject exhibits the gastrointestinal adverse event, administer the compound, in a second course of treatment that is effective, to the subject along with food.

[0473] Implementation Scheme 53. The method according to Implementation Scheme 52, wherein the compound enhances the ability of the p53 mutant to bind DNA.

[0474] Implementation Scheme 84. The method according to Implementation Scheme 52 or 53, wherein the compound induces a conformational change in the p53 mutant.

[0475] Implementation Scheme 55. The method according to any one of Implementation Schemes 52-54, wherein the compound selectively binds to the p53 mutant compared to wild-type p53.

[0476] Implementation Scheme 56. The method according to any one of Implementation Schemes 52-55, wherein the compound is the compound according to any one of Implementation Schemes 50-81.

[0477] Implementation Scheme 57. The method according to any one of Implementation Schemes 52-56, wherein the cancer expresses the p53 mutant.

[0478] Implementation Scheme 58. The method according to any one of Implementation Schemes 52-57, wherein the p53 mutant is p53Y220C.

[0479] Implementation Scheme 59. The method according to any one of Implementation Schemes 52-58, wherein the cancer is ovarian cancer.

[0480] Implementation Scheme 60. The method according to any one of Implementation Schemes 52-58, wherein the cancer is breast cancer.

[0481] Implementation Scheme 61. The method according to any one of Implementation Schemes 52-58, wherein the cancer is lung cancer.

[0482] Implementation Scheme 62. The method according to any one of Implementation Schemes 52-61, wherein the first course of treatment is administered orally, and the second course of treatment is administered orally.

[0483] Implementation Scheme 63. The method according to any one of Implementation Schemes 52-62, wherein the gastrointestinal adverse events include nausea.

[0484] Implementation Scheme 64. The method according to any one of Implementation Schemes 52-62, wherein the gastrointestinal adverse events include diarrhea.

[0485] Implementation Scheme 65. The method according to any one of Implementation Schemes 52-62, wherein the gastrointestinal adverse event includes vomiting.

[0486] Implementation Scheme 66. The method according to any one of Implementation Schemes 52-65, wherein the gastrointestinal adverse events are self-reported by the subject.

[0487] Implementation Scheme 67. The method according to any one of Implementation Schemes 52-66, wherein the gastrointestinal adverse events are observed by a health practitioner.

[0488] Implementation Scheme 68. The method according to any one of Implementation Schemes 52-67, wherein administering the compound of the second course of treatment to the subject together with food includes having the subject eat before providing the compound to the subject.

[0489] Implementation Scheme 69. The method according to any one of Implementation Schemes 52-68, wherein administering the compound of the second course of treatment to the subject together with food includes having the subject eat approximately 5 minutes before providing the compound to the subject.

[0490] Implementation Scheme 70. The method according to any one of Implementation Schemes 52-67, wherein administering the compound of the second course of treatment to the subject together with food includes having the subject eat after providing the compound to the subject.

[0491] Implementation Scheme 71. The method according to any one of Implementation Schemes 52-67 and 70, wherein administering the compound of the second course of treatment to the subject together with food includes having the subject eat within approximately 60 minutes after providing the compound to the subject.

[0492] Implementation Scheme 72. The method according to any one of Implementation Schemes 52-67 and 70, wherein administering the compound of the second course of treatment to the subject together with food includes having the subject eat within approximately 30 minutes after providing the compound to the subject.

[0493] Implementation Scheme 73. The method according to any one of Implementation Schemes 52-67 and 70, wherein administering the compound of the second course of treatment to the subject together with food includes having the subject eat within approximately 15 minutes after providing the compound to the subject.

[0494] Implementation Scheme 74. The method according to any one of Implementation Schemes 52-67 and 70, wherein administering the compound of the second course of treatment to the subject together with food includes having the subject eat within approximately 5 minutes after providing the compound to the subject.

[0495] Implementation Scheme 75. The method according to any one of Implementation Schemes 52-74, wherein the food is a high-fat food.

[0496] Implementation Scheme 76. The method according to any one of Implementation Schemes 52-75, wherein the food contains at least 50% fat content of total calorie content.

[0497] Implementation Scheme 77. The method according to any one of Implementation Schemes 52-76, wherein the food contains at least 500 kcal of fat from fat.

[0498] Implementation Scheme 78. The method according to any one of Implementation Schemes 52-74, wherein the food is a medium-fat food.

[0499] Implementation Scheme 79. The method according to any one of Implementation Schemes 52-74 and 78, wherein the food contains about 25% to about 50% of the total calorie content as fat.

[0500] Implementation Scheme 80. The method according to any one of Implementation Schemes 52-74 and 78-79, wherein the food contains a fat content of about 125 kcal to about 500 kcal from fat.

[0501] Implementation Scheme 81. The method according to any one of Implementation Schemes 52-80, wherein the food is a high-calorie food.

[0502] Implementation Scheme 82. The method according to any one of Implementation Schemes 52-81, wherein the food contains at least 800 calories.

[0503] Implementation Scheme 83. The method according to any one of Implementation Schemes 52-82, wherein the first treatment course comprises a dose of the compound of about 50 mg to about 3000 mg, and the second treatment course comprises a dose of the compound of about 50 mg to about 3000 mg.

[0504] Implementation Scheme 84. The method according to any one of Implementation Schemes 52-82, wherein the first treatment course comprises a dose of about 1000 mg of the compound, and the second treatment course comprises a dose of about 1000 mg of the compound.

[0505] Implementation Scheme 85. The method according to any one of Implementation Schemes 52-82, wherein the first treatment course comprises a dose of about 2000 mg of the compound, and the second treatment course comprises a dose of about 2000 mg of the compound.

[0506] Implementation Scheme 86. The method according to any one of Implementation Schemes 52-82, wherein the first treatment course comprises a dose of about 3000 mg of the compound, and the second treatment course comprises a dose of about 3000 mg of the compound.

[0507] Implementation Scheme 87. A method for treating cancer, the method comprising: The following subjects were identified: (i) those who had been treated with a first course of the compound that bound the p53 mutant and were effective, and (ii) those who had experienced gastrointestinal adverse events after treatment with the first course of the compound. It is recommended that the compound be administered to the subject along with food, wherein the recommendation is based at least on the fact that the subject has already experienced the gastrointestinal adverse event; and According to the recommendations, the subject was given a second course of the compound, which was effective in treating the condition, along with food.

[0508] Implementation Scheme 88. The method according to Implementation Scheme 87, wherein the compound enhances the ability of the p53 mutant to bind DNA.

[0509] Implementation Scheme 89. The method according to any one of Implementation Schemes 87-88, wherein the compound induces a conformational change in the p53 mutant.

[0510] Implementation Scheme 90. The method according to any one of Implementation Schemes 87-89, wherein the compound selectively binds to the p53 mutant compared to wild-type p53.

[0511] Implementation Scheme 91. The method according to any one of Implementation Schemes 87-90, wherein the compound is the compound according to any one of Implementation Schemes 50-81.

[0512] Implementation Scheme 92. The method according to any one of Implementation Schemes 87-91, wherein the cancer expresses a p53 mutant.

[0513] Implementation Scheme 93. The method according to any one of Implementation Schemes 87-92, wherein the p53 mutant is p53Y220C.

[0514] Implementation Scheme 94. The method according to any one of Implementation Schemes 87-93, wherein the cancer is ovarian cancer.

[0515] Implementation Scheme 95. The method according to any one of Implementation Schemes 87-93, wherein the cancer is breast cancer.

[0516] Implementation Scheme 96. The method according to any one of Implementation Schemes 87-93, wherein the cancer is lung cancer.

[0517] Implementation Scheme 97. The method according to any one of Implementation Schemes 87-96, wherein the first course of treatment is administered orally, and the second course of treatment is administered orally.

[0518] Implementation Scheme 98. The method according to any one of Implementation Schemes 87-97, wherein the gastrointestinal adverse events include nausea.

[0519] Implementation Scheme 99. The method according to any one of Implementation Schemes 87-97, wherein the gastrointestinal adverse events include diarrhea.

[0520] Implementation Scheme 100. The method according to any one of Implementation Schemes 87-97, wherein the gastrointestinal adverse events include vomiting.

[0521] Implementation Scheme 101. The method according to any one of Implementation Schemes 87-100, wherein the gastrointestinal adverse events are self-reported by the subject.

[0522] Implementation Scheme 102. The method according to any one of Implementation Schemes 87-101, wherein the gastrointestinal adverse events are observed by a health practitioner.

[0523] Implementation Scheme 103. The method according to any one of Implementation Schemes 87-102, wherein administering the compound of the second course of treatment to the subject together with food includes having the subject eat before providing the compound to the subject.

[0524] Implementation Scheme 104. The method according to any one of Implementation Schemes 87-103, wherein administering the compound of the second course of treatment to the subject together with food includes having the subject eat approximately 5 minutes before providing the compound to the subject.

[0525] Implementation Scheme 105. The method according to any one of Implementation Schemes 87-102, wherein administering the compound of the second course of treatment to the subject together with food includes having the subject eat after providing the compound to the subject.

[0526] Implementation Scheme 106. The method according to any one of Implementation Schemes 87-102 and 105, wherein administering the compound of the second course of treatment to the subject together with food includes having the subject eat within approximately 60 minutes after providing the compound to the subject.

[0527] Implementation Scheme 107. The method according to any one of Implementation Schemes 87-102 and 105, wherein administering the compound of the second course of treatment to the subject together with food includes having the subject eat within approximately 30 minutes after providing the compound to the subject.

[0528] Implementation Scheme 108. The method according to any one of Implementation Schemes 87-102 and 105, wherein administering the compound of the second course of treatment to the subject together with food includes having the subject eat within approximately 15 minutes after providing the compound to the subject.

[0529] Implementation Scheme 109. The method according to any one of Implementation Schemes 87-102 and 105, wherein administering the compound of the second course of treatment to the subject together with food includes having the subject eat within approximately 5 minutes after providing the compound to the subject.

[0530] Implementation Scheme 110. The method according to any one of Implementation Schemes 87-109, wherein the food is a high-fat food.

[0531] Implementation Scheme 111. The method according to any one of Implementation Schemes 87-110, wherein the food contains at least 50% fat content of total calorie content.

[0532] Implementation Scheme 112. The method according to any one of Implementation Schemes 87-111, wherein the food contains at least 500 kcal of fat from fat.

[0533] Implementation Scheme 113. The method according to any one of Implementation Schemes 87-109, wherein the food is a medium-fat food.

[0534] Implementation Scheme 114. The method according to any one of Implementation Schemes 87-109 and 113, wherein the food contains about 25% to about 50% of the total calorie content as fat.

[0535] Implementation Scheme 115. The method according to any one of Implementation Schemes 87-109 and 113-114, wherein the food contains a fat content of about 125 kcal to about 500 kcal from fat.

[0536] Implementation Scheme 116. The method according to any one of Implementation Schemes 87-115, wherein the food is a high-calorie food.

[0537] Implementation Scheme 117. The method according to any one of Implementation Schemes 87-116, wherein the food contains at least 800 calories.

[0538] Implementation Scheme 118. The method according to any one of Implementation Schemes 87-117, wherein the first treatment course comprises a dose of the compound of about 50 mg to about 3000 mg, and the second treatment course comprises a dose of the compound of about 50 mg to about 3000 mg.

[0539] Implementation Scheme 119. The method according to any one of Implementation Schemes 87-117, wherein the first treatment course comprises a dose of about 1000 mg of the compound, and the second treatment course comprises a dose of about 1000 mg of the compound.

[0540] Implementation Scheme 120. The method according to any one of Implementation Schemes 87-117, wherein the first treatment course comprises a dose of about 2000 mg of the compound, and the second treatment course comprises a dose of about 2000 mg of the compound.

[0541] Implementation Scheme 121. The method according to any one of Implementation Schemes 87-117, wherein the first treatment course comprises a dose of about 3000 mg of the compound, and the second treatment course comprises a dose of about 3000 mg of the compound.

[0542] Implementation Scheme 122. The method according to any one of Implementation Schemes 52-121, wherein said compound is: .

[0543] Implementation Scheme 123. The method according to any one of Implementation Schemes 52-121, wherein said compound is: .

Claims

1. A method of treating a condition in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a small molecule compound that binds a p53 mutant protein, wherein the mutational status of the KRAS gene in the subject has been previously determined.

2. The method of claim 1, wherein the mutational status of the KRAS gene in the subject has been previously determined based on an assay performed on a biological sample of the subject.

3. The method of claim 1 or claim 2, wherein the subject has been previously determined to have a mutation in the TP53 gene encoding a p53 mutant in the subject.

4. A method of treating a condition in a subject in need thereof, the method comprising: a) determining the mutational status of the KRAS gene in the subject based on the results of an assay performed on a biological sample of the subject; and b) administering to the subject a therapeutically effective amount of a compound that binds a p53 mutant protein based at least in part on the mutational status of the KRAS gene in the subject.

5. The method of claim 4, wherein the method further comprises determining that the subject carries a mutation in the TP53 gene encoding a p53 mutant protein based on the results of the assay, wherein administering to the subject the therapeutically effective amount of a compound that binds a p53 mutant protein is based at least in part on determining that the subject carries a mutation in the TP53 gene encoding the p53 mutant protein.

6. The method of any one of claims 2, 4, and 5, further comprising performing the assay on the biological sample of the subject.

7. The method of claim 5, further comprising performing the assay on the biological sample of the subject, wherein performing the assay on the biological sample of the subject further comprises detecting a mutation in the TP53 gene encoding the p53 mutant in the subject.

8. The method of any one of claims 1-7, wherein the mutational status of the KRAS gene in the subject comprises the absence of a missense mutation at any one of codons 12, 13, 59, 61, 117, and 146.

9. The method of any one of claims 1-8, wherein the mutational status of the KRAS gene in the subject comprises the absence of a mutation at codon 12 that would encode an amino acid other than glycine.

10. The method of any one of claims 1-9, wherein the mutational status of the KRAS gene in the subject comprises the absence of a mutation at codon 13 that would encode an amino acid other than glycine.

11. The method of any one of claims 1-10, wherein the mutational status of the KRAS gene in the subject comprises the absence of a mutation at codon 59 that would encode an amino acid other than alanine.

12. The method of any one of claims 1-11, wherein the mutational status of the KRAS gene in the subject comprises the absence of a mutation at codon 61 that would encode an amino acid other than glutamine.

13. The method of any one of claims 1-12, wherein the mutational status of the KRAS gene in the subject comprises the absence of a mutation at codon 117 that would encode an amino acid other than lysine.

14. The method of any one of claims 1-13, wherein the mutational status of the KRAS gene in the subject comprises the absence of a mutation at codon 146 that would encode an amino acid other than alanine.

15. The method of any one of claims 1-14, wherein the mutational status of the KRAS gene in the subject is a KRAS gene that encodes a wild-type K-ras protein.

16. The method of claim 15, wherein the wild-type K-Ras protein has a sequence according to SEQ ID NO: 1 or SEQ ID NO:

2.

17. The method of any one of claims 1-16, wherein the mutational status of the KRAS gene in the subject is a copy number variation status of the KRAS gene.

18. The method of claim 17, wherein the mutational status of the KRAS gene in the subject is the absence of a KRAS copy number variation.

19. The method of any one of claims 1-17, wherein the compound enhances the ability of a p53 mutant to bind DNA.

20. The method of any one of claims 1-18, wherein the compound induces a conformational change in a p53 mutant.

21. The method of any one of claims 3-7, wherein the compound selectively binds to a p53 mutant as compared to wild-type p53.

22. The method of any one of claims 3, 5, and 7, wherein the mutation of the TP53 gene inactivates the p53 pathway in the subject.

23. The method of any one of claims 3, 5, 7, and 22, wherein the mutation of the TP53 gene is c.659A>G.

24. The method of any one of claims 3, 5, 7, 22, and 23, wherein the p53 mutant has a sequence according to SEQ ID NO:

3.

25. The method of any one of claims 3, 5, 7, and 22-24, wherein the mutation of the TP53 gene is at amino acid 220.

26. The method of any one of claims 3, 5, 7, and 22-25, wherein the TP53 gene contains a single nucleotide polymorphism.

27. The method of any one of claims 1-26, wherein the p53 mutant is p53 Y220C.

28. The method of any one of claims 1-27, wherein the p53 mutant has a reduced ability to bind to DNA in the subject as compared to wild-type p53.

29. The method of any one of claims 1-28, wherein the therapeutically effective amount is from about 50 mg to about 3000 mg.

30. The method of any one of claims 1-29, wherein the administering is once a day.

31. The method of any one of claims 2, 4, and 5-7, wherein the assay is DNA sequencing.

32. The method of any one of claims 2, 4, and 5-7, wherein the assay is next generation DNA sequencing.

33. The method of any one of claims 2, 4, and 5-7, wherein the assay is RNA sequencing.

34. The method of any one of claims 2, 4, and 5-7, wherein the assay is a real-time polymerase chain reaction assay.

35. The method of any one of claims 2, 4, 5-7, and 31-34, wherein the biological sample is a liquid biopsy.

36. The method of any one of claims 2, 4, 5-7, and 31-34, wherein the biological sample is a blood sample.

37. The method of any one of claims 2, 4, 5-7, and 31-34, wherein the biological sample is circulating tumor DNA.

38. The method of any one of claims 2, 4, 5-7, and 31-34, wherein the biological sample is cell-free DNA.

39. The method of any one of claims 2, 4, 5-7, and 31-34, wherein the biological sample is a solid tumor sample.

40. The method of any one of claims 1-39, wherein the condition is cancer.

41. The method of any one of claims 1-40, wherein the condition is cancer, wherein the cancer expresses a p53 mutant.

42. The method of any one of claims 1-41, wherein the condition is ovarian cancer.

43. The method of any one of claims 1-41, wherein the condition is breast cancer.

44. The method of any one of claims 1-41, wherein the condition is lung cancer.

45. The method of any one of claims 1-44, wherein the administration is oral.

46. The method of any one of claims 1-44, wherein the administration is intravenous.

47. The method of any one of claims 1-44, wherein the administration is subcutaneous.

48. The method of any one of claims 1-44, wherein the administration is topical.

49. The method of any one of claims 1-48, wherein the subject is a human.

50. The method of any one of claims 1-49, wherein the compound is: 。 51. The method of any one of claims 1-49, wherein the compound is: 。

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