KR20220045481A - Pharmaceutical composition comprising Poziotinib for treating cancer having HER2 mutation - Google Patents

Abstract

One aspect provides: a pharmaceutical composition for treating HER2 mutation cancer which contains an effective amount of poziotinib for treating cancer comprising at least one mutation present in kinase domain (KD), mucosal membrane domain (JMD), or transmembrane domain (TMD) of a HER2 receptor; and a method for treating cancer using the same.

Description

TECHNICAL FIELD [0001] Pharmaceutical composition comprising Poziotinib for treating cancer having HER2 mutation

It relates to a pharmaceutical composition for the treatment of cancer comprising positiotinib for treating cancer having a mutation in the HER2 receptor.

Epidermal growth factor receptor (EGFR) is a protein composed of a receptor part and a tyrosine kinase part, and functions to pass through the cell membrane and transmit signals from outside the cell to the inside of the cell. Although EGFR plays an essential role in normal cell regulation through signal transduction into cells, normal signaling into cells by overexpression of EGFR, or generation of mutations, such as activating EGFR mutations characterized by ligand-independent tyrosine kinase activity It is known that regulation is inhibited, and thus cancer cell growth, differentiation, angiogenesis, metastasis, and resistance expression are induced. Human epidermal growth factor receptor 2 (HER2) is one of the EGFR family of transmembrane receptor tyrosine kinases (RTKs) and is involved in the activation of key signaling systems for cell proliferation and survival. It is a representative tumor factor used as one of the criteria for subtype classification of breast cancer patients, and can be activated by binding to ligands of other receptors after dimerization of receptors with EGFR or HER3/4, which are other EGFR families. Amplification of the HER2 gene has been reported in 20-30% of all breast cancer patients and 28% of gastric cancer patients, and in this patient group, the cancer recurrence rate is high and the survival rate is short and the prognosis is poor (Non-Patent Document 1).

As HER2-directed therapy that targets HER2, drugs using trastuzumab, pertuzumab, Trastuzumab emtansine (T-DM1), lapatinib, or neratinib Although the treatment is known and the survival rate of patients has been greatly increased through their clinical application, it has a poor prognosis for cancers that are resistant to these drugs. In addition, currently, HER2 targeted therapeutics are only applied in clinical practice for HER2 gene amplification cancers, and research is needed on treatment methods for cases in which the HER2 signaling system is activated by HER2 mutation.

Recently, a large-scale genomic study suggested the need for research on HER2 targeted therapeutics to treat cancer containing HER2 somatic gene mutations (Non-Patent Document 2). It has been reported that when a mutation in the kinase domain of the HER2 receptor occurs, it induces activation of the HER2 signaling system and low survival rate regardless of HER2 gene amplification in breast cancer (Non-Patent Document 3). In addition, as a result of analyzing the target exome sequencing results in 111,176 tumors, mutations in the HER2 receptor were detected in 3,851 tumors (3.5%), and 40% of the extracellular domain (ECD) and the kinase domain (KD) were each Although it occupies a significant amount, mutations in the transmembrane domain (TMD) and the mucosal membrane domain (JMD) were also detected at rates of 2.8% and 7.7% (Non-Patent Document 4). In addition, when comparing the incidence of mutations in the HER2 gene by cancer type using the published genomic study results, the HER2 mutation frequency was found to be the highest in prostate neuroendocrine cancer, metastatic skin squamous cell carcinoma and bladder cancer, and 2.8 in breast cancer and gastric cancer. % and 9.1% of HER2 mutations were observed (Non-Patent Document 3).

In addition, it has been reported that HER2 signaling activation is improved when a mutation occurs in the kinase domain of the HER2 receptor. Also, it was confirmed that mutations occurring in the same kinase domain have different effects on the reactivity of HER2 targeted therapeutics. For example, the L755S mutation in exon 19 is resistant to trastuzumab and lapatinib but is sensitive to neratinib, and the V777L mutation in exon 20 is resistant to trastuzumab but is resistant to lapatinib and neratinib. It is known to show sensitivity to all nibs, and to show sensitivity to all of trastuzumab, lapatinib, and neratinib in the case of the V842I mutation located in exon 21 (Non-Patent Documents 5 and 6). In addition, for mutations occurring in the extracellular domain affecting HER2 dimerization, G309E increased HER2 dimerization and in the case of S310F, dimerization had no effect, but increased C-terminal tail phosphorylation to induce HER2 signaling activation. Trastuzumab, lapatinib, and neratinib are all known to show sensitivity (Non-Patent Documents 5 and 7). Therefore, there is an urgent need for an effective therapy capable of treating cancer having a HER2 mutation.

Roskoski, R., Jr. (2014) The ErbB/HER family of protein-tyrosine kinases and cancer. Pharmacol Res, 79, 34-74 Zabransky, D.J., Yankaskas, C.L., Cochran, R.L., Wong, H.Y., Croessmann, S., Chu, D., Kavuri, S.M., Red Brewer, M., Rosen, D.M., Dalton, W.B. et al. (2015) HER2 missense mutations have distinct effects on oncogenic signaling and migration. Proc Natl Acad Sci US A, 112, E6205-6214. Connell, C. M. and Doherty, G.J. (2017) Activating HER2 mutations as emerging targets in multiple solid cancers. ESMO Open, 2, e000279. Pahuja, K. B., Nguyen, T. T., Jaiswal, B. S., Prabhash, K., Thaker, T. M., Senger, K., Chaudhuri, S., Kljavin, N. M., Antony, A., Phalke, S. et al. (2018) Actionable Activating Oncogenic ERBB2/HER2 Transmembrane and Juxtamembrane Domain Mutations. Cancer Cell, 34, 792-806 e795. Bose, R., Kavuri, S. M., Searleman, A. C., Shen, W., Shen, D., Koboldt, D. C., Monsey, J., Goel, N., Aronson, A. B., Li, S. et al. (2013) Activating HER2 mutations in HER2 gene amplification negative breast cancer. Cancer Discov, 3, 224-237. Hirotsu, Y., Nakagomi, H., Amemiya, K., Oyama, T., Inoue, M., Mochizuki, H. and Omata, M. (2017) Intrinsic HER2 V777L mutation mediates resistance to trastuzumab in a breast cancer patient . Med Oncol, 34, 3. Greulich, H., Kaplan, B., Mertins, P., Chen, T.H., Tanaka, K.E., Yun, C.H., Zhang, X., Lee, S.H., Cho, J., Ambrogio, L. et al. (2012) Functional analysis of receptor tyrosine kinase mutations in lung cancer identifies oncogenic extracellular domain mutations of ERBB2. Proc Natl Acad Sci U S A, 109, 14476-14481.

One aspect provides an effective amount for treating a cancer comprising at least one mutation present in the kinase domain (KD), mucosal membrane domain (JMD), or transmembrane domain (TMD) of human epidermal growth factor receptor 2 (HER2). To provide a pharmaceutical composition for the treatment of HER2 mutant cancer, including posiotinib.

One aspect provides a pharmaceutical combination for the treatment of a HER2 mutant cancer comprising an effective amount of positiotinib for treating a cancer comprising at least one mutation present in the kinase domain, the mucosal domain, or the transmembrane domain of the HER2 receptor. will provide

Another aspect comprises administering to a subject an effective amount of a pharmaceutical composition comprising positiotinib, which is identified as comprising at least one mutation appearing in a kinase domain, a mucosal membrane domain, or a transmembrane domain of a HER2 receptor. , to provide a method for treating a HER2 mutant cancer.

One aspect comprises at least one mutation appearing in the kinase domain (KD), mucosal membrane domain (JMD), or transmembrane domain (TMD) of human epidermal growth factor receptor 2 (HER2). comprising an effective amount of Poziotinib for the treatment of cancer, wherein the mutation is

exon 21 mutation of HER2;

a mutation selected from V777L, L755S, or a combination thereof in the amino acid sequence of HER2;

a mutation comprising a deletion, insertion, or substitution of 1 to 18 nucleotides at amino acid positions 675 to 714 of HER2; and

a mutation comprising a deletion, insertion, or substitution of 1 to 18 nucleotides at amino acid positions 648 to 674 of HER2; It provides a pharmaceutical composition for the treatment of HER2 mutant cancer, comprising any one selected from combinations thereof.

Another aspect is for treating cancer comprising at least one mutation appearing in the kinase domain (KD), the mucosal membrane domain (JMD), or the transmembrane domain (TMD) of human epidermal growth factor receptor 2 (HER2). an effective amount of positiotinib and an effective amount of a HER2 inhibitor, wherein the mutation comprises

exon 21 mutation of HER2;

a mutation selected from V777L, L755S, or a combination thereof in the amino acid sequence of HER2;

a mutation comprising a deletion, insertion, or substitution of 1 to 18 nucleotides at amino acid positions 675 to 714 of HER2; and

a mutation comprising a deletion, insertion, or substitution of 1 to 18 nucleotides at amino acid positions 648 to 674 of HER2; Including any one selected from these combinations,

It provides a pharmaceutical combination for the treatment of HER2 mutant cancer, wherein the positiotinib and the HER2 inhibitor are administered simultaneously, sequentially or separately.

Another aspect provides a composition comprising positiotinib to a subject suffering from a cancer comprising at least one mutation present in the kinase domain, the mucosal domain, or the transmembrane domain of human epidermal growth factor receptor 2 (HER2). to administer, and the mutation is

exon 21 mutation of HER2;

a mutation selected from V777L, L755S, or a combination thereof in the amino acid sequence of HER2;

a mutation comprising a deletion, insertion, or substitution of 1 to 18 nucleotides at amino acid positions 675 to 714 of HER2; and

a mutation comprising a deletion, insertion, or substitution of 1 to 18 nucleotides at amino acid positions 648 to 674 of HER2; It provides a method of treating HER2 mutant cancer, comprising any one selected from combinations thereof.

The pharmaceutical composition and combination comprising pogiotinib according to an aspect and a method of treating cancer using the same include cancer comprising at least one mutation appearing in the kinase domain, mucosal membrane domain, or transmembrane domain of HER2, such as V777L , L755S, V842I, R678Q, L662Q, or a combination thereof to suppress tumors in cancers, including gastric cancer and breast cancer, having mutations, and inhibit cancer cell survival to show a therapeutic effect of cancer.

In addition, the pharmaceutical composition and the treatment method using the same according to one aspect have limited clinical application of the HER2 targeted therapy because there is no amplification of the HER2 gene despite the presence of HER2 mutation, or have a therapeutic effect on cancer that has developed resistance to the HER2 targeted therapy. indicates.

1 shows changes in tumor volume (mm ³ ) following administration of either posiotinib or trastuzumab alone or in combination in an animal model ( ERBB2 amplified IMX-102 model) established from breast cancer patient tissue. 1A shows a breast cancer model ( ERBB2 amplified IMX-102 model). 1B shows the mapping of the breast cancer model ( ERBB2 amplification IMX-102 model). 1C shows genomic mapping of representative genetic variations identified in a breast cancer model ( ERBB2 amplification IMX-102 model). 1D shows tumor volume (mm ³ ) changes with vehicle, Posiotinib, or Trastuzumab alone or in combination with Posiotinib and Trastuzumab (ER+: estrogen receptor positive breast cancer, ER-: estrogen receptor negative breast cancer). indicates
Figure 2 shows the change in tumor volume (mm ³ ) according to the administration of either posiotinib or trastuzumab alone or in combination in an animal model of gastric cancer having a V842I mutation.
3 shows changes in tumor volume (mm ³ ) following administration of either posiotinib or trastuzumab alone or in combination in an animal model established from a breast cancer patient with L755S or V777L mutation and amplification of the HER2 gene ( ERBB2 ). . 3 The left graph shows changes in the L755S mutant animal model. 3 The graph on the right shows changes in the V777L mutant animal model.
4 shows tumor reduction following administration of Posiotinib in breast cancer patients in which V777L mutation was detected and amplification of the HER2 gene ( ERBB2 ).
Figure 5 shows the change in tumor volume (mm ³ ) according to the administration of posiotinib in an animal model of gastric cancer with R678Q mutation.
6 shows changes in tumor volume (mm ³ ) following administration of either posiotinib or trastuzumab alone or in combination in an animal model of breast cancer with L662Q mutation.
7 shows the results of inducing normal HER2 overexpression and L662Q, R678Q, V777L, or L755S mutations in MCF10A, a normal breast cell line, and verifying the mutations through Sanger sequencing.
Figure 8 shows the cell viability that appears as a result of expressing V777L, L755S, L662Q, or R678Q mutations in the same tumor cell line and administering positiotinib at different concentrations.

Hereinafter, the present invention will be described in more detail.

All technical terms used in the present invention, unless otherwise defined, have the meaning as commonly understood by one of ordinary skill in the art of the present invention. In addition, although preferred methods and samples are described herein, similar or equivalent ones are also included in the scope of the present invention. The contents of all publications incorporated herein by reference are hereby incorporated by reference in their entirety.

1. Definition

As used herein, the singular (“a” or “an”) can mean one or more. As used in the claims, when used in conjunction with the word "comprising", the word "a" or "an" may mean more than one.

The use of the term "or" herein is used to mean "and/or" unless explicitly indicated to refer to alternatives only, or unless alternatives are mutually exclusive, although this disclosure is intended to mean alternatives only and " and/or". As used herein, “another” may mean at least a second or more.

As used herein, the term “treating” or “treatment” refers to inhibiting a disease, e.g., inhibiting a disease, condition or disorder in an individual experiencing or exhibiting the pathology or symptom of a disease, condition or disorder, i.e., inhibiting the disease, condition or disorder; preventing further development of the pathology and/or symptom, or ameliorating the disease, e.g., ameliorating the disease, condition or disorder in an individual experiencing or exhibiting the pathology or symptom of the disease, condition or disorder, i.e., Reversing pathology and/or symptoms, such as reducing disease severity.

As used herein, the term "preventing" or "prevention" refers to preventing a disease, e.g., a disease in an individual who may be predisposed to the disease, condition or disorder but has not yet experienced or exhibited the pathology or signs of the disease; To prevent a condition or disorder.

As used herein, the term "subject", "patient", or "individual" refers to a living mammalian organism, such as a human, monkey, cow, sheep, goat, dog, cat, mouse, rat, guinea pig, or transgenic species thereof. refers to In certain embodiments, the patient or subject is a primate. Non-limiting examples of human patients are adults, adolescents, infants and fetuses.

wherein said subject analyzes a genomic sample from the patient to obtain a mutation appearing in one or more kinase domains selected from the group consisting of an exon 19 mutation, an exon 20 mutation, and an exon 21 mutation; mutations appearing within the mucosal domain; mutations appearing in the transmembrane domain; and individuals identified as having a mutation selected from combinations thereof.

The term “effective” as used herein, as used herein and/or in the claims, means that it is suitable to achieve a desired, expected, or intended result. An “effective amount,” “therapeutically effective amount,” or “pharmaceutically effective amount,” when used in the context of treating a patient or subject with a compound, is when the compound is administered to a subject or patient to treat or prevent the disease, such treatment of a disease. or an amount of the compound that is sufficient to effect prevention.

As used herein, the term "combination" refers to a product produced by mixing or combining one or more active ingredients, and includes both fixed and non-fixed combinations of active ingredients. In the present specification, "fixed combination" is used as known to those skilled in the art, and for example, a first active ingredient and an additional active ingredient (eg, a second active ingredient) are used in one unit dose. It is defined as a combination that exists to be administered as a single entity or as a single entity. An example of the "fixed combination" is a pharmaceutical composition in which a first active ingredient and an additional active ingredient are present as a mixture for simultaneous administration, such as as one formulation. Another example of "fixed combination" is a pharmaceutical combination in which the first active ingredient and the additional active ingredients are not present as a mixture but are present as a unit. For example, a fixed combination refers to a form of administration in which any compound as an active ingredient and a combination partner thereof are both administered to a single entity or patient simultaneously. As used herein, "non-fixed combination" is used as known to a person skilled in the art, and is defined as a combination in which a first active ingredient and an additional active ingredient are present in more than one unit. An example of a non-fixed combination is a combination in which the first active ingredient and the additional active ingredient are separately present. It is possible for the components of the non-fixed combination to be administered individually, sequentially, simultaneously, concurrently or staggered. For example, a non-fixed combination means that any compound as an active ingredient and a combination partner are both administered to a patient simultaneously as separate individuals, in combination, or sequentially without a specific time limit.

For example, the active ingredient or drug included in the combination may be formulated as two, three, or more separate pharmaceutical compositions. For example, one or more active ingredients in the combination are each independently administered as a composition for oral, intravenous, intraarterial, intraperitoneal, intradermal, transdermal, intrathecal, intramuscular, intranasal, transmucosal, subcutaneous or rectal administration. can be formulated. The formulation may be, for example, oral or injection.

As used herein, the term “composition” refers to a pharmaceutical product comprising a therapeutically effective amount of the specified ingredients, as well as any product resulting directly or indirectly from a combination of the specified ingredients in the specified amounts. Composition or pharmaceutical composition also means a mixture comprising at least one compound and at least one pharmaceutically acceptable ingredient such as a carrier, stabilizer, diluent, dispersing agent, suspending agent, thickening agent, or excipient.

As used herein, the term "anticancer" agent reduces tumor size by, for example, promoting the killing of cancer cells, inducing apoptosis in cancer cells, reducing the growth rate of cancer cells, reducing the incidence or number of metastases, By reducing tumor growth, by reducing the blood supply to a tumor or cancer cells, by promoting an immune response against cancer cells or tumors, by preventing or inhibiting the progression of cancer, or by increasing the lifespan of a subject with cancer , negatively affect the cancer cells/tumors of the subject.

As used herein, the term “nucleotide deletion” refers to the removal of a nucleotide at a specific position in a nucleotide sequence. As used herein, "nucleotide insertion" refers to the addition of one or more nucleotide bases to a nucleotide sequence. As used herein, the term “nucleotide substitution” refers to the exchange of a nucleotide by another nucleotide at a specific position in a nucleotide sequence.

As used herein, the term "amino acid deletion" refers to the deletion of one or more amino acid residues in a particular region or region of a polypeptide to reduce the number of amino acid residues in that region of the polypeptide compared to the number of amino acid residues in that region of a native polypeptide. it means. As used herein, "amino acid insertion" means adding at least one amino acid residue to a specific region of a polypeptide to increase the number of amino acid residues in that region compared to the number of amino acid residues in that region of a native polypeptide. As used herein, the term “amino acid substitution” refers to altering the amino acid sequence, not the number of amino acid residues, in a specific region or region of a polypeptide by replacing one or more amino acid residues in the region. Substitution is the result of insertion of another amino acid residue at the same position after deletion of an amino acid residue.

As used herein, the terms “detectable”, “detectable” and grammatical equivalents thereof refer to a method of determining the presence and/or amount and/or identity of a target nucleic acid sequence. In some embodiments, detection occurs by amplification of a target nucleic acid sequence. In other embodiments, sequencing of a target nucleic acid may be characterized by “detection” of the target nucleic acid sequence. The label attached to the probe may comprise any of a variety of different labels known in the art, which may be detected, for example, by chemical or physical means. Labels that may be attached to the probe may include, for example, fluorescent and luminescent materials.

As used herein, the term “pharmaceutically acceptable” refers to tissues, organs, and organs of humans and animals without undue toxicity, irritation, allergic reaction, or other problems or complications proportional to a reasonable benefit/risk ratio, within the scope of sound medical judgment. refers to a compound, substance, composition and/or dosage form suitable for use in contact with bodily fluids.

In this specification, the numerical range indicated using the term “to” includes the range indicated by the lower limit and the upper limit of the numerical values described before and after the term “to”, respectively.

In addition, the numerical values described in this specification are considered to include the meaning of "about" even if not specified. The term "about" is used herein to indicate that a value includes the method utilized to determine the value, the inherent variation in the error of the device, or this variation present in the study subject. The expression "about", or approximately", etc., means that the referenced value may vary to some extent. For example, the value may be 10%, 5%, 2%, or 1 For example, "about 5" is meant to include any value between 4.5 and 5.5, between 4.75 and 5.25, or between 4.9 and 5.1, or between 4.95 and 5.05. .

The terms, “have”, “may have”, “comprises”, or “may include” indicate the presence of the corresponding characteristic (eg, a numerical value or a component such as a component), and of the additional characteristic. Existence is not excluded.

As used herein, “EGFR” or “epidermal growth factor receptor” or “EGFR” refers to a tyrosine kinase cell surface receptor. As used herein, "EGFR" refers to an epidermal growth factor receptor, a type of erbB receptor class protein tyrosine kinase, and a growth factor ligand, such as epidermal growth factor (EGF) binding. can do. EGFR may form a homodimer with another EGFR molecule upon binding to EGF, or may form a heterodimer with another class member, such as erbB2 (HER2), erbB3 (HER3), or erbB4 (HER4). there is. The formation of homo and/or heterodimers of the erbB receptor results in phosphorylation of important tyrosine residues in the intracellular domain and leads to stimulation of many intracellular signaling pathways involved in cell proliferation and survival. Deregulation of erbB class signaling promotes proliferation, invasion, metastasis, angiogenesis and tumor cell survival. For example, EGFR is one of the four alternative transcripts represented by GenBank accession numbers NM _{_} 005228.5, NM _{_} 201282.2, NM _{_} 201283.1, NM _{_} 201284.2, NM_001346897.2, NM_001346898.2, NM_001346899.1 and NM_001346900.2. It can be encrypted by one.

As used herein, "HER2" or "ERBB2" is a member of the EGFR/ErbB family and is represented by GenBank accession numbers NM_004448.3, NM_001005862.2, NM_001289936.1, NM_001289937.1 and NM_001289938.1. As used herein, "HER2" or "ERBB2" refers to human epidermal growth factor receptor 2, and belongs to the epidermal growth receptor group of receptor tyrosine kinase (RTK) as a member of the EGFR/ErbB family. The HER2 receptor consists of an extracellular domain (ECD), a transmembrane domain (TMD), a mucosal membrane domain (JMD), a kinase domain (KD), and a C-terminal tail domain (CTD).

A variant of HER2 comprises a deletion, insertion, substitution, or combination thereof in exon 17, exon 19, exon 20, exon 21, or a combination thereof. HER2 mutations include mutations in the extracellular domain, transmembrane domain, mucosal domain, kinase domain, C-terminal tail domain, and the like. For example, mutations appearing in the kinase domain are known to enhance HER2 signaling activation ( Cancer Discov , 3 , 224-237).

In addition, HER2 mutation is a gene mutation by DNA substitution, addition, deletion, chromosome mutation due to deletion, duplication, or rearrangement of a chromosome fragment, transposon mutation caused by a transposon gene, or a transposon mutation thereof Combinations include deletions, insertions, substitutions, or combinations thereof. For example, the HER2 mutation is one, each independently selected from V777L, L755S, V842I, R678Q, L662Q, A775-G776(ins YVMA), G776VC, G776LC, ΔV777-G778(ins CG), Y835F, and V839G. , two or more deletions, insertions and/or substitutions.

As used herein, the term “HER inhibitor” refers to a substance that inhibits HER activation or function. Examples of HER inhibitors include HER antibodies (eg EGFR, HER2, HER3 or HER4 antibodies); EGFR-targeting drugs; small molecule HER antagonists; HER tyrosine kinase inhibitors; HER2 and EGFR dual tyrosine kinase inhibitors such as lapatinib/GW572016; antisense molecules; and/or substances that bind to or inhibit the function of downstream signaling molecules, such as MAPK or Akt.

As used herein, the term “HER2 inhibitor” refers to a substance that inhibits HER2 activation or function. For example, a HER2 inhibitor is an antibody or small molecule that binds to the HER receptor. For example, HER2 inhibitors or HER2 targeted therapies include monoclonal antibodies such as Trastuzumab and Pertuzumab, and the antibody-drug conjugate Trastuzumab emtansine (T-DM1), and tyrosine kinase TKIs that inhibit ATP binding to the domain include afatinib, lapatinib, neratinib, and the like.

As used herein, the term "patient-derived xenograft technology (PDX)" refers to removing cancer tissue from a patient and transplanting it into an immune-deficient mouse to grow the patient's tumor and then administer various anticancer drugs. technology to check

As used herein, the term "partial response (PR)" refers to a state in which the sum of the long diameters of solid cancers is reduced by 30% or more compared to the underlying lesions after treatment in response to treatment of solid cancer.

2. Posiotinib

Poziotinib is known as a quinazoline-based tyrosine kinase inhibitor that inhibits epidermal growth factor receptors EGFR, HER2, HER3, and HER4. In the composition according to one embodiment, Poziotinib is a small molecule compound that selectively and irreversibly inhibits the EGFR family, including Her1, Her2 and Her4, and inhibits activation and resistance mutants of EGFR and Her2 effect can be shown.

In addition, posiotinib may show a strong cell proliferation inhibitory effect even in cancers with mutations in EGFR and HER2 that induce resistance to TKIs such as gefitinib or afatinib.

3. Pharmaceutical composition

The present specification provides a pharmaceutical composition for treating HER2 mutant cancer, comprising an effective amount of positiotinib for treating cancer comprising at least one mutation appearing in the kinase domain, the mucosal membrane domain, or the transmembrane domain of the HER2 receptor. .

As used herein, the term “mutation” refers to a deletion, addition, or substitution of an amino acid residue in the amino acid sequence of a protein or polypeptide compared to the amino acid sequence of a reference protein or polypeptide. Mutations occurring in living organisms can be divided into gene mutations and chromosomal mutations that occur as chromosomal abnormalities. Gene mutation refers to a mutation caused by structural changes in a gene, which is an intrinsic trait of an individual.

In the present specification, the HER2 mutation may be a gene mutation. The HER2 mutation may include deletion, insertion, or substitution of 1 to 18 nucleotides at a specific position of the amino acid of the HER2 receptor. Alternatively, the mutation may include deletion, insertion, or substitution of 3 to 18 nucleotides at a specific position of the amino acid of the HER2 receptor.

In the present specification, the mutation appearing in the kinase domain of HER2 may have one or more mutations selected from the group consisting of exon 19 mutation, exon 20 mutation, and exon 21 mutation. The mutation appearing in the kinase domain may include, for example, 1 to 6 mutations selected from the group consisting of exon 19 mutation, exon 20 mutation, and exon 21 mutation. The mutation appearing in the kinase domain may include deletion, insertion, or substitution of a nucleotide in at least one selected from the group consisting of exon 19 mutation, exon 20 mutation, and exon 21 mutation.

The mutation appearing in the kinase domain may be an exon 19 mutation such as a mutation at amino acid positions 755 to 769 of the HER2 receptor, an exon 20 mutation mutation such as a mutation at amino acid positions 776 to 816 of the HER2 receptor, or an exon 21 mutation such as HER2 It may be a mutation at amino acid positions 814 to 880 of the receptor, or a combination thereof.

For example, the exon 19 mutation may be any one selected from the group consisting of L755S, L755_T759del, S760A, D769H, and D769Y. For example, the exon 20 mutation may be any one selected from the group consisting of V777L, V777M, P780_Y781insGSP, and Q799M. For example, the exon 21 mutation may be any one selected from the group consisting of V842I, L834R, V842I, and L869Q.

Mutations appearing in the kinase domain of HER2 herein may include substitution, insertion, or deletion of 1 to 18 nucleotides between amino acids 755 to 869. or substitutions, insertions, or deletions of 3 to 18 nucleotides at the position. The mutation appearing in the kinase domain may include two or more mutations selected from the group consisting of V777L, L755S, and V842I.

Mutations appearing in the mucosal domain of HER2 herein may include substitution, insertion, or deletion of 1 to 18, or 3 to 18 nucleotides between amino acids 675 to 714. Alternatively, the mutation may include substitution, insertion, or deletion of 3 to 18 nucleotides between amino acids 677 to 683.

Mutations appearing in the transmembrane domain of HER2 herein may include substitution, insertion, or deletion of 1 to 18, or 3 to 18 nucleotides between amino acids 648 to 674.

In the present specification, the HER2 mutation may include deletion, insertion, or substitution of one or more nucleotides at amino acid positions 662, 678, 755, 760, 769, 777, 780, 799, 834, 842, or 869. The HER2 mutation may be V777L, L755S, V842I, R678Q, L662Q, or a combination thereof.

One aspect comprises at least one mutation appearing in the kinase domain (KD), mucosal membrane domain (JMD), or transmembrane domain (TMD) of human epidermal growth factor receptor 2 (HER2). comprising an effective amount of Poziotinib for the treatment of cancer, wherein the mutation is

exon 21 mutation of HER2;

a mutation selected from V777L, L755S, or a combination thereof in the amino acid sequence of HER2;

a mutation comprising a deletion, insertion, or substitution of 1 to 18 nucleotides at amino acid positions 675 to 714 of HER2; and

a mutation comprising a deletion, insertion, or substitution of 1 to 18 nucleotides at amino acid positions 648 to 674 of HER2; It provides a pharmaceutical composition for the treatment of HER2 mutant cancer, comprising any one selected from combinations thereof.

The pharmaceutical composition comprising positiotinib may exhibit therapeutic effects on cancer in which the HER2 signaling system is activated by HER2 mutation as well as HER2 gene amplification. The HER2 mutation may include at least one mutation appearing in a kinase domain, a mucosal membrane domain, or a transmembrane domain of a HER2 receptor.

In one embodiment, the exon 21 mutation of HER2 may include a deletion, insertion, or substitution of 1 to 18 nucleotides at positions 824 to 875 of amino acids of HER2. For example, the exon 21 mutation may be, for example, a mutation at amino acid positions 824 to 875, a mutation at positions 830 to 845, or a mutation at positions 830 to 869 of the HER2 receptor. In addition, the exon 21 mutation of HER2 may include a deletion, insertion, or substitution of 1 to 18 or 3 to 18 nucleotides at the amino acid position.

In one embodiment, the mutation of HER2 may include one or more deletions, insertions, or substitutions at amino acid positions 662, 678, 755, 777, or 842.

In one embodiment, the mutation appearing in the mucosal domain may include substitution, insertion, or deletion of 1 to 18 nucleotides at amino acid positions 677 to 683.

In one embodiment, the mutation appearing in the transmembrane domain may comprise a substitution, insertion, or deletion of 1 to 18 nucleotides at amino acid positions 653 to 662.

In one embodiment, the mutation appearing in the mucosal domain may comprise a substitution, insertion, or deletion of a nucleotide at amino acid 678 of HER2.

In one embodiment, the mutation appearing in the transmembrane domain may comprise a substitution, insertion, or deletion of a nucleotide at amino acid 662 of HER2.

In one embodiment, the mutation appearing in the kinase domain may comprise two or more mutations selected from the group consisting of V777L, L755S, and V842I.

In one embodiment, the mutation appearing in the mucosal domain may be R678Q.

In one embodiment, the mutation appearing in the transmembrane domain may be L662Q.

In one embodiment, the exon 21 mutation may be any one selected from the group consisting of L834R, V842I, and L869Q.

In one embodiment, it may further comprise an effective amount of a HER2 inhibitor for treating the cancer.

In one embodiment, the positiotinib may be administered in combination with the HER2 inhibitor simultaneously, sequentially or separately.

In one embodiment, the HER2 inhibitor is Trastuzumab, Pertuzumab, Trastuzumab emtansine (T-DM1), Afatinib (BIBW2992), Lapatinib, yes Latinib (Neratinib), trastuzumab deruxtecan (Trastuzumab deruxtecan) (DS-8201a), or may be selected from a combination thereof.

The pharmaceutical composition provides a preventive or therapeutic use for cancer. In one embodiment, it may be for the treatment of gastric cancer or breast cancer.

The pharmaceutical composition according to one embodiment may be a pharmaceutical composition for treating gastric cancer having any one mutation selected from V842I, R678Q, or a combination thereof.

The pharmaceutical composition according to one embodiment may be a pharmaceutical composition for treating breast cancer having any one mutation selected from V777L, L755S, L662Q, or a combination thereof.

The dosage of the pharmaceutical composition is an amount effective for treating or inhibiting recurrence of metastatic cancer in an individual or patient, and may be administered orally or parenterally according to the purpose. When administered parenterally so as to be administered in an amount of 0.01 to 1000 mg/kg, more specifically 0.1 to 300 mg/kg, 0.01 to 100 mg/kg of body weight per day, more specifically 0.1 to 50 mg/kg, based on the active ingredient It can be administered in 1 to several divided doses to be administered in an amount of The dose to be administered to a specific individual or patient should be determined in light of several related factors such as the patient's weight, age, sex, health condition, diet, administration time, administration method, and disease severity, and can be appropriately adjusted or decreased by a specialist. It should be understood that the above dosage is not intended to limit the scope of the present invention in any way. A physician or veterinarian of ordinary skill in the art can readily determine and prescribe the required effective amount of the pharmaceutical composition. For example, a physician or veterinarian may start a dose of a compound of the invention used in a pharmaceutical composition at a level lower than that required to achieve the desired therapeutic effect, and gradually increase the dosage until the desired effect is achieved. can increase

The pharmaceutical composition may include conventional pharmaceutically acceptable carriers, excipients or additives. The pharmaceutical composition may be formulated according to a conventional method, and various oral dosage forms such as tablets, pills, powders, capsules, syrups, emulsions, microemulsions, etc. or parenteral administration such as intramuscular, intravenous, intraperitoneal or subcutaneous administration may be prepared in oral dosage forms. The parenteral dosage form may be an injection.

The pharmaceutical composition may further include one or more optional pharmaceutically acceptable additives selected from the group consisting of excipients, binders, disintegrants, lubricants, and any combination thereof. The excipient is any substance known to those skilled in the art to be useful for preparing formulations, and may be adjusted as necessary, for example, according to the mode of administration of the drug.

When the pharmaceutical composition is prepared in the form of an oral dosage form, examples of additives or carriers used include cellulose, calcium silicate, corn starch, lactose, sucrose, dextrose, calcium phosphate, stearic acid, magnesium stearate, stearic acid calcium, gelatin, talc, surfactant, suspending agent, emulsifying agent, diluent, and the like. When the pharmaceutical composition is prepared in the form of an injection, the additive or carrier includes water, saline, aqueous glucose solution, similar sugar solution, alcohol, glycol, ether (eg, polyethylene glycol 400), oil, fatty acid, fatty acid ester, glycer. lides, surfactants, suspending agents, emulsifiers, and the like.

The pharmaceutical composition may be administered alone or in combination with one or more additional therapeutic agents. "Combination administration" refers to administration of a pharmaceutical composition according to one aspect and one or more additional therapeutic agents to the subject or patient to be treated, either simultaneously or at different time points and in any order. Accordingly, each component may be administered separately, or may be administered in sufficiently close proximity to provide the desired therapeutic effect. The one or more additional therapeutic agents may be, for example, other anti-cancer agents.

4. Pharmaceutical Combinations

Another aspect provides an effective amount for treating a cancer comprising at least one mutation appearing in the kinase domain (KD), mucosal membrane domain (JMD), or transmembrane domain (TMD) of human epidermal growth factor receptor 2 (HER2). Posiotinib and an effective amount of a HER2 inhibitor, wherein the mutation is

exon 21 mutation of HER2;

a mutation selected from V777L, L755S, or a combination thereof in the amino acid sequence of HER2;

a mutation comprising a deletion, insertion, or substitution of 1 to 18 nucleotides at amino acid positions 675 to 714 of HER2; and

a mutation comprising a deletion, insertion, or substitution of 1 to 18 nucleotides at amino acid positions 648 to 674 of HER2; Including any one selected from these combinations,

It provides a pharmaceutical combination for the treatment of HER2 mutant cancer, wherein the positiotinib and the HER2 inhibitor are administered simultaneously, sequentially or separately.

In one embodiment, the HER2 inhibitor may be selected from trastuzumab, pertuzumab, trastuzumab emtacin, afatinib, lapatinib, neratinib, trastuzumab deluxtecan, or a combination thereof.

In one embodiment, the positiotinib may be an oral dosage form, and the HER2 inhibitor may be an oral dosage form or an injection dosage form. The injection formulation may be, for example, intravenous (i.v), intraperitoneal (i.p), or subcutaneous (s.c) injection, and may be administered alternately if necessary. In addition, the injection formulation may be administered by instillation or bolus, if necessary. For example, the injectable formulation may be instilled over about 20 to 120 minutes, such as about 30 to 90 minutes, or infused at a time in 5 to 10 minutes.

In the pharmaceutical composition, pogiotinib may be included in a dose of 0.5 to 100 mg/day. Pogiotinib in the pharmaceutical composition may be, for example, a dose of 1 to 30 mg/day, a dose of 3 to 25 mg/day, a dose of 3 to 20 mg/day, or a dose of 6 to 16 mg/day. . In addition, in the pharmaceutical composition, the HER2 inhibitor may be included in a dose of 0.1 to 300 mg/kg/week based on the body weight (kg) of the administered subject.

In one embodiment, posiotinib is included in a dose of 0.05 to 10 mg/kg/day based on the body weight (kg) of the administered subject, and the HER2 inhibitor is 1 to 250 mg based on the body weight (kg) of the administered subject It can be included in a dose of /kg/week.

Alternatively, in the pharmaceutical composition, pogiotinib may be administered in a dose of 0.05 to 5 mg/kg/day, for example, 0.1 to 1 mg/kg/day, 0.1 to 0.5 mg/day, based on the body weight (kg) of the administered subject. kg/day dose, or 0.5 to 5 mg/kg/day dose, wherein the HER2 inhibitor is administered at a dose of 0.1 to 250 mg/kg/week, for example 1 to 250 mg/kg/week, based on the body weight (kg) of the administered subject It may be included in a dose of 200 mg/kg/week.

In another embodiment, positiotinib is included in a dose of 1 to 100 mg/kg/3 weeks based on the body weight (kg) of the administered subject, and the HER2 inhibitor is 3 to 750 based on the body weight (kg) of the administered subject It may be included in a dose of mg/kg/3 weeks. For example, posiotinib may be included in a dose of 1 to 20 mg/kg/3 weeks, trastuzumab as a HER2 inhibitor may be in a dose of 3 to 20 mg/kg/3 weeks, and afatinib as a HER2 inhibitor is 0.5 to 10 mg/kg/day dose, lapatinib as HER2 inhibitor may be 1-30 mg/kg/day dose, and neratinib as HER2 inhibitor 1-10 mg/kg/day dose can be

Or, for example, posiotinib may be administered at a dose of 0.05 to 10 mg/kg/day, at a dose of 0.05 to 5 mg/kg/day, or between 0.5 and 5 mg/kg/1, based on the body weight (kg) of the administered subject. It may be included in a daily dose. For example, the daily dosage of posiotinib may be 5 to 24 mg. For example, upon continuous administration, the maximum tolerated dose (MTD) of pogiotinib may be 25 mg or less, such as 18 mg. Also, when administered intermittently, the maximum tolerated dose (MTD) of pogiotinib may be 24 mg.

Alternatively, for example, 1 to 25 mg of Posiotinib may be administered qd (daily) during the dosing period. For example, the daily dose of pogiotinib may be 4 mg, 8 mg, 10 mg, 12 mg, 16 mg, 18 mg, or 24 mg. For example, by administering posiotinib (12 mg, qd, p.o) for 14 days and resting for 7 days, each cycle can be configured for a total of 21 days (3 weeks). In addition, it is possible to establish a dosing plan by observing whether the tumor is reduced after drug administration. The medication regimen is until disease progression or intolerable toxicity. Progression-free survival (PFS) can be evaluated as a primary endpoint. In addition, objective response rate (ORR), overall survival (OS), and safety can be evaluated.

Or, for example, the HER2 inhibitor may be administered at a dose of 0.1 to 250 mg/kg/week, a dose of 0.1 to 200 mg/kg/week, or a dose of 1-200 mg/kg/week, based on the body weight (kg) of the administered subject. , 1 to 150 mg/kg/week dose, 1 to 100 mg/kg/week dose, 1 to 50 mg/kg/week dose, or 2 to 20 mg/kg/week dose, 0.1 to 50 mg It may be included in a dose of /kg/week, a dose of 1 to 25 mg/kg/week, or a dose of 2 to 20 mg/kg/week. The HER2 inhibitor may be trastuzumab. For example, the HER2 inhibitor may be administered on a one-week regimen or three-week regimen. For example, the recommended initial loading dose of trastuzumab for 1 week is 4 mg/kg, and the recommended maintenance dose is 2 mg/kg weekly, and administration is started one week after the administration of the initial loading dose. For example, the recommended initial loading dose of trastuzumab for 3 weeks is 8 mg/kg, and thereafter, the recommended maintenance dose is 6 mg/kg every 3 weeks thereafter, and administration starts 3 weeks after the administration of the initial loading dose. For example, 10 to 2000 mg of the HER2 inhibitor may be administered qw (once a week) during the dosing period.

As for the pharmaceutical combination and the pharmaceutical composition, the description or exemplary description for any one may be applied as it is unless there is a description to the contrary.

5. Treatment method, use

Another aspect is

administering a composition comprising positiotinib to a subject suffering from a cancer comprising at least one mutation appearing in the kinase domain, the mucosal membrane domain, or the transmembrane domain of human epidermal growth factor receptor 2 (HER2), The mutation is

exon 21 mutation of HER2;

a mutation selected from V777L, L755S, or a combination thereof in the amino acid sequence of HER2;

a mutation comprising a deletion, insertion, or substitution of 1 to 18 nucleotides at amino acid positions 675 to 714 of HER2; and

a mutation comprising a deletion, insertion, or substitution of 1 to 18 nucleotides at amino acid positions 648 to 674 of HER2; It provides a method of treating HER2 mutant cancer, comprising any one selected from combinations thereof.

The cancer may be a cancer having an EGFR mutation. The EGFR mutation may be an activating EGFR mutation, a mutation inducing resistance to an EGFR inhibitor, or a combination thereof. The cancer may be a cancer having a HER2 mutation.

For example, the cancer is lung cancer, liver cancer, esophageal cancer, stomach cancer, colon cancer, small intestine cancer, pancreatic cancer, melanoma, breast cancer, oral cancer, brain tumor, thyroid cancer, parathyroid cancer, kidney cancer, cervical cancer, sarcoma, prostate cancer, urethral cancer , bladder cancer, testicular cancer, blood cancer, lymphoma, skin cancer, may be selected from the group consisting of psoriasis and fibroadenoma.

The method may have therapeutic effects on cancers that do not show amplification of the HER2 gene. The method exhibits therapeutic effects on cancers that have limited clinical application of HER2 targeted therapies or have developed resistance to HER2 targeted therapies due to lack of amplification of the HER2 gene despite the presence of HER2 mutations. For example, the method provides for use in treating gastric or breast cancer. For example, the method provides for the treatment of gastric or breast cancer having at least one HER2 mutation selected from the group consisting of V777L, L755S, V842I, R678Q, and L662Q.

The method may be used for the treatment of gastric cancer having any one mutation selected from V842I, R678Q, or a combination thereof.

The method may be used for the treatment of breast cancer having any one mutation selected from V777L, L755S, L662Q, or a combination thereof.

The above method can be utilized as an effective next-generation therapy when resistance to HER2-targeted therapies, trastuzumab or lapatinib, develops, and clinical application of positiotinib in cancers with HER2 exon 19, 20, and 21 mutations offer possibilities

In the method, the description of the pharmaceutical composition may be applied as it is. In addition, the dosage used in the prophylaxis or treatment method is an amount effective for treatment or prevention of an individual or patient, and the description of the dosage of the pharmaceutical composition or pharmaceutical combination may be applied as it is.

Routes of administration of the method include, but are not limited to, oral, intravenous, intraarterial, intraperitoneal, intradermal, transdermal, intrathecal, intramuscular, intranasal, transmucosal, subcutaneous and rectal administration.

The daily dose of the drug according to the above method may be administered as a single dose or divided dose, or may be given as a continuous infusion in the case of parenteral administration. Alternatively, the administration interval may be several seconds, several minutes, several hours, or the number of days at a predetermined interval, and may have a pause if necessary.

For example, the method of treatment comprises positiotinib or a HER2 inhibitor in combination with at least one additional therapy. Additional therapies include radiation therapy, surgery (eg, mastectomy and mastectomy), chemotherapy, gene therapy, DNA therapy, viral therapy, RNA therapy, immunotherapy, bone marrow transplantation, nanotherapy, monoclonal antibody therapy, or any of the foregoing. may be a combination of The additional therapy may be in the form of adjuvant or renal adjuvant therapy.

For example, the additional therapy may be the administration of a small molecule inhibitor or an antimetastatic agent. Alternatively, it may be administration of a side-effect limiting agent (eg, an agent intended to reduce the occurrence and/or severity of side effects of treatment, eg, anti-nausea agents, etc.). Alternatively, it may be radiation therapy, gamma irradiation, surgery, or a combination of radiation therapy and surgery. Alternatively, the additional therapy may be one or more chemotherapeutic agents known in the art.

Posiotinib or a HER2 inhibitor, such as trastuzumab, may be administered before, during, after, or in various combinations for additional cancer therapy, such as immune checkpoint therapy. Administration can be simultaneous to at intervals ranging from minutes, days, weeks. In embodiments in which positiotinib or a HER2 inhibitor is provided to the patient separately from the additional therapeutic agent, the time period between each delivery may be adjusted so that both compounds can still exert a beneficially combined effect on the patient. In this case, antibody therapy and anticancer therapy may be provided to the patient within about 0.5 to 72 hours of each other, or within about 12 to 24 or 72 hours of each other, specifically, within about 0.5 to 12 hours of each other. In some cases, several days (2, 3, 4, 5, 6, or 7) to several weeks (1, 2, 3, 4, 5, 6, 7, or 8 weeks) elapse between each administration. It can be adjusted, and if necessary, there may be a rest period of 1 to 21 days between drug administration and drug administration. For example, posiotinib may have a one-week break after two weeks of administration, and if necessary, one day off after one day of dosing, one day off after two days of dosing, one day off after three days of dosing, one day off after three days of dosing, and four days off after administration. The dosing schedule can be adjusted by taking a 1-day break after taking the drug, and a 2-day break after taking the drug for 5 days. In addition, the HER2 inhibitor, such as trastuzumab, may be administered once every 3 weeks, and the dosing schedule may be adjusted, for example, with a 3-week break after administration for 1 day.

Various embodiments may be used in combination therapy with posiotinib or a HER2 inhibitor, such as trastuzumab. In the examples below, positiotinib is "A" and a HER2 inhibitor, such as trastuzumab, is "B":

A/B/A B/A/B B/B/A A/A/B A/B/B B/A/A A/B/B/B B/A/B/B

B/B/B/A B/B/A/B A/A/B/B A/B/A/B A/B/B/A B/B/A/A

B/A/B/A B/A/A/B A/A/A/B B/A/A/A A/B/A/A A/A/B/A

Administration of any compound or therapy to a patient in the above methods of treatment may follow the general protocol for administration of such compounds, taking into account the toxicity of such agents (if any). For example, it may additionally include monitoring for toxicity attributed to the combination therapy.

In one embodiment, the method of treating cancer may be to additionally administer an effective amount of a HER2 inhibitor for treating the cancer.

In one embodiment, positiotinib and the HER2 inhibitor may be administered simultaneously, sequentially or separately.

In the present specification, the drug is administered 5 times a day, 4 times a day, tid (3 times a day), bid (2 times a day), qd (daily), qod (every 2 days), biw (weekly) according to the administration plan twice a week), tiw (three times a week), qw (once a week), qow (every two weeks), three times a month or once a month.

In one embodiment, the HER2 inhibitor is selected from trastuzumab, pertuzumab, trastuzumab emtacin, afatinib, lapatinib, neratinib, trastuzumab deluxtecan, (DS-8201a), or a combination thereof. it may be

For example, the positiotinib is administered at a dose of 0.05 to 5 mg/kg/day based on the body weight (kg) of the administered subject, and a HER2 inhibitor, such as trastuzumab, is administered based on the body weight (kg) of the subject. It may be administered at a dose of 1 to 50 mg/kg/week.

In one embodiment, the subject analyzes a genomic sample from the patient

exon 21 mutation of HER2;

a mutation selected from V777L, L755S, or a combination thereof in the amino acid sequence of HER2;

a mutation comprising a deletion, insertion, or substitution of 1 to 18 nucleotides at amino acid positions 675 to 714 of HER2; and

a mutation comprising a deletion, insertion, or substitution of 1 to 18 nucleotides at amino acid positions 648 to 674 of HER2; It may be a subject identified as including any one selected from combinations thereof.

In one embodiment, the genomic sample may be isolated from fluid, blood, urine, normal tissue or tumor tissue.

In one embodiment, the presence of the mutation may be determined by nucleic acid sequencing or PCR analysis.

Another aspect is

comprising co-administering an effective amount of posiotinib and an effective amount of a HER2 inhibitor to the subject;

A method for treating cancer comprising at least one mutation present in the kinase domain (KD), mucosal membrane domain (JMD), or transmembrane domain (TMD) of a HER2 receptor, the method comprising:

The mutation is

exon 21 mutation of the HER2 receptor;

a mutation comprising a deletion, insertion, or substitution of 3 to 18 nucleotides at amino acid positions 662 to 678 of the HER2 receptor;

a mutation in the kinase domain of the HER2 receptor selected from V777L, L755S, or a combination thereof; and any one selected from combinations thereof,

The positiotinib and the HER2 inhibitor may be administered simultaneously, sequentially or separately, for a method for treating a HER2 mutant cancer.

In one embodiment, the HER2 inhibitor may be selected from trastuzumab, pertuzumab, trastuzumab  emtacin, afatinib, lapatinib, neratinib, or a combination thereof.

6. Combination Administration Composition

Another aspect is a pharmaceutical composition for treating cancer comprising poziotinib,

An effective amount of Poziotinib for treating a cancer comprising at least one mutation present in the kinase domain (KD), mucosal membrane domain (JMD), or transmembrane domain (TMD) of a HER2 receptor, said mutation is

exon 21 mutation of the HER2 receptor;

a mutation comprising a deletion, insertion, or substitution of 3 to 18 nucleotides at amino acid positions 662 to 678 of the HER2 receptor; and

a mutation in the kinase domain of the HER2 receptor selected from V777L, L755S, or a combination thereof; It will include any one selected from

Provided is a pharmaceutical composition wherein posiotinib is administered in combination with a HER2 inhibitor.

In one embodiment, the HER2 inhibitor administered in combination with pogiotinib may be selected from trastuzumab, pertuzumab, trastuzumab emtacin, afatinib, lapatinib, neratinib, DS-8201a, or a combination thereof. there is.

Hereinafter, the present invention will be described in detail based on the following examples. However, the following examples are only for illustrating the present invention, and the scope of the present invention is not limited thereto.

[Example]

Preparation Example 1. Establishment of a cancer patient tissue-derived animal model (PDX)

A patient-derived cancer tissue xenograft (PDX) animal model was established (BX46, V842I; XT135, V777L) from the tissues of a gastric cancer patient with a V842I mutation in the HER2 kinase domain (KD) and a breast cancer patient with a V777L mutation (BX46, V842I; XT135, V777L). Cell lines (SNU-4256, V842I) and organoids (SNU-5047-TO, V842I, SNU-4842-TO, V777L) were established. In addition, a PDX model was established from the tissue of a breast cancer patient with an L755S mutation in the kinase domain (BX145).

A PDX animal model (BX53) was established from the tissue of a gastric cancer patient with an R678Q mutation in the mucosal membrane domain (JMD), and a cell line (SNU-4257) was established from the cancer tissue of the animal model.

A PDX animal model (BMX214) was established from the tissue of a breast cancer patient having an L662Q mutation in the transmembrane domain (TMD), and a cell line (SNU-4404) was established from the tissue of the animal model.

In addition, two gastric cancer patients with HER2 kinase domain and mucosal domain mutations, breast cancer patients with transmembrane domain mutations without HER2 gene amplification, and HER2 gene amplification and mutations in the kinase domain occurred after treatment with trastuzumab and lapatinib. A total of 5 PDX models were established by cutting the tissues obtained from 2 patients with resistance into 2 mm and transplanting them into the flank adipose tissue of 6-week-old Severe Combined Immunodeficiency Female (NOG) mice.

Experimental Example 1. Evaluation of drug reactivity in cancer patient tissue-derived animal model (PDX)

In the PDX model established from cancer patient tissue, when the tumor grows to 1.5 cm, Posiotinib (1 mg/kg, qd, p.o) and Trastuzumab (10 mg/kg, qw, i.p) are administered alone, divided into 4 groups of 5 animals each. Alternatively, co-administration was used to evaluate drug reactivity. The drug was administered for a total of 28 days, posiotinib was orally administered once daily, and trastuzumab was administered intraperitoneally once a week. Posiotinib was provided by Hanmi Pharmaceutical Co., Ltd. and used, and for Trastuzumab, commercially available Herceptin® was used.

Tumor size was measured every two days to evaluate the degree of tumor growth inhibition, and the stability of the drug was evaluated through animal weight change and behavioral observation.

1 shows changes in tumor volume (mm ³ ) following administration of either posiotinib or trastuzumab alone or in combination in an animal model ( ERBB2 amplified IMX-102 model) established from breast cancer patient tissue. As shown in FIG. 1 , it was confirmed that posiotinib inhibited tumor growth in a patient-derived animal model (PDX) established from skin metastases of HER2 gene amplified breast cancer patients.

Experimental Example 2. Confirmation of HER2 mutation in cancer patient tissue-derived animal model (PDX)

In the PDX model established from gastric cancer or breast cancer patients receiving treatment at Seoul National University Hospital, it was confirmed that there were mutations in two of the gastric cancer models and three of the breast cancer models through whole exon genome analysis (Table 1).

[Table 1]

Table 1 shows the HER2 mutations (V777L, L755S, V842I, R678Q, and L662Q) identified in gastric or breast cancer models. Among them, a total of three mutations in the kinase domain were found, and V842I, V777L, and L755S were found in gastric cancer models (V842I) or breast cancer models (V777L, L755S). In addition, the R678Q mutation in the mucosal domain was found in a gastric cancer model. In addition, a new mutation in the transmembrane domain, the L662Q mutation, was found in a breast cancer model. In particular, in the case of two breast cancer models with mutations (V777L, L755S) in the kinase domain, HER2 gene amplification was simultaneously found, and mutations were found in the tissues at the time when resistance was developed after administration of lapatinib or trastuzumab. It is thought to be related to resistance.

Example 1. Evaluation of drug reactivity in an animal model (PDX) derived from gastric cancer patient tissue with HER2 mutation

In a gastric cancer PDX model with V842I, a mutation known to activate the HER2 signaling system in the kinase domain without HER2 gene amplification, the tumor growth inhibitory ability of Posiotinib and Trastuzumab alone or in combination was evaluated.

A V842I mutation was induced in the kinase domain using overlap extension PCR in a normal breast cell line (MCF-10A). Then, the mutation was verified through Sanger Sequencing, and the degree of inhibition of cell proliferation by positiotinib, trastuzumab, or a combination thereof was evaluated, respectively.

Figure 2 shows the change in tumor volume (mm ³ ) according to the administration of either posiotinib or trastuzumab alone or in combination in an animal model of gastric cancer having a V842I mutation. As shown in FIG. 2 , the gastric cancer PDX model with the V842I mutation showed resistance to trastuzumab, but it was found that it was very effective in the administration of posiotinib alone or the combination of posiotinib and trastuzumab.

Example 2. Evaluation of drug responsiveness in an animal model (PDX) derived from breast cancer patient tissue with HER2 mutation

To evaluate the difference in the reactivity of posiotinib according to HER2 mutation, L755S and V777L mutations were induced in the kinase domain using overlap extension polymerase chain reaction (overlap extension PCR) in MCF10A, a normal breast cell line, and R678Q in the mucosal membrane domain mutations and L662Q mutations in the transmembrane domain. Then, the mutation was verified by Sanger sequencing. In each mutant-expressing cell line, the normal HER2 overexpressing cell line, and the normal HER2 expressing cell line, the degree of inhibition of cell proliferation by positiotinib was evaluated, respectively.

3 shows changes in tumor volume (mm ³ ) following administration of either posiotinib or trastuzumab alone or in combination in an animal model established from a breast cancer patient with L755S and V777L mutations and amplification of the HER2 gene ( ERBB2 ). . As shown in FIG. 3 , it was confirmed that Posiotinib alone was more effective than Trastuzumab in a breast cancer model with exon 19 and exon 20 mutations L755S and V777L mutations and HER2 gene amplification. In addition, particularly when there is a V777L mutation, it exhibited strong activity such as inducing tumor regression.

Example 3. Clinical results according to drug administration in breast cancer patients with HER2 mutation

A V777L mutation was detected in exon 20 of HER2 in a 48-year-old breast cancer patient with an amplification of the HER2 gene who developed resistance to Trastuzumab and Trastuzumab emtansine (T-DM1). Posiotinib (12mg, qd, p.o) was administered for 14 days, and tumor reduction was observed while resting for 7 days.

4 shows tumor reduction following administration of Posiotinib in breast cancer patients in which V777L mutation was detected and amplification of the HER2 gene ( ERBB2 ). As shown in Figure 4, it showed a partial response (PR) showing a marked tumor reduction in 3 cycles of Posiotinib administration.

Example 4. Evaluation of tumor suppression in an animal model (PDX) derived from gastric cancer patient tissue with HER2 mutation

Figure 5 shows changes in tumor volume (mm ³ ) according to drug administration of posiotinib (1 mg/kg, qd, po) for 21 days in an animal model of gastric cancer with R678Q mutation. As shown in FIG. 5 , it can be confirmed that even in the case of having the R678Q mutation occurring in the mucosal membrane domain in gastric cancer, the cell proliferation inhibitory ability by posiotinib is shown.

Example 5. Evaluation of tumor suppression in an animal model (PDX) derived from tissue from breast cancer patients with HER2 mutations

Figure 6 shows tumor volume (mm ³ ) according to posiotinib (1mg/kg, qd, po) and trastuzumab (10mg/kg, qw, ip) administered alone or in combination for 28 days in an animal model of breast cancer with L662Q mutation. ) represents a change. As shown in FIG. 6 , it was confirmed that tumor growth was effectively inhibited by positiotinib in cancers having L662Q mutations occurring in the transmembrane domain.

Example 6. Sanger sequencing results of breast cell lines with HER2 mutation

7 is a normal breast cell line, MCF10A, normal HER2 overexpression and L662Q, R678Q, V777L, or L755S mutations are induced through gene introduction using a retroviral vector, and genomic DNA is extracted from the cell line (Qiagen, QIAamp DNa The results verified through Sanger sequencing using mini kit #51306) primers (Forward 5`-CTTTGGGAGCCTGGCATTTC-3`; Reverse 5`-GCATACACCAGTTCAGCAGG-3`) are shown.

Example 7. Confirmation of apoptosis effect according to administration of posiotinib in a cell line having a HER2 mutation

8 is a cell line expressing V777L, L755S, L662Q, or R678Q mutations in the same normal breast cell line MCA10A, seeding 3000 to 8000 cells in a 96 well plate, 24 hours later, varying concentrations from 0.1 to 100 nm, The viability of cells appearing 5 days after administration of the nib is shown.

As shown in FIG. 8, after expressing different HER2 in the same cell line, the degree of inhibition of cell survival by positiotinib was evaluated. As a result, not only V777L and L755S mutations occurring in the kinase domain, but also L662Q mutations in the transmembrane domain and the mucosal domain It was confirmed that all of the R678Q mutations of posiotinib inhibit cell survival.

So far, the present invention has been looked at with respect to preferred embodiments thereof. Those of ordinary skill in the art to which the present invention pertains will understand that the present invention can be implemented in a modified form without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments are to be considered in an illustrative rather than a restrictive sense. The scope of the present invention is indicated in the claims rather than the foregoing description, and all differences within the scope equivalent thereto should be construed as being included in the present invention.

Claims (28)

Treating cancer comprising at least one mutation present in the kinase domain (KD), mucosal membrane domain (JMD), or transmembrane domain (TMD) of human epidermal growth factor receptor 2 (HER2) comprising an effective amount of Poziotinib for
exon 21 mutation of HER2;
a mutation selected from V777L, L755S, or a combination thereof in the amino acid sequence of HER2;
a mutation comprising a deletion, insertion, or substitution of 1 to 18 nucleotides at amino acid positions 675 to 714 of HER2; and
a mutation comprising a deletion, insertion, or substitution of 1 to 18 nucleotides at amino acid positions 648 to 674 of HER2; Which includes any one selected from combinations thereof,
A pharmaceutical composition for the treatment of HER2 mutant cancer. The pharmaceutical composition of claim 1, wherein the exon 21 mutation of HER2 comprises a deletion, insertion, or substitution of 1 to 18 nucleotides at amino acid positions 824 to 875 of HER2. The pharmaceutical composition according to claim 1, wherein the mutation of HER2 comprises one or more deletions, insertions or substitutions at amino acid positions 662, 678, 755, 777, or 842. The composition of claim 1, wherein the mutation appearing in the mucosal domain comprises substitution, insertion, or deletion of 1 to 18 nucleotides at amino acid positions 677 to 683. The composition of claim 1, wherein the mutation appearing in the transmembrane domain comprises substitution, insertion, or deletion of 1 to 18 nucleotides at amino acid positions 653 to 662. The composition of claim 1 , wherein the mutation appearing in the mucosal domain comprises a substitution, insertion, or deletion of a nucleotide at amino acid 678 of HER2. The composition of claim 1 , wherein the mutation appearing in the transmembrane domain comprises a substitution, insertion, or deletion of a nucleotide at amino acid 662 of HER2. The pharmaceutical composition according to claim 1, wherein the mutation appearing in the kinase domain comprises two or more mutations selected from the group consisting of V777L, L755S, and V842I. The pharmaceutical composition according to claim 1, wherein the mutation appearing in the mucosal membrane domain is R678Q. The pharmaceutical composition according to claim 1, wherein the mutation appearing in the transmembrane domain is L662Q. The pharmaceutical composition of claim 1, wherein the exon 21 mutation is any one selected from the group consisting of L834R, V842I, and L869Q. The pharmaceutical composition of claim 1, further comprising an effective amount of a HER2 inhibitor for treating the cancer. The pharmaceutical composition according to claim 1, wherein the posiotinib is administered in combination with the HER2 inhibitor simultaneously, sequentially or separately. The method according to claim 12 or 13, wherein the HER2 inhibitor is Trastuzumab, Pertuzumab, Trastuzumab emtansine (T-DM1), Afatinib (BIBW2992), Lapatinib , neratinib (Neratinib), trastuzumab deruxtecan (Trastuzumab deruxtecan) (DS-8201a), or a pharmaceutical composition selected from a combination thereof. The pharmaceutical composition according to claim 1, for the treatment of gastric cancer or breast cancer. The pharmaceutical composition for treating gastric cancer according to claim 1, wherein the mutation has any one selected from V842I, R678Q, or a combination thereof. The pharmaceutical composition according to claim 1, for treating breast cancer having any one mutation selected from V777L, L755S, L662Q, or a combination thereof. An effective amount of positiotinib for treating a cancer comprising at least one mutation present in the kinase domain (KD), mucosal membrane domain (JMD), or transmembrane domain (TMD) of human epidermal growth factor receptor 2 (HER2). and an effective amount of a HER2 inhibitor, wherein the mutation is
exon 21 mutation of HER2;
a mutation selected from V777L, L755S, or a combination thereof in the amino acid sequence of HER2;
a mutation comprising a deletion, insertion, or substitution of 1 to 18 nucleotides at amino acid positions 675 to 714 of HER2; and
a mutation comprising a deletion, insertion, or substitution of 1 to 18 nucleotides at amino acid positions 648 to 674 of HER2; Including any one selected from combinations thereof,
The pharmaceutical combination for the treatment of HER2 mutant cancer, wherein the posiotinib and the HER2 inhibitor are administered simultaneously, sequentially or separately. 19. The method of claim 18, wherein the HER2 inhibitor is selected from Trastuzumab, Pertuzumab, Trastuzumab Emtacin, Afatinib, Lapatinib, Neratinib, Trastuzumab deluxecan (DS-8201a), or a combination thereof. a pharmaceutical combination. The pharmaceutical combination according to claim 18, wherein the posiotinib is an oral dosage form and the HER2 inhibitor is an oral dosage form or an injection dosage form. 19. The method of claim 18, wherein pogiotinib is included in a dose of 0.05 to 10 mg/kg/day based on the body weight (kg) of the administered subject, and the HER2 inhibitor is 1 to 250 mg based on the body weight (kg) of the administered subject. A pharmaceutical combination, which is included in a dose of /kg/week. Administering a composition comprising positiotinib of claim 1 to a subject suffering from a cancer comprising at least one mutation appearing in the kinase domain, the mucosal membrane domain, or the transmembrane domain of human epidermal growth factor receptor 2 (HER2). and the mutation is
exon 21 mutation of HER2;
a mutation selected from V777L, L755S, or a combination thereof in the amino acid sequence of HER2;
a mutation comprising a deletion, insertion, or substitution of 1 to 18 nucleotides at amino acid positions 675 to 714 of HER2; and
a mutation comprising a deletion, insertion, or substitution of 1 to 18 nucleotides at amino acid positions 648 to 674 of HER2; A method of treating HER2 mutant cancer, comprising any one selected from a combination thereof. The method of claim 22 , wherein an effective amount of a HER2 inhibitor is further administered to treat the cancer. 24. The method of claim 23, wherein positiotinib and the HER2 inhibitor are administered simultaneously, sequentially or separately. 24. The method of claim 23, wherein the HER2 inhibitor is selected from trastuzumab, pertuzumab, trastuzumab emtacin, afatinib, lapatinib, neratinib, trastuzumab deluxtecan (DS-8201a), or a combination thereof. Any one method of treating cancer. 23. The method of claim 22, wherein the subject analyzes a genomic sample from the patient,
exon 21 mutation of HER2;
a mutation selected from V777L, L755S, or a combination thereof in the amino acid sequence of HER2;
a mutation comprising a deletion, insertion, or substitution of 1 to 18 nucleotides at amino acid positions 675 to 714 of HER2; and
a mutation comprising a deletion, insertion, or substitution of 1 to 18 nucleotides at amino acid positions 648 to 674 of HER2; A method of treating cancer, wherein the subject is identified as comprising any one selected from combinations thereof. The method of claim 26 , wherein the genomic sample is isolated from fluid, blood, urine, normal tissue, or tumor tissue. 23. The method of claim 22, wherein the presence of the mutation is determined by nucleic acid sequencing or PCR analysis.

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