Use of a PI3Ka inhibitor in the preparation of a medicament for treating a disease associated with a mutation in the PIK3CA or / and TEK gene

Abstract

The present application provides use of a PI3Kα inhibitor in the manufacture of a medicament for treating a disease associated with a PIK3CA or / and TEK gene mutation. In particular, the present application provides use of a PI3Kα inhibitor CYH33 or a pharmaceutical composition comprising CYH33 in the manufacture of a medicament for preventing, treating or alleviating a PIK3CA-related overgrowth syndrome (PROS) and / or a PIK3CA-related vascular malformation (PRVM), and / or a TEK gene mutation-related vascular malformation.

Description

Technical Field

[0001] This invention relates to novel uses of PI3Kα inhibitors, and more particularly to the use of CYH33 in the preparation of medicaments for the prevention or treatment of disorders or diseases associated with PIK3CA and / or TEK gene mutations. Background Technology

[0002] PIK3CA, encoding the catalytic subunit of phosphatidylinositol-3-kinase (PI3K), is an oncogene. Activating mutations in PIK3CA, in addition to being associated with the development of various malignant tumors, also trigger a series of other PIK3CA-related diseases (especially PIK3CA-related malformations), including PIK3CA-associated overgrowth syndrome (PROS) and PIK3CA-associated vascular malformations (PRVM). Among PIK3CA-related diseases, hotspot mutations (p.E542K, p.E545K, p.H1047R, and p.H1047L) found in tumors also have a high incidence in non-tumor diseases; furthermore, non-hotspot mutations, including p.C420R, p.E726K, and p.G914R, have also been shown to occur at relatively high frequencies. Tyrosine kinase 2 (TIE2), an endothelial-specific type I transmembrane protein receptor tyrosine kinase encoded by the TEK gene, mediates angiopoietin signaling, thereby promoting endothelial cell survival, vascular remodeling, and integrity. Genetic or somatic variations of TEK, such as G833D, Q837H, Y897S / H / C, L914F, R915C, R918C / H, and K1100N, have been identified as being associated with the pathogenesis of hereditary or sporadic venous malformations. TEK mutations enable ligand-independent autophosphorylation, leading to overactivation of downstream pathways such as PI3K / AKT.

[0003] PROS is a collective term for a series of PIK3CA-related overgrowth disorders. Its pathogenesis is caused by activating mutations in the oncogene PIK3CA, leading to abnormal activation of the PI3K / AKT / mTOR signaling pathway. This results in segmental, asymmetric, or invasive growth of tissues in various parts of the body, with or without vascular lesions, leading to rare diseases of overgrowth in multiple sites, such as fibrofatty hyperplasia or overgrowth (FAO), fibrofatty infiltrative lipomatosis / facial infiltrative lipomatosis, macrodactyly, dysplastic megalencephaly (DMEG), unilateral hyperplasia with multiple lipomatosis (HHML), megalencephaly-capillary malformation (M-CM), Klippel-Trenaunay syndrome, Cloves syndrome, and a series of other related rare syndromes.

[0004] Vascular abnormalities can be broadly classified into hemangiomas and vascular malformations. This classification system was first proposed in 1982, accepted by the International Society for the Study of Vascular Malformations in 1996, and revised again in 2018. Vascular malformations mainly include simple vascular malformations, mixed vascular malformations, and other types. Among them, simple vascular malformations can be further divided into capillary malformations (CM), lymphatic malformations (LM), venous malformations (VM), arteriovenous malformations (AVM), and arteriovenous fistulas (AVF), etc.; LM includes ordinary (cystic) lymphatic malformations, generalized lymphatic malformations (GLA), and lymphatic malformations in Gorham-Stout syndrome (GSD), etc.; VM includes ordinary venous malformations, familial mucocutaneous venous malformations (VMCM), and blue rubber nevus (Bean) syndrome (BRBN syndrome), etc. The causes of vascular malformations are multifaceted, including mutations in the TEK gene encoding the endothelial receptor tyrosine kinase TIE2, mutations in the oncogene PIK3CA, a combination of TEK and PIK3CA mutations, or other mutations. Vascular malformations caused by mutations in the oncogene PIK3CA are called PIK3CA-associated vascular malformations (PRVMs). Patients present with varying clinical manifestations, depending on the affected site, and are predominantly pediatric. It has been reported that over 80% of lymphangiomas (LMs) carry PIK3CA mutations, with the majority of LMs being common (cystic) lymphangiomas; over 55% of gamma-lymphatic lesions (GLAs) carry PIK3CA mutations. Approximately 20% of VMs are associated with PIK3CA mutations, and approximately 50%-70% of VMs are caused by TEK gene mutations. Based on the type of TEK mutation, venous malformations (VMs) can be further classified into single venous malformations (TEK L914F somatic mutation), multiple venous malformations (TEK R915C chimeric mutation combined with TEK Y897C somatic mutation), VMCM (TEK R849W germline mutation combined with TEK Y1108 somatic mutation), and blue rubber nevus syndrome (BRBNS) (TEK T1105NT1106P somatic double mutation). Among these, the TEK L914F somatic mutation is the most common mutation (accounting for approximately 60%).

[0005] Due to the diverse and complex nature of PROS / PRVM, there is currently no standard treatment. Treatment strategies are typically individualized, including surgical resection, laser resection, and sclerotherapy. Based on the severity and urgency of disease progression, treatment strategies can be broadly categorized into three types: i) Asymptomatic lesions on the body surface without enlargement can be closely monitored; ii) Symptomatic lesions on the body surface or those with enlargement, as well as all visceral lesions, require aggressive treatment. Since lesions that can be completely resected without functional impairment are extremely rare, systemic medication should be the first choice; iii) For patients with severe complications, such as Kaposi's hemangioendothelioma (KHE) patients with coagulation disorders, anticoagulants and blood products should be administered in addition to aggressive tumor treatment to improve coagulation function. Surgical resection or laser resection is usually the preferred treatment for PROS and PRVM, but the boundaries between PROS and PRVM lesions are often unclear, frequently involving important blood vessels, nerves, and even organs, greatly increasing surgical risks and the possibility of functional impairment. Extensive excision often results in severe scarring post-surgery, and can sometimes lead to shock, death, or other serious consequences due to severe bleeding. Furthermore, the recurrence rate is high. Sclerotherapy is primarily used for patients with vascular malformations and is ineffective for limb overgrowth and deformities.

[0006] Sirolimus, an inhibitor of mTOR, a downstream protein of the PI3K / AKT pathway, has been reported to have some efficacy in patients with proliferative malformations (PROS) and can improve vascular malformations. However, sirolimus does not directly target gene mutations such as PIK3CA or TEK, so patients are prone to drug resistance or relapse after a period of treatment, and its long-term use is limited due to side effects. Alpelisib (BYL719), a PI3Kα inhibitor, is the world's first and only approved treatment for PROS, for use in adults and children aged 2 years and older with severe clinical manifestations of PROS requiring systemic therapy. Currently, no targeted therapies are approved for the treatment of PRVM and TEK gene mutation-related vascular malformations. Overall, patients with PIK3CA and / or TEK gene mutation-related growth malformations and / or vascular malformations urgently need systemically treatable, effective, and safe therapeutic agents. Summary of the Invention

[0007] This invention relates to the use of the PI3Kα inhibitor CYH33 in the prevention, treatment, or alleviation of PIK3CA-related malformations, particularly PIK3CA-related overgrowth syndrome (PROS) and / or PIK3CA-related vascular malformations (PRVM), and / or TEK gene mutation-related vascular malformations, especially venous or lymphatic malformations. The inventors have found that the PI3Kα inhibitor CYH33 has good therapeutic effects on subjects carrying PROS and / or PRVM, subjects carrying TEK gene mutation-related vascular malformations, and subjects carrying both PIK3CA gene mutations and TEK gene mutation-related vascular malformations.

[0008] On the one hand, the present invention provides the use of PI3Kα inhibitor CYH33 or pharmaceutical compositions containing CYH33 in the preparation of medicaments for the prevention, treatment or relief of PIK3CA-related malformations, especially PIK3CA-related overgrowth syndrome (PROS) and / or PIK3CA-related vascular malformations (PRVM), and / or TEK gene mutation-related vascular malformations.

[0009] In one embodiment, the present invention provides the use of the PI3Kα inhibitor CYH33 or a pharmaceutical composition containing CYH33 in the preparation of a medicament for the prevention, treatment or relief of vascular malformations associated with PROS, PRVM, or TEK gene mutations.

[0010] In one embodiment, the present invention provides the use of the PI3Kα inhibitor CYH33 or a pharmaceutical composition containing CYH33 in the preparation of a medicament for the prevention, treatment or relief of PROS and / or PRVM.

[0011] In one embodiment, the present invention provides the use of the PI3Kα inhibitor CYH33 or a pharmaceutical composition containing CYH33 in the preparation of a medicament for the prevention, treatment or relief of PROS and PRVM.

[0012] In one embodiment, the present invention provides the use of the PI3Kα inhibitor CYH33 or a pharmaceutical composition containing CYH33 in the preparation of a medicament for the prevention, treatment or mitigation of TEK gene mutation-related vascular malformations.

[0013] In one embodiment, the present invention provides the use of the PI3Kα inhibitor CYH33 or a pharmaceutical composition containing CYH33 in the preparation of a medicament for the prevention, treatment or relief of PIK3CA-related malformations and TEK gene mutation-related vascular malformations.

[0014] On the other hand, the present invention provides a method for preventing, treating or alleviating PIK3CA-related malformations, particularly PIK3CA-related overgrowth syndrome (PROS) and / or PIK3CA-related vascular malformations (PRVM), and / or TEK gene mutation-related vascular malformations, the method comprising administering an effective amount of CYH33 or a pharmaceutical composition containing CYH33 to a subject in need of such treatment.

[0015] In one embodiment, the present invention provides a method for preventing, treating, or alleviating PROS and / or PRVM, the method comprising administering a therapeutically effective amount of CYH33 or a pharmaceutical composition containing CYH33 to a subject in need of such treatment.

[0016] In one embodiment, the present invention provides a method for preventing, treating, or alleviating PROS and PRVM, the method comprising administering a therapeutically effective amount of CYH33 or a pharmaceutical composition containing CYH33 to a subject in need of such treatment.

[0017] In one embodiment, the present invention provides a method for preventing, treating, or mitigating TEK gene mutation-related vascular malformations, the method comprising administering a therapeutically effective amount of CYH33 or a pharmaceutical composition containing CYH33 to a subject in need of such treatment.

[0018] In one embodiment, the present invention provides a method for preventing, treating, or mitigating PIK3CA-related malformations and TEK gene mutation-related vascular malformations, the method comprising administering a therapeutically effective amount of CYH33 or a pharmaceutical composition containing CYH33 to a subject in need of such treatment.

[0019] In some embodiments of the present invention, the subject carries a PIK3CA gene mutation and / or a TEK gene mutation.

[0020] In some embodiments of the invention, the subject carries PIK3CA-associated overgrowth syndrome (PROS) or PIK3CA-associated vascular malformation (PRVM).

[0021] In some embodiments of the invention, the subject carries PIK3CA-associated overgrowth syndrome (PROS) and PIK3CA-associated vascular malformation (PRVM).

[0022] In some embodiments of the present invention, the subject carries a TEK gene mutation-related vascular malformation.

[0023] In some embodiments of the present invention, the subject carries PROS disease and / or PRVM disease, and TEK gene mutation-related vascular malformation disease.

[0024] In some embodiments of the present invention, the subject carries TEK gene mutation-related vascular malformations and PIK3CA-related malformations (especially vascular malformations).

[0025] In some embodiments, the pharmaceutical composition includes a therapeutically effective amount of CYH33, and pharmaceutically acceptable excipients or carriers, etc.

[0026] In some embodiments, the pharmaceutical composition is formulated for intravenous, intramuscular, oral, rectal, inhalation, nasal, topical, ocular, or ocular administration.

[0027] In some embodiments, the pharmaceutical composition is a tablet, pill, capsule, liquid, inhaler, nasal spray solution, suppository, solution, emulsion, ointment, eye drop, or ear drop.

[0028] In some embodiments of the present invention, the PROS include, but are not limited to, a series of rare syndromes such as macrodactyly / toe, Cloves syndrome, Clappo syndrome, fibrofatty hyperplasia or overgrowth (FAO), Klippel-Trenaunay syndrome, unilateral hyperplastic multiple lipomatosis (HHML), macrocephaly-capillary malformation (MCAP), diffuse capillary malformation with overgrowth (DCMO), fibrofatty vascular lesion (FAVA), facial invasive lipoma (FIL), dysplastic macrocephaly (DMEG), hemilateral macrocephaly (HMEG), muscular hyperplasia (Muscular HH), seborrheic keratosis (SK), and benign lichenoid keratosis (BLK). Preferably, the PROS are selected from a series of related rare syndromes such as Cloves syndrome, Klippel-Trenaunay syndrome, FAO, HHML, and MCAP.

[0029] In some embodiments of the present invention, the PRVM includes, but is not limited to: simple vascular malformations (such as capillary malformations (CM), venous malformations (VM), lymphatic malformations (LM)) and mixed vascular malformations.

[0030] In some embodiments of the present invention, lymphatic malformations (LM) mainly include common (cystic) lymphatic malformations (such as giant cystic lymphatic malformations, microcystic lymphatic malformations, and mixed cystic lymphatic malformations), generalized lymphatic malformations (GLA) (such as Kaposi's lymphangiomatosis (KLA)), lymphatic malformations in Gorham-Stout syndrome (GSD), tubular lymphatic malformations, and a series of rare syndromes such as lymphedema.

[0031] In some embodiments of the present invention, capillary malformations (CM) mainly include simple vascular nevi / salmon spots, skin and / or mucous membrane CM, reticular capillary malformations, capillary malformations in capillary malformations-arteriovenous malformations (CM-AVM), congenital telangiectatic marbled skin (CMTC), telangiectasia, etc.

[0032] In some embodiments of the present invention, venous malformations (VMs) mainly include common venous malformations, etc.

[0033] In a preferred embodiment of the present invention, PRVM mainly includes capillary malformations (such as reticular capillary malformations, capillary malformations in capillary malformation-arteriovenous malformation (CM-AVM), etc.), lymphatic malformations (such as ordinary (cystic) lymphatic malformations, generalized lymphatic malformations (GLA), etc.), venous malformations such as ordinary venous malformations (VM), and mixed vascular malformations (such as capillary-venous malformations (CVM), capillary-lymphatic malformations (CLM), lymphatic-venous malformations (LVM), capillary-lymphatic-venous malformations (CLVM), etc.).

[0034] In some embodiments of the present invention, the TEK gene mutation-related vascular malformation is a venous malformation or a lymphatic malformation, etc.; preferably, it is a venous malformation.

[0035] In some embodiments of the present invention, TEK gene mutation-related vascular malformations (especially venous malformations) include, but are not limited to: single venous malformation, multiple venous malformations, familial mucocutaneous venous malformation (VMCM), and blue rubber nipple nevus (Bean) syndrome (BRBN syndrome).

[0036] In some embodiments of the present invention, the PIK3CA gene mutation and TEK gene mutation-related vascular malformations include, but are not limited to, venous malformations.

[0037] Terminology Explanation

[0038] In this invention, unless otherwise expressly stated, the terminology used herein has the meanings defined below. Terms not explicitly defined in this invention have their general meanings as commonly understood by those skilled in the art.

[0039] As used herein, the term "CYH33" refers to a highly selective PI3Kα inhibitor that exhibits significant inhibitory activity against PI3Kα and its mutants, particularly wild-type and mutant PI3Kα kinases. Its chemical name is methyl 5-{6-[(4-methanesulfonylpiperazin-1-yl)methyl]-4-morpholinylpyrrole[2,1-f][1,2,4]triazin-2-yl}-4-trifluoromethylpyridin-2-yl}carbamate.

[0040] "Fibrolipogenic hyperplasia or overgrowth (FAO)" refers to a syndrome characterized by segmental, progressive overgrowth of subcutaneous, muscular, and visceral fibrolipid tissues with skeletal overgrowth.

[0041] "Megabrain-capillary malformation (M-CM) syndrome" refers to the syndrome with the following main findings: (1) megalencephaly (MEG) or hemimegalencephaly (HMEG) associated with neurological findings of hypotonia, epilepsy and mild to severe intellectual disability; and (2) cutaneous capillary malformations with focal or generalized overgrowth of the body.

[0042] "CLOVES syndrome" refers to congenital, lipid, overgrowth, vascular malformations, epidermal nevi, and spinal / skeletal abnormalities and / or scoliosis. The syndrome is characterized by a complex combination of congenital overgrowth of lipid tissue (typically manifesting as a lipid mass in the trunk) and vascular and lymphatic malformations.

[0043] Klippel-Trenaunay syndrome is a rare congenital medical condition in which blood vessels and / or lymphatic vessels cannot form properly.

[0044] As used herein, the term “treatment” refers to preventative or therapeutic treatment, as well as curative or disease-modifying treatment, including treatment of patients at risk of contracting the disease or suspected of having contracted the disease, and subjects who are ill or diagnosed with a disease or medical condition, including suppression of clinical relapse. Treatment may be administered to subjects with a medical condition or who may eventually contract the disease in order to prevent, cure, delay the onset of one or more symptoms of the disease or relapse, reduce the severity of one or more symptoms of the disease or relapse, or alleviate one or more symptoms of the disease or relapse, or to extend the subject’s survival beyond what would be expected without such treatment. “Treatment regimen” refers to a mode of treatment for the disease, such as a dosage pattern used during treatment. Treatment regimens may include induction regimens and maintenance regimens. The phrase “induction regimen” or “induction period” refers to a treatment regimen (or part of a treatment regimen) used for the initial treatment of the disease. The overall goal of an induction regimen is to provide the subject with a high level of the drug at the initial stage of the treatment regimen. An induction regimen may employ a (partial or complete) “loading regimen,” which may include administering a larger dose of the drug than a physician would use during a maintenance regimen, such as administering the drug more frequently as a physician would during a maintenance regimen, or both. The phrase "maintenance regimen" or "maintenance period" refers to a treatment regimen (or part of a treatment regimen) used to maintain a subject's remission for a long period (months or years) during the treatment of a disease. Maintenance regimens can be continuous treatment (e.g., administration of medication at regular intervals, such as weekly, monthly, or yearly) or intermittent treatment (e.g., interrupted treatment, intermittent treatment, treatment during relapse, or treatment to achieve specific predetermined criteria).

[0045] The term "therapeutic effective amount" refers to the amount of the compound of the present invention that can elicit a biological or medical response in an individual or subject, or improve symptoms, slow or delay the progression of a disease, or prevent the disease.

[0046] As used herein, the term "individual or subject" refers to an animal. Preferably, the animal is a mammal. "Individual or subject" also refers to, for example, primates (e.g., humans), cattle, sheep, goats, horses, dogs, cats, rabbits, rats, mice, fish, birds, etc. In a preferred embodiment, the individual or subject is a human.

[0047] As used herein, the term “inhibition” refers to the reduction or suppression of a specific patient, symptom, condition, or disease, or a significant reduction in biological activity or baseline activity of a process.

[0048] As used herein, in one embodiment, the term "treatment" for any disease or condition refers to improving the disease or condition (i.e., stopping or slowing the development of the disease or at least one of its clinical symptoms). In another embodiment, "treatment" refers to improving at least one bodily parameter, which may not be perceptible to the patient. In yet another embodiment, "treatment" refers to regulating the disease or condition physically (e.g., stabilizing perceptible symptoms) or physiologically (e.g., stabilizing bodily parameters) or both.

[0049] Beneficial effects

[0050] The PI3Kα inhibitor CYH33 showed good therapeutic effects in PROS and / or PRVM subjects carrying PIK3CA gene mutations, as well as subjects with venous malformations caused by TEK gene mutations. CYH33's ability to inhibit the proliferation of PIK3CA-mutated and TEK-mutated angiocytes and its ability to inhibit the growth of mutant transplanted tissues were significantly stronger than BYL719. Furthermore, in a Phase I first-in-human clinical trial, the types of adverse events (AEs) with CYH33 were similar to those with BYL719, but the incidence of some AEs (such as abnormal liver enzymes and diarrhea) was significantly lower than that with BYL719. In addition, CYH33 exhibited good safety and pharmacokinetic characteristics at QD levels of 5, 10, and 15 mg, with outstanding efficacy; 87.5% of patients experienced a reduction in target lesion volume of at least 20%, and some even saw a reduction of 30-65%.

[0051] The present invention is further illustrated by the following figures and embodiments. However, these embodiments and figures should not be construed as limiting the scope of the invention in any way. Attached Figure Description

[0052] Figure 1 This is a graph showing the effect of compound CYH33 in Example 1 on the volume of transplanted tissue in a nude mouse model with subcutaneous allogeneic transplantation of NIH / 3T3-PIK3CAH1047R cells;

[0053] Figure 2 This is a graph showing the effect of compound CYH33 in Example 1 on the change in body weight in mice in the NIH / 3T3-PIK3CAH1047R cell subcutaneous allogeneic transplantation model.

[0054] Figure 3 This is a graph showing the effect of compound CYH33 in Example 2 on inhibiting the in vitro proliferation of HMEC-1-PIK3CA-H1047R cells;

[0055] Figure 4 This is a graph showing the effect of compounds CYH33 and BYL719 in Example 3 on the volume of transplanted tissue in a nude mouse model with subcutaneous allogeneic transplantation of NIH / 3T3-FL-TIE2-L914F cells;

[0056] Figure 5 This is a graph showing the effects of compounds CYH33 and BYL719 in Example 3 on the changes in body weight in mice using the NIH / 3T3-FL-TIE2-L914F cell subcutaneous allogeneic transplantation model.

[0057] Figure 6 This is a graph showing the inhibition of HMEC-1-TIE2-L914F cell proliferation in vitro by compounds CYH33 and BYL719 in Example 4;

[0058] Figure 7 The graph shows the effect of compound CYH33 on the phosphorylation level of AKT after 2 hours of treatment of cells in Example 5 (a) and the gray value analysis of the protein band (b).

[0059] Figure 8 This is a graph showing the change in target lesion volume in adult PROS / PRVM patients after receiving different doses of CYH33 treatment in Example 6. Detailed Implementation

[0060] The following embodiments illustrate the present invention as described above; however, they do not limit the scope of this disclosure in any way. The beneficial effects of combinations of the present disclosure can also be determined using other test models known to those skilled in the art.

[0061] Experimental methods for in vitro cell viability studies

[0062] Cells in the logarithmic growth phase were used for plating. The cell concentration was adjusted, and 90 μL of cell suspension was added to each well of the culture plate. Cell-free culture medium was added to the blank control wells. The culture plates were incubated overnight at 37°C, 5% CO2, and 100% relative humidity. 10 μL of different concentrations of CYH33 working solution were added to the cell culture plates, with two replicates per group. 10 μL of DMSO-cell culture medium mixture was added to the solvent control (containing only cells and cell culture medium) and the blank control (containing cell culture medium but no cells). The final concentration of DMSO was 0.1%. The 96-well cell plate was returned to the incubator and cultured for 7 days. Then, add 50 μL (equivalent to half the volume of cell culture medium in each well) of CellTiter-Glo working solution from the Promega CellTiter-Glo luminescent cell viability assay kit (Promega-G7573) to each well, wrap the cell plate with aluminum foil to protect it from light, shake the culture plate on a track shaker for 2 minutes to induce cell lysis, and place the culture plate at room temperature for 10 minutes to stabilize the luminescence signal. Detect the luminescence signal (expressed as RLU (relative light units)) using a microplate reader.

[0063] The inhibition rate (IR0) of the test compound was calculated using the following formula in in vitro cell viability assays: IR0(%) = (1 - (RLU compound - RLU blank control) / (RLU solvent control - RLU blank control)) × 100%. The inhibition rates of different compound concentrations were calculated in Excel, and then the inhibition curve was fitted using log(inhibitor) vs. response – variable slope software in GraphPad Prism 8.0 to obtain relevant parameters, including minimum inhibition rate, maximum inhibition rate, and relative IC50. 50 .

[0064] The minimum inhibition rate (MIC) refers to the Y value corresponding to the bottom plateau of the curve, while the maximum inhibition rate (MPR) is the Y value corresponding to the top plateau of the curve, relative to the IC. 50 It is the concentration required to bring the curve down to the point halfway between the top and bottom plateaus of the curve, absolute IC. 50 This refers to the drug concentration at which cell viability is inhibited by half.

[0065] In vivo drug efficacy experiments: Measurement of transplanted tissue and animal body weight.

[0066] The effectiveness of treatment was assessed based on the growth of the transplanted tissue. Once the transplanted tissue was palpable, its volume and size were evaluated twice weekly. The volume and size of the transplanted tissue were determined using a digital caliper, and the animal's weight was measured twice weekly throughout the trial. The weight of the transplanted tissue was measured at the end of treatment.

[0067] Transplanted tissue volume V in mm 3 This is expressed as follows: V = 0.5a × b 2 , where a and b are the long and short diameters of the transplanted tissue, respectively.

[0068] T / C and TGI are indicators reflecting the response of transplanted tissue volume to treatment. T / C (%) reflects the relative proliferation rate of transplanted tissue (volume), i.e., the percentage of treatment / control (T / C) value of the transplanted tissue, calculated using the following formula:

[0069] T / C (%) = (T RTV / C RTV )×100,(T RTV C RTV The values ​​represent the average relative transplanted tissue volume (RTV) on the day of treatment for the treatment group and the solvent control group, respectively. The relative transplanted tissue volume (RTV) was calculated based on the transplanted tissue measurements using the formula: RTV = Vt / V0, where V0 is the average transplanted tissue volume measured at the start of treatment (i.e., day 0), Vt is the average transplanted tissue volume measured after a certain treatment time (t), and T...RTV With C RTV Take data from the same day.

[0070] TGI (%) reflects the growth inhibition rate of transplanted tissue. The formula for calculating TGI (%) is: TGI (%) = [1 - (T i -T0) / (V i -V0)]×100. Where, T i T0 and T0 represent the average transplanted tissue volume at the end of drug administration and at the beginning of drug administration (d0), respectively, for a certain treatment group; V i V0 and V0 represent the mean tumor volume at the end of treatment and at the start of treatment (d0) in the solvent control group, respectively. At the end of the trial, the weight of the transplanted tissue was measured. The relative weight of the transplanted tissue reflects the proliferation rate relative to the transplanted tissue (weight), calculated using the formula: W0 T / W C IR reflects the inhibition rate relative to transplanted tissue (weight), and is calculated using the formula: IR = (W C -W T ) / W C ×100%, of which, W T and W C The average transplanted tissue weights are for the treatment group and the solvent control group, respectively.

[0071] Statistical analysis

[0072] All in vivo data are expressed as the standard error of the mean (SEM). Transplanted tissue volume, transplanted tissue weight, and animal body weight were used for statistical analysis to calculate the mean transplanted tissue volume, mean transplanted tissue weight, and SEM for each group at different time points. Examples 1 and 3 were plotted using Graphpad Prism 8.0 software, and data analysis was performed using SPSS 19.0. One-way ANOVA was used to check for homogeneity of variance. If the variances were homogeneous (p>0.05), a one-way ANOVA test was used, and the significance of each group was checked using the LSD test. If the variances were unequal (p<0.05), a nonparametric NPAR test was used for pairwise comparisons, and the significance of each group was checked using the Mann-Whitney test to evaluate the differences in tumor volume and tumor weight between the drug treatment groups and the solvent control group at the end of the efficacy experiment. For all statistical assessments, the significance level is set at p<0.05 to report the significance compared to the control group; that is, p<0.05 is considered statistically significant, and p<0.01 indicates a highly significant statistical difference.

[0073] In Examples 1 and 3, the experimental animals were BALB / c nude and NSG female mice, 6-8 weeks old, weighing 18-22g, purchased from Jiangsu Jicui Pharmaceutical Biotechnology Co., Ltd. and Shanghai Southern Model Biotechnology Co., Ltd., respectively. The test animals underwent acclimatization rearing at the experimental site for 3-7 days prior to the experiment. CYH33 in these examples was provided by Shanghai Haihe Pharmaceutical Research and Development Co., Ltd., and BYL719 was a commercially available product.

[0074] Example 1

[0075] In vivo pharmacodynamic study of CYH33 on the subcutaneous allogeneic transplantation model of the stable cell line NIH / 3T3-PIK3CA H1047R Inquiry

[0076] In this experiment, the stable transgenic cell line NIH / 3T3-PIK3CA H1047R was used to evaluate the anti-tissue growth efficacy of CYH33. NIH / 3T3 cells are a highly contact-inhibited continuous cell line established from NIH Swiss mouse embryo cultures, retaining some normal growth characteristics. They do not form clones in soft agar and cannot proliferate in immunodeficient mice. These cells are easily transfected and are commonly used in cell, molecular biology, and tissue transplantation. Based on the human PIK3CA gene sequence (NM_006218.4), the PIK3CA-H1047R (CAT>CGA) mutant gene fragment was designed and obtained. The PIK3CA-H1047R gene was transferred into wild-type cells NIH / 3T3 (ATCC, Cat#:CRL-1658) using a retroviral packaging system. After screening with puromycin antibiotic, a stable transgenic cell line NIH / 3T3-PIK3CA H1047R was obtained. The constructed stable transgenic NIH / 3T3-PIK3CA H1047R cells were then introduced subcutaneously into nude mice to form overgrown tissue, which was used to screen and evaluate the efficacy of the test compounds.

[0077] Experimental methods

[0078] The NIH / 3T3-PIK3CA H1047R cell line was cultured in 1640 medium with 10% fetal bovine serum and 1% penicillin / streptomycin antibiotics at 37°C in a 5% CO2 incubator, passaged twice a week. When the cell saturation reached 80%–90% and the desired number was achieved, cells in the logarithmic growth phase were harvested, centrifuged at 1000 rpm for 5 min to remove the supernatant, resuspended in culture medium, and counted using a cell counter. Based on the counting results, the original solution was diluted to a viable cell concentration of 1×10⁻⁶. 7Cell suspensions with a cell viability of 98.6% and passage P5 were prepared by diluting the cell suspension with matrix gel at a 1:1 ratio. The mixtures were then placed on ice. Under aseptic conditions, 0.2 mL of the cell suspension was subcutaneously injected into the right axilla of each mouse, equivalent to 1 × 10n NIH / 3T3-PIK3CA H1047R cells per mouse. 6 Observe the occurrence of transplanted tissue. When the average volume of transplanted tissue reaches 132.5 mm², 3 Mice were randomly divided into 3 groups (n=5 per group) and treated with solvent control, CYH33 (7.5 mg / kg and 11.25 mg / kg), respectively. The mice were administered the drug twice a day for 16 consecutive days according to the study dosage regimen shown in Table 1. The efficacy was studied.

[0079] Table 1. Pharmacodynamic groupings and dosing regimens

[0080]

[0081] Note: a. BID: Administer twice daily;

[0082] b.PO: Oral administration;

[0083] The solvent for c.CYH33 is: 0.5% sodium carboxymethyl cellulose (CMC-Na) + 0.5% Tween 80.

[0084] Experimental results

[0085] The inhibitory effect of CYH33 on tissue growth in female nude mice bearing subcutaneous xenografts of NIH / 3T3-PIK3CA H1047R cells is as follows: Figure 1-2 Tables 2-3 show the grouping and dosing regimens for each group, as shown in Table 1. Among them, Figure 1 The growth curves of transplanted tissues in each treatment group of the CYH33 model in the NIH / 3T3-PIK3CA H1047R allogeneic transplantation model are shown. Figure 2 The effects of each treatment group on body weight changes in mice with a subcutaneous allogeneic transplantation model of NIH / 3T3-PIK3CA H1047R cells were shown. Figure 1 , 2 The data points represent the average transplanted tissue volume and average weight change rate for each group, respectively, and the error bars represent the standard error (SEM) of the mean. Tables 2 and 3 show the average transplanted tissue volume, average transplanted tissue weight, and the transplanted tissue response indicators T / C, TGI, and IR (%) for each treatment group after 16 days of drug administration. In principle, the evaluation criteria were: T / C (%) > 40% was considered ineffective; T / C (%) ≤ 40% and p < 0.05 was considered effective.

[0086] Table 2. Effects of CYH33 on tissue growth in the NIH / 3T3-PIK3CA H1047R cell allogeneic transplantation model (calculated based on transplanted tissue volume on day 16 after drug administration).

[0087]

[0088] Note: a.Mean±SEM;

[0089] bT / C(%)=T RTV / C RTV ×100, RTV=V 16 / V0;TGI(%)=[1-(T 16 -T0) / (C 16 -C0)]×100;

[0090] c. The differences between the CYH33 dosage groups and the solvent control group were obtained using the nonparametric Mann-Whitney test.

[0091] Table 3. Transplanted tissue weights in each group (based on day 16 after drug administration)

[0092]

[0093] Note: a.Mean±SEM;

[0094] b. IR (%) = (W C -W T ) / W C ×100%;

[0095] c. The differences between the CYH33 dosage groups and the solvent control group were obtained using the nonparametric Mann-Whitney test.

[0096] Depend on Figure 1 As shown in Table 2, 16 days after the start of drug administration, the average transplanted tissue volume of mice in the solvent control group was 2072±83 mm. 3 Compared with the blank solvent group, CYH33 significantly inhibited tissue growth at doses of 7.5 and 11.25 mg / kg, with mean tissue volumes of 740 ± 163 mm². 3 (T / C = 35%, TGI = 69%, p = 0.009) and 474 ± ​​57 mm 3(T / C = 23%, TGI = 82%, p = 0.009). Furthermore, the effect of each treatment group on tissue weight was investigated in the experiment. After 16 consecutive days of administration, the transplanted tissues of mice in each treatment group were weighed. As shown in Table 3, the anti-tissue growth effects of each treatment group were significantly different from those of the solvent control group. The average tissue weight of the control group mice was 2.89 ± 0.36 g. After treatment with CYH33 (7.5, 11.25 mg / kg), the average tissue weights were 0.90 ± 0.18 g (IR = 69%, p = 0.009) and 0.75 ± 0.13 g (IR = 74%, p = 0.009), respectively. All animals tolerated the treatment well.

[0097] In summary, the test compound CYH33 demonstrated a significant and effective anti-transplant tissue growth effect in the NIH / 3T3-PIK3CA H1047R subcutaneous transplantation model, and all animals tolerated it well. These data provide a rationale for the clinical application of CYH33 in the treatment of patients with PIK3CA-associated excessive growth syndrome (PROS) and / or PIK3CA-associated vascular malformations (PRVM).

[0098] Example 2

[0099] Effects of CYH33 on the in vitro proliferation of HMEC-1-PIK3CA-H1047R cells

[0100] In this experiment, the in vitro efficacy of CYH33 was evaluated using a constructed stable HMEC-1 cell line overexpressing the target gene PIK3CA-H1047R (CAT>CGA) mutation. HMEC-1 is a continuously regenerating human endothelial microvascular cell that retains many characteristics of endothelial cells and can be used as a substitute for primary human dermal endothelial cells in studies such as angiogenesis. The PIK3CA-H1047R (CAT>CGA) mutant target gene fragment was designed and obtained based on the human PIK3CA gene sequence (NM_006218.4). The PIK3CA-H1047R gene was transformed into HMEC-1 cells (Zhong Qiao Xinzhou, #ZQ0456) using a retroviral packaging system. After screening with puromycin antibiotic, a stable HMEC-1-PIK3CA-H1047R transgenic cell line was obtained.

[0101] Experimental methods

[0102] Cells were stained with trypan blue and viable cells were counted. The cell concentration was adjusted to an appropriate level and seeded into 96-well plates. Cell-free culture medium (containing 0.1% DMSO) was added to the min control wells. The plates were incubated overnight at 37°C. The maximum final concentration of the test compounds CYH33 and BYL719 was 10 μM, with nine concentrations set from 10 μM to the minimum. The final DMSO concentration was 0.1%. The plates were incubated at 37°C in a 5% CO2 incubator. After 7 days, cell viability was assessed using the CellTiter-Glo luminescence assay kit (Promega-G7573).

[0103] Experimental results

[0104] As shown in Table 4, Figure 3 As shown, CYH33 inhibits the proliferation of HMEC-1-PIK3CA-H1047R cells, with a relative IC50 value of 1 / 3. 50 The IC50 value was 187 nM, while the relative IC50 value of the drug BYL719 targeting the same target was 187 nM. 50 The value was 1167 nM. Therefore, CYH33's ability to inhibit the proliferation of PIK3CA-mutated angiocytes was significantly stronger than that of BYL719.

[0105] Table 4. Anti-proliferation parameters of CYH33 in HMEC-1-PIK3CA H1047R cells

[0106]

[0107] Example 3

[0108] In vivo pharmacodynamic study of CYH33 on the subcutaneous allogeneic transplantation model of the stable cell line NIH / 3T3-FL-TIE2-L914F Inquiry

[0109] In this experiment, the stable transgenic cell line NIH / 3T3-FL-TIE2-L914F was used to evaluate the anti-tissue growth efficacy of CYH33. NIH / 3T3 cells are a highly contact-inhibited continuous cell line established from NIH Swiss mouse embryo cultures, retaining some normal growth characteristics. They do not form clones on soft agar and cannot proliferate in immunodeficient mice. These cells are easily transfected and are commonly used in cell, molecular biology, and tissue transplantation. The FL-TIE2-L914F (CTT>TTT) mutant expression vector was designed and obtained based on the FL-TIE2 sequence (NM_000459.5). The FL-TIE2-L914F gene was transformed into wild-type cells NIH / 3T3 (ATCC, Cat#:CRL-1658) using a retroviral packaging system. After screening with puromycin antibiotic, a stable transgenic cell line NIH / 3T3-FL-TIE2-L914F was obtained. The constructed stable transgenic NIH / 3T3-FL-TIE2-L914F cells were then introduced into nude mice to form overgrown tissue, which was used to screen and evaluate the efficacy of the experimental compounds.

[0110] Experimental methods

[0111] The NIH / 3T3-FL-TIE2-L914F cell line was cultured in DMEM medium supplemented with 10% fetal bovine serum and 1% penicillin / streptomycin antibiotics at 37°C in a 5% CO2 incubator, and passaged twice a week. When the cell saturation reached 80%–90% and the required number was achieved, cells in the logarithmic growth phase were harvested. Under aseptic conditions, 0.2 mL of cell suspension was subcutaneously inoculated into the right axilla of each mouse, i.e., 1 × 10n NIH / 3T3-FL-TIE2-L914F cells per mouse. 6 Observe the occurrence of transplanted tissue. When the average volume of transplanted tissue reaches 102 mm², observe the development of transplanted tissue. 3 At that time, tumor-bearing mice were randomly divided into 3 groups (n=5 per group), and were treated with solvent control and CYH33 or BYL719 respectively. The drugs were administered according to the study dosage regimen shown in Table 5, twice a day for 14 consecutive days, to study the efficacy.

[0112] Table 5. Pharmacodynamic groupings and dosing regimens Note: a. BID: Administer twice daily;

[0113] b.PO: Oral administration;

[0114] The solvent for c.BYL719 is: 10% N-methylpyrrolidone + 30% PEG300 + 20% Solutol + 40% ultrapure water;

[0115] d. From day 0 to day 10 after administration, the dose of CYH33 was 7.5 mg / kg; from day 11 to day 14 after administration, the dose was 11.25 mg / kg, and the corresponding concentration was adjusted to 1.125 mg / mL.

[0116] The solvent and solvent control for e.CYH33 are: 0.5% CMC-Na + 0.5% Tween 80.

[0117] Experimental results

[0118] The inhibitory effect of CYH33 on tissue growth in female nude mice bearing NIH / 3T3-FL-TIE2-L914F cell subcutaneous xenograft tissue is as follows: Figure 4-5 As shown in Table 6-7. Among them, Figure 4 The growth curves of transplanted tissues in different treatment groups are shown in a nude mouse model bearing NIH / 3T3-FL-TIE2-L914F cell subcutaneous xenograft tumors. Figure 5 The effects of each treatment group on the change in body weight in mice with a subcutaneous allogeneic cell transplantation model were shown. Figure 4 , 5 The data points represent the mean transplanted tissue volume and mean weight change rate for each group, respectively, and the error bars represent the standard error (SEM) of the mean. Tables 6 and 7 show the mean transplanted tissue volume, mean transplanted tissue weight, and the indicators of transplanted tissue response to treatment (T / C, TGI, and IR) for each treatment group 14 days after administration. In principle, the evaluation criteria were: T / C (%) > 40% was considered ineffective; T / C (%) ≤ 40% and p < 0.05 was considered effective.

[0119] Table 6. Efficacy evaluation of CYH33 on tissue growth in NIH / 3T3-FL-TIE2-L914F cell allograft model

[0120] (Calculated based on the transplanted tissue volume on day 14 after drug administration)

[0121]

[0122] Note: a.Mean±SEM;

[0123] bT / C(%)=T RTV / C RTV ×100, RTV=V 14 / V0;TGI(%)=[1-(T 14 -T0) / (C 14 -C0)]×100;

[0124] c. The difference between the drug administration group and the solvent control group was obtained using the nonparametric Mann-Whitney test;

[0125] d. The dose of CYH33 was 7.5 mg / kg on days 0 to 10 after administration, and 11.25 mg / kg on days 11 to 14 after administration.

[0126] Table 7. Transplanted tissue weights in each group (based on day 14 after drug administration)

[0127]

[0128] Note: a.Mean±SEM;

[0129] b. IR (%) = (W C -W T ) / W C ×100%;

[0130] c. One-way ANOVA was used to analyze the differences between the drug administration group and the solvent control group;

[0131] d. The dose of CYH33 was 7.5 mg / kg on days 0 to 10 after administration, and 11.25 mg / kg on days 11 to 14 after administration.

[0132] Depend on Figure 4 As shown in Table 6, 14 days after the start of drug administration, the average transplanted tissue volume of mice in the solvent control group was 2486±230 mm. 3 Compared with the blank solvent group, the CYH33 treatment group significantly inhibited tissue growth, with an average transplanted tissue volume of 982±70 mm. 3 (T / C = 40%, TGI = 63%, p = 0.009), and stronger than the BYL719 treatment group (1703 ± 201 mm). 3 The T / C ratio was 68%, TGI was 33%, and p was 0.076. Furthermore, the effect of each treatment group on tissue weight was investigated in the experiment. After 14 days of continuous administration, the transplanted tissues of mice in each treatment group were weighed. Table 7 shows that the average tissue weight of the control group mice was 3.26 ± 0.26 g. The anti-tissue growth effect of the CYH33 treatment group was significantly different from that of the solvent control group (average tissue weight 1.16 ± 0.11 g, IR = 64%, p = 0.009), and stronger than that of the BYL719 treatment group (average tissue weight 1.74 ± 0.21 g, IR = 47%, p = 0.009). All animals tolerated the treatment well.

[0133] In summary, the test compound CYH33 demonstrated a significant and effective inhibitory effect on transplanted tissue growth in the NIH / 3T3-FL-TIE2-L914F subcutaneous transplantation model, and its effect was stronger than that of the BYL719 treatment group. These data provide a rationale for the clinical application of CYH33 in treating patients with TEK gene mutation-related venous malformations.

[0134] Example 4

[0135] Effects of CYH33 on the in vitro proliferation of HMEC-1-TIE2-L914F cells

[0136] In this experiment, the FL-TIE2-L914F (CTT>TTT) mutant expression vector was designed and obtained based on the FL-TIE2 sequence (NM_000459.5). The TIE2-L914F gene was transformed into HMEC-1 cells (Zhong Qiao Xinzhou, #ZQ0456) using a lentiviral packaging system. After screening with puromycin antibiotic, a stable transfected cell line, HMEC-1-TIE2-L914F, was obtained. The stable HMEC-1 cell line overexpressing the target gene TIE2-L914F mutant was used to evaluate the in vitro efficacy of CYH33.

[0137] Experimental methods

[0138] Adjust the cell concentration to an appropriate level, adding 2000 cells / 100 μL of cell suspension to each well of the cell plate. Add cell-free culture medium (containing 0.1% DMSO) to the Min control wells. Incubate the plate overnight at 37°C. The maximum final concentration of the test compounds CYH33 and BYL719 is 10 μM. Prepare 10× working solutions of the compounds using PBS, and serially dilute the test compounds from the highest concentration to the lowest concentration using PBS solution containing 1% DMSO, resulting in 9 concentrations. Remove the cell plate from the previous day and add 11 μL of 10× working solution of the compounds to each well. Add 11 μL of 1% DMSO PBS solution to the Max control well, bringing the final DMSO concentration to 0.1%. Incubate the plate at 37°C in a 5% CO2 incubator. After 7 days, cell viability is assessed using the CellTiter-Meiluncell luminescence assay kit (PWL111-3).

[0139] Experimental results

[0140] As shown in Table 8, Figure 6 As shown, CYH33 inhibits the proliferation of HMEC-1-TIE2-L914F cells, relative to IC50. 50 The concentration was 291.8 nM, and its ability to inhibit the proliferation of TIE2-mutant angiocytes was stronger than that of the drug BYL719 with the same target (relative IC50). 50 (3179 nM).

[0141] Table 8. Anti-proliferation parameters of CYH33 in HMEC-1-TIE2-L914F cells

[0142]

[0143] Example 5

[0144] Effect of CYH33 on p-AKT expression in HMEC-1-TIE2-L914F cells

[0145] Experimental methods

[0146] HMEC-1-TIE2-L914F cells were cultured in HMEC-1 complete medium (Zhongqiao Xinzhou, #ZQ-1319). An appropriate number of cells were seeded into 6-well plates and allowed to adhere overnight. The next day, the compound was added to the seeded cell culture plates at the specified concentration, with DMSO used as a control well. Incubation was performed for 2 hours. Cell pellets were lysed using RIPA lysis buffer, which included protease and phosphatase inhibitors. After lysis, the protein concentration of each sample was determined using a BCA protein assay kit, and the protein concentration was uniformly quantified. 5× protein loading buffer was added, and the samples were boiled at 95°C for 10 min, cooled on ice, and stored at -20°C. Cell protein samples were added to the sample wells of the gel and loaded onto the Protein Ladder. Electrophoresis was performed at a constant voltage of 70V for 30 min, followed by a constant voltage of 86V until the bromophenol blue indicator band reached the bottom. Electrophoresis was then stopped. The proteins on the SDS gel were transferred to a PVDF membrane at a constant voltage of 100V for 105 min. After electroporation, the PVDF membrane was placed in blocking buffer (5% skim milk) and blocked at room temperature for 1 hour. The blocked PVDF membrane was washed, cut, and incubated overnight at 4°C with primary antibodies (1:1000 dilution) p-AKT (Ser473) (CST, #4060S), AKT (CST, #4691S), and GAPDH (Trans, #HC301), respectively. The overnight PVDF membrane was slowly washed three times in TBST and then incubated at room temperature for 1 hour with the corresponding secondary antibodies (1:3000 dilution) HRP-anti-mouse IgG (CST, #7076S) and HRP-anti-Rabbit IgG (CST, #7074S), respectively. After secondary antibody incubation, the PVDF membrane was slowly washed three times, and the membrane was exposed under a chemiluminescence imaging system to save the images. Figure 7 a). Image J2x software was used to perform grayscale analysis on the p-AKT and AKT bands in the Western Blot results. The grayscale results are presented in bar charts. Figure 7b), where the horizontal axis of the bar chart represents the compound concentration, and the vertical axis represents the ratio of p-AKT to AKT (p-AKT / AKT), with the formula: p-AKT / AKT (% control) = Gray compound / Gray DMSO × 100%.

[0147] Experimental results

[0148] like Figure 7 As shown, in HMEC-1-TIE2-L914F cells, the tested compound CYH33 exhibited a strong dose-dependent inhibitory effect on p-AKT (Ser473). Specifically, at concentrations of 100 nM and 300 nM, CYH33 significantly inhibited the expression of AKT phosphorylation.

[0149] Example 6

[0150] Clinical trials of CYH33

[0151] A phase I / II, multicenter clinical study was conducted to clinically evaluate the safety, tolerability, pharmacokinetic characteristics, and efficacy of CYH33 in adult and adolescent (12–17 years) patients with severe clinical presentations, requiring systemic therapy, and who have a confirmed PIK3CA hotspot mutation in PROS and PRVM.

[0152] Specifically, a Phase I clinical trial was conducted in adult patients with PROS / PRVM. CYH33 was administered orally once daily (QD) for 21 days as one treatment cycle (the first treatment cycle in the Phase I study was 28 days), starting with a dose of 10 mg QD. Seventeen adult patients with PROS / PRVM received CYH33 treatment (5, 10, or 15 mg, QD). In the Phase I dose escalation phase, 4, 3, and 3 patients were enrolled at the 5 mg QD, 10 mg QD, and 15 mg QD dose levels, respectively. In the Phase I dose extension phase, 7 patients were enrolled at the 10 mg QD dose level. Patients were aged 18–46 years and weighed 45.8–108.4 kg; 5 were PROS patients and 12 were PRVM patients. PK samples were obtained from 16 adult patients with PROS / PRVM during the dose escalation and expansion phases, with 3, 10, and 3 patients receiving 5 mg, 10 mg QD, and 15 mg QD, respectively. The aim was to evaluate the safety, tolerability, pharmacokinetic characteristics, and preliminary efficacy of CYH33 administration in PROS / PRVM patients.

[0153] Regarding clinical safety, the most common drug-related adverse events in PROS / PRVM patients treated with CYH33 monotherapy at 5-15 mg QD in this study were hyperglycemia and stomatitis. Compared to the higher dose levels (20-60 mg QD) used in solid tumor patients, the incidence and severity of adverse events were significantly reduced, and all were CTCAE grade 1-2. The most common drug-related adverse events were CTCAE grade 1 hyperglycemia and CTCAE grade 1 or 2 stomatitis (see Table 9 for details), and no DLT events occurred in any patient. Furthermore, no adverse events such as diarrhea or loss of appetite have been reported to date.

[0154] Table 9. Preliminary safety data of CYH33 in adult patients with PROS / PRVM (≥10% TRAE)

[0155]

[0156]

[0157] Regarding clinical efficacy, all 16 participants in Phase I completed at least one efficacy assessment. In the CYH33 5mg QD group, 2 out of 3 participants achieved partial remission (PR) at the first imaging assessment; in the CYH33 10mg QD group, 9 out of 10 participants achieved PR at the first imaging assessment; and in the CYH33 15mg QD group, all 3 participants achieved PR at the first imaging assessment. Please refer to [link to details]. Figure 8 .

[0158] Besides the reduction in target lesion size, improvements in patients' clinical symptoms and quality of life are also important indicators of treatment efficacy, as shown in Table 10. The most common clinical symptoms in PROS / PRVM patients are overgrowth / malformation, pain, and fatigue. Preliminary data show that with CYH33 5-15mg QD treatment, the improvement rate for pain was 75-100%, the improvement rate for fatigue was 50.0-100%, the improvement rate for overgrowth / malformation was 90-100%, and the improvement rate for quality of life score was 25-67%.

[0159] Table 10. Improvement in clinical symptoms and quality of life in adult patients with PROS / PRVM after CYH33 treatment

[0160]

[0161] In summary, CYH33 is a highly selective PI3Kα inhibitor. Preliminary data from a Phase I clinical trial in adult patients with PROS / PRVM show that CYH33 exhibits good safety and pharmacokinetic characteristics at dose levels of 5-15 mg QD, and demonstrates significant efficacy, reducing lesions by 20% and even 30-65%. These results provide data support for the treatment of PROS / PRVM patients with CYH33.

Claims

1. Use of CYH33 or a pharmaceutical composition containing CYH33 in the preparation of a medicament for the prevention, treatment or relief of PIK3CA-related malformations, especially PIK3CA-related overgrowth syndrome and / or PIK3CA-related vascular malformations, and / or TEK gene mutation-related vascular malformations.

2. Use of CYH33 or a pharmaceutical composition containing CYH33 in the preparation of a medicament for the prevention, treatment or relief of PIK3CA-associated overgrowth syndrome, PIK3CA-associated vascular malformations, or TEK gene mutation-associated vascular malformations.

3. Use of CYH33 or a pharmaceutical composition containing CYH33 in the preparation of a medicament for the prevention, treatment or relief of PIK3CA-associated hyperglycemia and PIK3CA-associated vascular malformations.

4. Use of CYH33 or pharmaceutical compositions containing CYH33 in the preparation of medicaments for the prevention, treatment or relief of PIK3CA-related malformations and TEK gene mutation-related vascular malformations.

5. The use according to any one of claims 1-4, wherein, The vascular malformation mentioned refers to either a venous malformation or a lymphatic malformation.

6. The use according to any one of claims 1-4, wherein, The pharmaceutical composition comprises a therapeutically effective amount of CYH33, optionally one or more other therapeutic agents, and pharmaceutically acceptable excipients. Preferably, the pharmaceutical composition is formulated for intravenous, intramuscular, oral, rectal, inhalation, nasal, topical, ocular, or ocular administration.

7. The use according to any one of claims 1-4, wherein, The pharmaceutical composition is a tablet, pill, capsule, liquid, inhaler, nasal spray solution, suppository, solution, emulsion, ointment, eye drop, or ear drop.

8. The use according to any one of claims 1-3, wherein, The PIK3CA-related overgrowth syndromes include: macrodactyly, Cloves syndrome, Clappo syndrome, fibrofatty hyperplasia or overgrowth, Klippel-Trenaunay syndrome, unilateral hyperplastic lipomatosis, macrocephaly-capillary malformation, diffuse capillary malformation with overgrowth, fibrofatty vascular lesions, facial invasive lipoma, dysplastic macrocephaly, hemilateral macrocephaly, unilateral muscle overgrowth, seborrheic keratosis, and benign lichenoid keratosis. Preferably, the PIK3CA-related overgrowth syndrome is Cloves syndrome, Klippel-Trenaunay syndrome, fibrofatty hyperplasia or overgrowth, unilateral hyperplastic lipomatosis, or macrocephaly-capillary malformation. The PIK3CA-related vascular malformations include: simple vascular malformations (such as capillary malformations, venous malformations, and lymphatic malformations) and mixed vascular malformations.

9. The use according to claim 8, wherein, The lymphatic malformations include: common lymphatic malformations (such as giant cystic lymphatic malformations, microcystic lymphatic malformations, and mixed cystic lymphatic malformations), generalized lymphatic abnormalities, lymphatic malformations in Gorham-Stout syndrome, tubular lymphatic malformations, and idiopathic lymphedema. The capillary malformations include: simple vascular nevi / salmon spots, skin and / or mucous membrane capillary malformations, reticular capillary malformations, capillary malformations in arteriovenous malformations, congenital telangiectasia with marble-like skin, and telangiectasia. The venous malformations include common venous malformations.

10. The use according to claim 1, 2 or 4, wherein, The TEK gene mutation-related vascular malformations (especially venous malformations) include: common venous malformations, familial mucocutaneous venous malformations, and blue rubber nipple syndrome.

Images