Use of compound in treatment of PIK3ca-related overgrowth spectrum

Abstract

The use of a compound in the treatment of PIK3CA-related overgrowth spectrum. Specifically, disclosed is the use of a compound I, or an isotopic derivative or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating and / or preventing PIK3CA-related overgrowth spectrum.

Description

Use of compounds for treating PROS spectrum diseases

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to Chinese patent application No. 202311108521.4, filed on August 30, 2023. The entire text of this Chinese patent application is incorporated herein by reference. Technical Field

[0003] The present invention relates to the field of biomedicine technology, and in particular to use of a compound for treating PROS spectrum diseases. Background Art

[0004] PROS spectrum disorder (PIK3CA-related overgrowth spectrum) is a rare overgrowth syndrome characterized by congenital lipoma overgrowth, vascular malformations, epidermal nevi, scoliosis / skeletal deformities, or spinal cord abnormalities. PROS spectrum disorders include: facial infiltrating lipomatosis (FIL), fibroadipose hyperplasia or overgrowth (FAO), hemihyperplasia-multiple lipomatosis (HHML), CLOVES syndrome, isolated large lymphatic malformation (ILM), macrodactyly, muscular hemihyperplasia (HH), megalencephaly, CLAPO syndrome, fibroadipose vascular anomaly, Klippel-Trenaunay syndrome, hemimegalencephaly, and megalencephaly-capillary malformation. malformation, MCAP), skin lesions including benign lichenoid keratosis (BLK), epidermal nevus (EN) and seborrheic keratosis (SK). PROS spectrum diseases have a variety of clinical manifestations, ranging from local tissue abnormal proliferation to a wide spectrum of growth malformations of multiple tissues and organs, from mild local and isolated vascular malformations, lipoma or fibroma overgrowth to progressive and extensive lipoma overgrowth associated with life-threatening vascular malformations and related complications (Keppler-Noreuil KM et al., PIK3CA-related overgrowth spectrum (PROS): diagnostic and testing eligibility criteria, differential diagnosis, and evaluation. Am J Med Genet A. 2015 Feb; 167A(2): 287–295).

[0005] PROS can be considered a mixture of highly anatomically diverse overgrowth tissues, with the vasculature (capillaries, veins, and lymphatics) and adipose tissue typically being the most prominently affected macroscopically. Many other tissues and organs, including bone, brain, peripheral nerves, liver, skeletal, and myocardial tissue, may also be affected. Complications of PROS include functional impairment (e.g., walking or swallowing), pain, recurrent superficial infections, thromboembolism, and / or bleeding, all of which can be debilitating and lead to early death (Kurek KC et al., Somatic mosaic activating mutations in PIK3CA cause CLOVES syndrome. Am J Hum Genet. 2012, 90: 1108–1115; Keppler-Noreuil, KM, et al., Clinical delineation and natural history of the PIK3CA-related overgrowth spectrum. Am J Med Genet A, 2014, 164a(7): 1713–1733). In addition to being life-threatening, PROS also causes damage to the patient's appearance and body throughout their lives, which greatly impairs their social function and causes them to bear tremendous psychological pressure.

[0006] PROS spectrum disorders can be alleviated with standard treatments such as vascular sclerotherapy and surgical tumor reduction. However, standard treatments are still incurable and may even lead to recurrence. Secondly, PROS clinical manifestations are heterogeneous and complex, often involving multiple organ systems and various tissue types. The risks and complexity of traditional sclerotherapy and surgical treatments make them difficult to promote, posing a serious challenge to traditional treatment methods (Mirzaa G et al., PIK3CA-Related Overgrowth Spectrum. 2013 Aug 15 [Updated 2021 Dec 23]. In: Adam MP, Ardinger HH, Pagon RA, et al., editors. [Internet]. Seattle (WA): University of Washington, Seattle; 1993–2022; Hardwicke J, Khan MAA, Richards H, Warner RM, Lester R. Macrodactyly—options and outcomes. J Hand Surg Eur Vol. 2013; 38:297–303).

[0007] In addition, alpelisib (BYL719) received FDA approval in April 2022 for the treatment of PROS patients aged 2 years and above. However, existing treatment options for PROS spectrum disorders are still very limited, and the development of new treatments is urgently needed.

[0008] Summary of the Invention

[0009] The present invention aims to provide a compound for treating PROS spectrum diseases.

[0010] The present invention provides a use of Compound I, its isotopic derivatives or pharmaceutically acceptable salts in the preparation of a medicament for treating and / or preventing PROS spectrum diseases;

[0011] The present invention also provides a method for treating and / or preventing a PROS spectrum disease in a patient, comprising administering to the patient an effective amount of Compound I, an isotopic derivative thereof, or a pharmaceutically acceptable salt thereof;

[0012] The present invention also provides a method for inhibiting the growth of PROS lineage disease cells in vitro, in vivo or ex vivo, comprising contacting an effective amount of Compound I, its isotopic derivative or pharmaceutically acceptable salt with the PROS lineage disease cells;

[0013] The present invention also provides a pharmaceutical composition for treating and / or preventing PROS spectrum diseases, comprising Compound I, its isotopic derivative or pharmaceutically acceptable salt;

[0014] In some embodiments, the pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients.

[0015] The present invention also provides a combination kit comprising:

[0016] A kit (a) comprising a detection reagent for detecting a PROS spectrum disorder; and

[0017] A kit (b) comprising a therapeutic agent for treating a PROS spectrum disease, wherein the therapeutic agent comprises Compound 1, an isotopic derivative thereof, or a pharmaceutically acceptable salt thereof as an active ingredient;

[0018] In some embodiments, the detection reagent for detecting PROS spectrum diseases includes a reagent for detecting PIK3CA gene mutation.

[0019] In some embodiments, the PROS spectrum disorder is one or more of infiltrating lipoma of the face, fibrofatty hyperplasia or overgrowth, hemiplasia with multiple lipomatosis, CLOVES syndrome, isolated macrolymphatic malformation, macrodactyly, hemihypertrophy, megalencephaly syndrome, CLAPO syndrome, fibro-fatty vascular anomaly, KT syndrome, hemimegalencephaly, megalencephaly-capillary malformation, benign lichenoid keratosis, epidermal nevus, and seborrheic keratosis.

[0020] In some embodiments, the PROS spectrum disease is a PROS spectrum disease with PIK3CA H1047R mutation. In some embodiments, the PROS spectrum disease with PIK3CA H1047R mutation is one or more of the following PROS spectrum diseases with PIK3CA H1047R mutation: infiltrating lipoma of the face, fibrofatty hyperplasia or overgrowth, hemiplasia with multiple lipomatosis, CLOVES syndrome, isolated macrolymphatic malformation, macrodactyly, hemihypertrophy, megalencephaly syndrome, CLAPO syndrome, fibro-fatty vascular anomaly, KT syndrome, hemi-megalencephaly, megalencephaly-capillary malformation, benign lichenoid keratosis, epidermal nevus, seborrheic keratosis.

[0021] In some embodiments, the PROS spectrum disorder is infiltrating lipoma of the face.

[0022] In some embodiments, the facial infiltrating lipoma is a facial infiltrating lipoma with a PIK3CA H1047R mutation.

[0023] In some embodiments, Compound 1, its isotopic derivative, or pharmaceutically acceptable salt is used as the sole active ingredient for treating and / or preventing PROS spectrum diseases.

[0024] In some embodiments, the Compound 1, its isotopic derivative or pharmaceutically acceptable salt is used in combination with other active ingredients for the treatment and / or prevention of PROS spectrum diseases. The other active ingredients can be drugs known for the treatment and / or prevention of PROS spectrum diseases or drugs that are helpful for the treatment and / or prevention of PROS spectrum diseases.

[0025] Compound 1, its isotopic derivatives or pharmaceutically acceptable salts can be administered in any suitable manner, including oral administration, injection, etc. Depending on the mode of administration, Compound 1, its isotopic derivatives or pharmaceutically acceptable salts can be formulated into various suitable dosage forms, including tablets, etc. In some embodiments, Compound 1, its isotopic derivatives or pharmaceutically acceptable salts can be formulated for oral administration.

[0026] Unless otherwise indicated, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be construed as indefinite or unclear unless specifically defined, but rather should be understood in accordance with its ordinary meaning. When a trade name appears in this document, it is intended to refer to the corresponding commercial product or its active ingredient.

[0027] The term "pharmaceutically acceptable" refers to those compounds, materials, compositions and / or dosage forms which, within the scope of sound medical judgment, are suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0028] The term "pharmaceutically acceptable salts" refers to salts of the compounds of the present invention prepared with relatively non-toxic, pharmaceutically acceptable acids or bases.

[0029] The term "treat" refers to therapeutic treatment. When referring to a specific condition, treatment means: (1) alleviating the disease or one or more biological manifestations of the condition, (2) interfering with (a) one or more points in the biological cascade that leads to or causes the condition or (b) one or more biological manifestations of the condition, (3) ameliorating one or more symptoms, effects, or side effects associated with the condition or one or more symptoms, effects, or side effects associated with the condition or its treatment, or (4) slowing the progression of the condition or one or more biological manifestations of the condition.

[0030] The term "prevent" refers to the reduction of the risk of acquiring or developing a disease or disorder.

[0031] The term "effective amount" refers to an amount of a compound that, when administered to a patient, is sufficient to effectively treat and / or prevent a disease or condition described herein. The "effective amount" will vary depending on the compound, the condition and its severity, and the age of the patient to be treated, but can be adjusted as needed by those skilled in the art.

[0032] The term "pharmaceutically acceptable excipients" refers to excipients and additives used in the production of medicines and the preparation of prescriptions. It is all substances contained in pharmaceutical preparations in addition to the active ingredients.

[0033] The term "patient" refers to any animal to which a drug is to be or has been administered, preferably a mammal, and most preferably a human. The term "mammal" includes any mammal. Examples of mammals include, but are not limited to, cows, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, monkeys, and humans, with humans being the most preferred.

[0034] The term "isotopic derivative" refers to a derivative formed when an atom in a compound is replaced by an isotope of the atom. Examples of isotopes that can be incorporated into the compounds of the present invention include, but are not limited to, isotopes of hydrogen, carbon, nitrogen, oxygen, sulfur (e.g. 2 H. 3 H. 13 C. 14 C. 15 N. 18 O. 17 O. 35 S). Isotopic derivatives are prepared by substituting an isotopically labeled reagent for a non-isotopically labeled reagent.

[0035] Without violating the common sense in the art, the above-mentioned preferred conditions can be arbitrarily combined to obtain preferred embodiments of the present invention.

[0036] The reagents and raw materials used in the present invention are commercially available.

[0037] The positive and progressive effect of the present invention is that it provides a compound for treating PROS spectrum diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] FIG1 shows the survival curves of adipose-derived mesenchymal stem cells derived from patients with facial infiltrating lipoma after treatment with different concentrations of Compound I.

[0039] FIG2 shows the survival curves of adipose-derived mesenchymal stem cells derived from patients with facial infiltrating lipoma after treatment with different concentrations of BYL719. DETAILED DESCRIPTION

[0040] The present invention is further illustrated by way of examples below, but the present invention is not limited to the scope of the examples. Experimental methods in the following examples where specific conditions are not specified were performed according to conventional methods and conditions, or selected according to the product specifications.

[0041] The compound I in the following examples refers to

[0042] Example 1: Evaluation of the efficacy of Compound I in patient-derived facial infiltrating lipoma cells

[0043] Experimental purpose: To evaluate the in vitro inhibitory effect of compound I on adipose-derived mesenchymal stem cells derived from patients with facial infiltrating lipoma.

[0044] Experimental Materials:

[0045] 1) Fresh facial infiltrating lipoma patient samples (surgical origin, NGS genetic testing showed PIK3CA H1047R mutation, PROS spectrum disease. The patient was from the Ninth People's Hospital affiliated to Shanghai Jiao Tong University School of Medicine).

[0046] 2) Cell Extraction: Cut whole adipose tissue into 0.1–0.2 cm sections. Mix the excised fat (or liposuctioned fat) in equal proportions with serum-free culture medium containing 0.2% collagenase I and digest on a shaker (150 rpm, 37°C) for 1 hour. Collect cells by centrifugation at 1000 rpm for 5 minutes. Remove red blood cells with red blood cell lysis buffer, and filter the cell pellet through a 100 μm filter to collect the cells. Cells were cultured and passaged in a 37°C incubator with 5% CO2. Cells from passages 3–5 were used in this experiment.

[0047] 3) Experimental reagents and instruments:

[0048] Collagenase I (Worthington, USA, LS004196);

[0049] serum-free medium (Cyagen, USA, HUXMD-90062);

[0050] red blood cell lysis buffer (Solarbio, China, R1010);

[0051] Human adipose-derived stem cell growth medium (also referred to as basal medium in this patent, Cyagen, USA, HUXMD-90011);

[0052] CCK-8 kit (Epizyme, China, CX001M);

[0053] Microplate reader (Thermo Fisher, A51119500C).

[0054] Experimental methods:

[0055] Seed plate: 3×10 3 Cells were seeded into 96-well plates at a density of 100 μL of basal medium per well. After 24 hours, the medium was changed after the cells were fully attached.

[0056] Drug administration: Compound I was diluted three-fold in a series of eight dilutions starting at 3 μM, with three replicates per dilution. BYL719 was diluted three-fold in a series of eight dilutions starting at 30 μM, with three replicates per dilution. Each well contained 150 μL of drug-containing medium. CCK-8 assays were performed 72 hours later.

[0057] CCK-8 activity assay: Cell viability was determined using a CCK-8 assay kit. At the designated time points, 10 μL of CCK-8 was added to each well and incubated for 3 hours. OD values ​​were measured at a wavelength of 450 nM.

[0058] Cell viability (%) = (OD 加药 / OD 对照 )×100%

[0059] Inhibition rate (%) = 1-cell survival rate (%)

[0060] Experimental results:

[0061] 1) IC of Compound I and BYL719 for adipose-derived mesenchymal stem cells derived from patients with facial infiltrating lipoma 50 value

[0062] 2) Average survival rate of adipose-derived mesenchymal stem cells derived from patients with facial infiltrating lipoma after treatment with different concentrations of Compound I and BYL719;

[0063] 3) The survival curves of adipose-derived mesenchymal stem cells derived from patients with facial infiltrating lipoma after treatment with different concentrations of Compound I and BYL719 are shown in Figures 1 and 2 .

[0064] Experimental results:

[0065] This study explored the effects of different concentrations of Compound I and BYL719 on the cell activity of adipose-derived mesenchymal stem cells derived from patients with facial infiltrating lipoma. Compound I had a significant inhibitory effect on adipose-derived mesenchymal stem cells derived from patients with facial infiltrating lipoma in a dose-dependent manner. The half-maximal inhibitory concentration (IC 50 ) is 0.36μM. BYL719 also has a significant inhibitory effect on adipose-derived mesenchymal stem cells derived from patients with facial infiltrating lipoma, also showing a dose-dependent effect. BYL719's IC 50 The IC value of compound I is 4.3 μM. 50 Much lower than BYL719.

[0066] Conclusion: Compound I has a significant inhibitory effect on adipose-derived mesenchymal stem cells (ADMSCs) derived from patients with infiltrating facial lipomas. Compound I exhibits superior inhibitory activity against ADMSCs compared to BYL719. Compound I has potential for the treatment of PROS-related disorders.

[0067] Example 2: Evaluation of the efficacy of compounds in a patient-derived lipoma xenograft model

[0068] Objective: To evaluate the tumor inhibitory effect of the compound in a patient-derived lipoma xenograft mouse model.

[0069] Experimental Materials:

[0070] 1) Lipoma samples: Cell processing and culture methods were the same as in Example 1;

[0071] 2) Experimental reagents and instruments: Matrigel (Corning).

[0072] Experimental methods:

[0073] Experimental animals: BALB / C nude mice (purchased from Shanghai Slack, 6 weeks old, female).

[0074] Transplant tumor model construction: 1×10 6 Adipose-derived mesenchymal stem cells from patients with facial infiltrating lipoma were thoroughly mixed with Matrigel and inoculated into the left / right cheek of each mouse. One day after cell transplantation, the animals were randomly divided into two groups, with six mice in each group, for a total of two groups (control group and compound I treatment group). For the mice in the treatment group, they were gavaged once a day at a dose of 0.1 mg / kg of compound I (solvent: 1% methylcellulose and 1% DMSO (dimethyl sulfoxide)). For the mice in the control group, 200 μL of ddH2O was given by gavage daily. After 4 weeks of treatment, the volume and weight of the transplanted tumors of the mice were measured.

[0075] The formula for calculating tumor volume is: V = 0.5a × b 2 , a and b represent the long diameter and short diameter of the tumor, respectively.

[0076] Experimental results:

[0077] Tumor volume and weight at the end of dosing (day 28);

[0078] This study investigated the in vivo anti-tumor efficacy of compound I in a mouse xenograft model derived from a patient with facial infiltrating lipoma. At the end of the experiment, the average tumor volume and weight of the control group were 17.37 mm 3 Compared with the control group, the tumor volume and weight of mice in the compound I group were significantly reduced, which were 3.21 mm 3 and 2.28mg.

[0079] Conclusion: Compound I exhibits significant anti-tumor activity in a mouse model of facial infiltrating lipoma derived from a patient. Compound I is expected to be used to treat PROS-related diseases.

Claims

1. Use of a compound I, an isotopic derivative or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating and / or preventing PROS spectrum diseases; 2. The use according to claim 1, characterized in that The PROS spectrum diseases are one or more of facial infiltrating lipoma, fibrofatty hyperplasia or overgrowth, hemiplasia with multiple lipomatosis, CLOVES syndrome, isolated macrolymphatic malformation, macrodactyly, hemihypertrophy, megalencephaly syndrome, CLAPO syndrome, isolated macrolymphatic malformation, fibro-fatty vascular anomaly, KT syndrome, hemi-megalencephaly, megalencephaly-capillary malformation, benign lichenoid keratosis, epidermal nevus and seborrheic keratosis.

3. The use according to claim 1, characterized in that The PROS spectrum disease is a PROS spectrum disease with PIK3CA H1047R mutation.

4. The use according to claim 1, characterized in that The PROS spectrum disorder is infiltrating lipoma of the face.

5. The use according to claim 4, characterized in that The facial infiltrating lipoma is a facial infiltrating lipoma with PIK3CA H1047R mutation.

6. The use according to any one of claims 1 to 5, characterized in that The Compound I, its isotopic derivative or pharmaceutically acceptable salt is formulated for oral administration.

7. The use according to any one of claims 1 to 5, characterized in that The compound I, its isotope derivative or pharmaceutically acceptable salt is used as the sole active ingredient for treating and / or preventing PROS spectrum diseases.

8. The use according to any one of claims 1 to 5, characterized in that The compound I, its isotope derivative or pharmaceutically acceptable salt is used in combination with other active ingredients to treat and / or prevent PROS spectrum diseases.

9. A method for treating and / or preventing a PROS spectrum disease in a patient, the method comprising administering to the patient an effective amount of Compound I, an isotopic derivative thereof, or a pharmaceutically acceptable salt thereof; 10. A method for inhibiting the growth of PROS lineage disease cells in vitro, in vivo or ex vivo, comprising contacting an effective amount of Compound I, an isotopic derivative or a pharmaceutically acceptable salt thereof with the PROS lineage disease cells;

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