WO2024058289A1 - Pharmaceutical composition, for prevention or treatment of neuroendocrine neoplasm, containing antibody that specifically binds to pd-1 as active ingredient - Google Patents

Abstract

An antibody of the present invention that specifically binds to PD-1, or an antigen-binding fragment of the antibody, can inhibit the activity of PD-1, for example by binding to PD-1 and inhibiting the interaction between PD-1 and PD-L1, and is thus useful in developing immunotherapeutic agents for various neuroendocrine neoplasms.

Description

Pharmaceutical composition for preventing or treating neuroendocrine neoplasms comprising an antibody that specifically binds to PD-1 as an active ingredient

The present invention relates to a pharmaceutical composition for the prevention or treatment of neuroendocrine neoplasms comprising an antibody that specifically binds to PD-1 as an active ingredient.

The first generation chemical anticancer drugs and second generation targeted anticancer drugs that are currently widely used have problems such as side effects due to the toxicity of the anticancer drugs and a high risk of drug resistance. Immunoanticancer drugs, called third-generation anticancer drugs that overcome these problems, act on the signaling pathway of immune cells to activate immune cells and attack cancer cells, producing a therapeutic effect.

PD-1 (also named CD279) is a 55 kD receptor protein related to the CD28/CTLA4 Co-stimulatory/inhibitory receptor family (Blank et al., 2005 Cancer Immunol Immunother 54:307- 314).

The gene and cDNA encoding PD-1 were cloned to examine its characteristics in mice and humans (Ishida et al., 1992 EMBO J 11:3887-3395; Shinohara et al., 1994 Genomics 23:704-706). . Full-length PD-1 contains 288 amino acid residues (NCBI accession number: NP_005009). The extracellular domain consists of 1–167 amino acid residues, and the cytoplasmic C-terminal tail contains residues 191–288, which contain two hypothetical immuno-regulatory motifs, the immunoreceptor tyrosine-based inhibition motif (ITIM; Vivier et al. ., 1997 Immunol Today 18:286-291) and an immunoreceptor tyrosine switch motif (ITSM; Chemnitz et al., 2004 J Immunol 173:945-954).

So far, two sequence-related ligands PD-L1 (B7-H1) and PD-L2 (B7-DC) specifically interact with PD-1 to induce intracellular signaling and induce CD3- and CD28-mediated T-cell signaling. It has been found to inhibit cell activation (Riley, 2009 Immunol Rev 229:114-125), ultimately regulating T-cell activity, such as secretion of other growth factors and cytokines, as well as cell growth, IL-2, and IFN. -It reduces secretion of γ.

Expression of PD-1 is frequently confirmed in immune cells such as T-cells, B-cells, monocytes, and natural killer (NK) cells, and is rarely expressed in other human tissues such as muscle, epithelium, and nervous tissue. It doesn't work. Additionally, high levels of PD-1 expression are often associated with the activation of immune cells. For example, when the human T-cell line Jurkat is activated by Phytohaemagglutinin (PHA) or phorbol ester (12-O-tetradecanoylphorbol-13-acetate or TPA), the expression of PD-1 is upregulated as shown in Western blot. It has been adjusted. Upon stimulation with anti-CD3 antibodies, the same phenomenon was observed in stimulated mouse T- and B-lymphocytes and primary human CD4+ T cells. Effector T cells are stimulated by increasing PD-1 expression, and activated effector T cells are depleted and redirected toward reduced immune activity. Therefore, PD-1-mediated inhibitory signals are known to play an important role in immune tolerance.

Neuroendocrine neoplasms (NEN) is a general term for tumors originating from gastrointestinal neuroendocrine cells, previously called carcinoid tumors. Neuroendocrine tumors have been called carcinoid tumors because the characteristic shape and arrangement of tumor cells are similar to those of typical carcinomas, and are classified into atypical carcinoids and malignant carcinoids depending on their biological characteristics. However, the suffix ‘tumor’ was applied to a benign disease and did not fit the biological characteristics of neuroendocrine tumors. Therefore, after 2010, WHO adopted the name Neoplasm instead of the name Tumor. Neuroendocrine neoplasms are classified into well differentiated neuroendocrine tumor, well differentiated neuroendocrine carcinoma, and poorly differentiated neuroendocrine carcinoma, and are classified into grades and It is classified according to location.

In particular, as a result of a clinical trial on NETs of Pembrolizumab, which was confirmed to have the best treatment effect for solid tumors, the ORR (objective response rate) was confirmed to be at the level of 3.7%, showing a low treatment effect (Strosberg J. et al., Clin Cancer Res. 2020;26(9):2124-2130).

[Prior art literature]

[Non-patent literature]

Strosberg J. et al., Clin Cancer Res. 2020;26(9):2124-2130.

Under this technical background, the present inventors confirmed that an antibody that specifically binds to PD-1 is particularly effective in treating neuroendocrine neoplasms, and completed the present invention.

One aspect of the present invention provides a pharmaceutical composition for the prevention or treatment of neuroendocrine neoplasms, comprising an antibody that specifically binds to PD-1 or an antigen-binding fragment thereof as an active ingredient.

Another aspect of the present invention provides a method for preventing or treating neuroendocrine neoplasms, comprising administering to a subject an antibody or antigen-binding fragment thereof that specifically binds to PD-1.

The antibody or antigen-binding fragment thereof that specifically binds to PD-1 of the present invention can bind to PD-1 and inhibit the activity of PD-1, such as inhibiting the interaction between PD-1 and PD-L1. It is useful in the development of immunotherapeutics for various neuroendocrine neoplasms.

Figure 1 is a graph showing C _max for each patient group.

Figure 2 is a graph showing AUC _ss,6wk for each patient group.

One aspect of the present invention is a pharmaceutical for the prevention or treatment of neuroendocrine neoplasm (NEN) comprising an antibody or antigen-binding fragment thereof that specifically binds to PD-1 (Programmed Cell Death 1) as an active ingredient. A composition is provided.

As used herein, the term “neuroendocrine neoplasm” collectively refers to tumors arising from epithelial cells that have both neurological and endocrine functions. The neuroendocrine neoplasm is also called a carcinoid tumor or carcinoid tumor.

The neuroendocrine neoplasms can be classified according to the degree of cell differentiation. Specifically, neuroendocrine neoplasms are classified according to histologic features (size, lymphatic invasion, mitotic count, Ki-67 labeling index, adjacent organ invasion, presence of metastases, and presence of hormone production). Specifically, they can be classified using the degree of differentiation, mitotic rate (number of mitoses/2 mm2), and Ki-67 index. For classification and definitions of neuroendocrine neoplasms, refer to the full text of the 2019 WHO classification (2019 World Health Organization (WHO) classification).

The neuroendocrine neoplasms can be classified according to the degree of differentiation into well differentiated neuroendocrine tumor, well differentiated neuroendocrine carcinoma, and poorly differentiated neuroendocrine carcinoma. .

At this time, the highly differentiated neuroendocrine tumor is neuroendocrine tumor (NET); Well-differentiated neuroendocrine carcinoma refers to neuroendocrine carcinoma; And poorly differentiated neuroendocrine carcinoma may mean MiNEN (Mixed neuroendocrine-non-neuroendocrine tumor).

In one embodiment, the neuroendocrine neoplasms include neuroendocrine tumor (NET), neuroendocrine carcinoma (NEC), mixed neuroendocrine-non-neuroendocrine tumor (MiNEN), and adenocarcinoma ex-goblet cell (AGCC). It may be one or more selected from the group consisting of carcinoid).

The neuroendocrine neoplasms can be classified as G1 (Grade 1), G2 (Grade 2), or G3 (Grade 3) depending on tumor grade.

In one embodiment, the neuroendocrine tumor (NET) may be classified as G1 NET, G2 NET, or G3 NET.

In one embodiment, the AGCC may be classified as G1 AGCC, G2 AGCC, or G3 AGCC.

In one embodiment, the neuroendocrine neoplasm may be a NET or NEC of G3.

In one embodiment, the neuroendocrine carcinoma (NEC) may be small cell neuroendocrine carcinoma (SCNEC) or large cell neuroendocrine carcinoma (LCNEC) depending on the type of cell.

The neuroendocrine neoplasms can be classified according to the location of onset. Specifically, the site of occurrence of the neuroendocrine neoplasm may be one or more selected from the group consisting of the pituitary gland, thyroid gland, parathyroid gland, thymus, lung, gastrointestinal tract, peripheral nervous system, pancreas, skin, reproductive system, adrenal gland, urinary system, and breast. It is not limited to this.

In one embodiment, the neuroendocrine neoplasms include: Neuroendocrine tumor of the anterior pituitary, Neuroendocrine thyroid tumors, Parathyroid tumors, Thymus carcinoid tumors, Mediastinal carcinoid tumors, Pulmonary neuroendocrine tumors, Gastroenteropancreatic neuroendocrine tumors (GEP-NET), Peripheral nervous system tumors, Pheochromocytoma , Merkel cell carcinoma of skin, trabecular cancer, and urinary tract carcinoid tumor.

As used herein, the term “programmed cell death protein 1 (PD-1)” is a signal transduction protein that plays a role in regulating the activation and function of T cells. PD-1 expressed on T cells binds to PD-L1, a ligand expressed on cancer cells within the tumor microenvironment, which activates the PD-1 signaling pathway and ultimately leads to inactivation of T cells.

As used herein, the term “antibody that specifically binds to PD-1” refers to an antibody that can bind to PD-1, and may be used interchangeably with “anti-PD-1 antibody” herein. . In particular, the form of the antibody may be a whole antibody or antigen binding fragments.

As used herein, the term "antibody" refers to an immunoglobulin (Ig) molecule that reacts immunologically with a specific antigen, and refers to a protein molecule that acts as a receptor that specifically recognizes the antigen, and refers to the entire ( It may be a whole) antibody or antigen binding fragment.

Specifically, the antibodies or antigen-binding fragments thereof that specifically bind to PD-1 include monoclonal antibodies, polyclonal antibodies, single domain antibodies, single chain antibodies, and polyclonal antibodies. Multispecific antibody, human antibody, humanized antibody, chimeric antibody, intrabody, Fv, scFv, Fv linked by disulfide bond (di-scFv), Fab fragment, F( ab') ₂ fragment and any of the above epitope binding fragments, but is not limited thereto. In one embodiment, the antibody may be a monoclonal antibody or a polyclonal antibody.

The heavy and light chains of immunoglobulins may each include a constant region and a variable region.

As used herein, the term "heavy chain" refers to a full-length heavy chain comprising a variable region (VH) and three constant regions, CH1, CH2, and CH3, sufficient to confer specificity for an antigen and its antigen. It is meant to include all combined fragments.

As used herein, the term "light chain" refers to both a full-length light chain and an antigen-binding fragment thereof comprising a variable region (VL) and a constant region (CL) sufficient to confer specificity for an antigen. It means.

As used herein, the term “CDR (Complementarity determining region)” refers to the complementarity determining region that is part of the heavy chain variable region (VH) and light chain variable region (VL) of an antibody.

The light and heavy chain variable regions of the immunoglobulin include three hypervariable regions called complementarity determining regions (CDRs) and four framework regions (FRs). The CDR mainly functions to bind to the epitope of the antigen. The CDRs of each chain are typically called CDR1, CDR2, and CDR3 sequentially starting from the N-terminus, and are identified by the chain on which the specific CDR is located. The FRs of each chain are typically called FR1, FR2, FR3 or FR4 sequentially, starting from the N-terminus, and are identified by the chain on which the particular FR is located.

In one embodiment, the antibody or antigen-binding fragment thereof that specifically binds to PD-1 has a heavy chain variable region comprising the heavy chain CDR1 of SEQ ID NO: 1, the heavy chain CDR2 of SEQ ID NO: 2, and the heavy chain CDR3 of SEQ ID NO: 3. ; and a light chain variable region including light chain CDR1 of SEQ ID NO: 4, light chain CDR2 of SEQ ID NO: 5, and light chain CDR3 of SEQ ID NO: 6.

The heavy chain variable region of the antibody consists of the amino acid sequence of SEQ ID NO: 7 and about 90% or more, about 91% or more, about 92% or more, about 93% or more, about 94% or more, about 95% or more, about 96% or more, about It may comprise or consist of an amino acid sequence having a sequence identity of at least 97%, at least about 98%, at least about 99%, or 100%. In addition, the light chain variable region of the antibody is about 90% or more, about 91% or more, about 92% or more, about 93% or more, about 94% or more, about 95% or more, and about 96% or more of the amino acid sequence of SEQ ID NO: 8. , may comprise or consist of an amino acid sequence having a sequence identity of at least about 97%, at least about 98%, at least about 99%, or 100%.

In one embodiment, the heavy chain variable region consists of the amino acid sequence of SEQ ID NO: 7; The light chain variable region may consist of the amino acid sequence of SEQ ID NO: 8.

The heavy chain of the antibody is about 90% or more, about 91% or more, about 92% or more, about 93% or more, about 94% or more, about 95% or more, about 96% or more, about 97% or more of the amino acid sequence of SEQ ID NO: 9. It may include or consist of an amino acid sequence having sequence identity of at least about 98%, at least about 99%, or 100%. In addition, the light chain of the antibody is about 90% or more, about 91% or more, about 92% or more, about 93% or more, about 94% or more, about 95% or more, about 96% or more of the amino acid sequence of SEQ ID NO. It may comprise or consist of an amino acid sequence having a sequence identity of at least 97%, at least about 98%, at least about 99%, or 100%.

In one embodiment, the heavy chain consists of the amino acid sequence of SEQ ID NO: 9; The light chain may consist of the amino acid sequence of SEQ ID NO: 10. In this specification, the antibody may be referred to as YBL-006.

Immunoglobulin heavy chain constant regions (CH) exhibit different amino acid compositions and sequences and therefore possess different types of antigenicity. Therefore, immunoglobulins can be divided into five categories and referred to as immunoglobulin isotypes, namely IgM, IgD, IgG, IgA and IgE. The corresponding heavy chains are μ chain, δ chain, γ chain, α chain and ε chain, respectively. Additionally, depending on the amino acid composition of the hinge region and the number and location of heavy chain disulfide bonds, the same type of immunoglobulin can be classified into different subtypes. For example, IgG can be classified as IgG1, IgG2, IgG3, and IgG4. Light chains can be classified as κ or λ chains depending on their different constant regions. Each of the five types of IgG can have either a κ or λ chain.

When the PD-1-specific antibody of the present invention includes a constant region, it may include a constant region derived from IgG, IgA, IgD, IgE, IgM, or a partial hybrid thereof.

In one embodiment, the antibody may include a heavy chain constant region of SEQ ID NO: 11, SEQ ID NO: 14, and/or SEQ ID NO: 15, or a light chain constant region of SEQ ID NO: 12.

In one embodiment, the antibody may include the hinge of SEQ ID NO: 13.

As used herein, the term "steady state (ss)" refers to a state in which the drug's entry rate into the body and the elimination rate are equal and are in equilibrium due to repeated administration of the drug. As used herein, AUC and C _max are pharmacokinetic measures of systemic exposure to a drug (e.g. YBL-006) following administration of the drug in humans. The AUC and C _max may be driving factors of drug efficacy.

As used herein, the term “AUC” refers to the area under the curve of a graph showing blood concentration and time of an administered drug.

As used herein, the term “C _max ” refers to the maximum or highest drug concentration observed after drug administration. As used herein, the term “AUC _ss,6wk ” refers to the AUC observed at steady state approximately 4 to 6 weeks after the first drug administration.

In addition, “treatment of neuroendocrine neoplasms” means inhibiting the growth of cells or tissues of neoplasms arising in the neuroendocrine system or preventing the occurrence of such neoplasms, compared to when treated or not treated. The concept also includes reducing cell growth and metastasis and reducing drug resistance to increase the therapeutic effect. The “prevention” refers to all actions that inhibit the development of neoplasms or delay their onset by administering the pharmaceutical composition.

In the pharmaceutical composition for preventing or treating neuroendocrine neoplasms of the present invention, as long as the antibody or antigen-binding fragment thereof can exhibit activity, it may be included in any amount (effective amount) depending on the use, formulation, purpose of formulation, etc. Here, “effective amount” refers to the amount of active ingredient that can induce a therapeutic effect. Such effective amounts can be determined experimentally within the scope of the ordinary ability of those skilled in the art. The pharmaceutical composition of the present invention contains the antibody or antigen-binding fragment thereof as an active ingredient in an amount of about 0.1% to about 90% by weight, specifically about 0.5% by weight to about 75% by weight, more specifically about 0.1% by weight to about 90% by weight, based on the total weight of the composition. It may contain from 1% by weight to about 50% by weight.

In one embodiment, the pharmaceutical composition may be administered to an individual in a total amount of about 100 mg to about 500 mg of the antibody or antigen-binding fragment thereof every 1 day to 4 weeks. Specifically, the pharmaceutical composition containing the antibody or antigen-binding fragment thereof is effective for 1 day to 4 weeks, 2 days to 4 weeks, 3 days to 4 weeks, 4 days to 4 weeks, 5 days to 4 weeks, and 6 days to 4 days. weeks, 1 to 4 weeks, 2 to 4 weeks, 2 to 3 weeks, 3 to 4 weeks, 1 day to 3 weeks, 1 day to 2 weeks, 1 day to 1 week, 1 day to 6 days, It may be administered every 1 to 5 days, 1 to 4 days, 1 to 3 days, or every 1 to 2 days. The antibody or antigen-binding fragment thereof is administered in an amount of about 100 mg to about 450 mg, about 100 mg to about 400 mg, about 100 mg to about 350 mg, about 100 mg to about 320 mg, about 100 mg to about 300 mg, about 150 mg. mg to about 500 mg, about 150 mg to about 450 mg, about 150 mg to about 400 mg, about 150 mg to about 350 mg, about 180 mg to about 320 mg, about 200 mg to about 500 mg, about 200 mg to It may be administered in an amount of about 400 mg, or about 200 mg to about 300 mg. Preferably, it can be administered in an amount of about 200 mg to about 300 mg every 2 to 3 weeks.

The pharmaceutical composition of the present invention may contain a conventional, non-toxic pharmaceutically acceptable carrier that is formulated into a preparation according to a conventional method.

The pharmaceutically acceptable carrier may be any carrier that is a non-toxic material suitable for delivery to a patient. Distilled water, alcohol, fats, waxes and inert solids may be included as carriers. Pharmacologically acceptable adjuvants (buffers, dispersants) may also be included in the pharmacological composition.

As used herein, the term “pharmaceutically acceptable carrier” refers to a carrier or diluent that does not irritate living organisms and does not inhibit the biological activity and properties of the administered compound. Acceptable pharmaceutical carriers in compositions formulated as liquid solutions include those that are sterile and biocompatible, such as saline solution, sterile water, Ringer's solution, buffered saline solution, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and One or more of these ingredients can be mixed and used, and other common additives such as sweeteners, solubilizers, wetting agents, emulsifiers, isotonic agents, absorbents, antioxidants, preservatives, lubricants, fillers, buffers, and bacteriostatic agents are added as needed. can do.

The compositions of the present invention can be prepared in a variety of formulations for parenteral administration (e.g., intramuscular, intravenous, or subcutaneous injection). When the pharmaceutical composition of the present invention is prepared as a parenteral formulation, it can be formulated in the form of injections, transdermal administration, nasal inhalation, and suppositories along with a suitable carrier according to methods known in the art. Injectable preparations include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate. As a base for suppositories, Withepsol, Macrogol, Tween 61, cacao, laurin, glycerogelatin, etc. can be used. Meanwhile, injectables may contain conventional additives such as solubilizers, isotonic agents, suspending agents, emulsifiers, stabilizers, preservatives, etc.

Regarding the formulation of pharmaceutical compositions, it is known in the art, and specifically, references can be made to the literature [Remington's Pharmaceutical Sciences (19th ed., 1995)]. The above documents are considered part of this specification.

The antibody, antigen-binding fragment thereof, or composition containing them of the present invention may be administered to a patient in a therapeutically effective or pharmaceutically effective amount.

The term “administration” used herein means introducing a predetermined substance into an individual by an appropriate method, and the composition can be administered through any general route as long as it can reach the target tissue. The route of administration may include, but is not limited to, intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, topical administration, intranasal administration, and intrarectal administration.

Here, “therapeutically effective amount” or “pharmaceutically effective amount” refers to the amount of a compound or composition effective in preventing or treating the target disease, which is sufficient to treat the disease with a reasonable benefit/risk ratio applicable to medical treatment. It refers to an amount that does not cause side effects. The level of the effective amount is determined by factors including the patient's health status, type and severity of the disease, activity of the drug, sensitivity to the drug, administration method, administration time, administration route and excretion rate, treatment period, combination or concurrent use of drugs, and It may be determined based on other factors well known in the medical field. In one embodiment, a therapeutically effective amount refers to an amount of drug that is effective in treating cancer.

Specifically, the effective amount of the antibody or antigen-binding fragment thereof of the present invention may vary depending on the patient's age, gender, and weight, but generally, the antibody or antigen-binding fragment thereof is administered in an amount of about 100 mg to about every 1 day to 4 weeks. Administration may be repeated to achieve a total dose of 500 mg. However, since it may increase or decrease depending on the route of administration, severity of disease, gender, weight, age, etc., the scope of the present invention is not limited thereto.

The term "individual" refers to an object to which the composition of the present invention can be applied (prescribed), and may be a mammal such as a monkey, dog, cat, rat, mouse, or other livestock, including humans. Preferably, it may be a human, but is not limited thereto.

The antibody, antigen-binding fragment thereof, or pharmaceutical composition containing the same of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or multiple times. there is. Additionally, it may be formulated in the form of a combination preparation with other therapeutic agents. At this time, the other therapeutic agent may include any compound or natural extract whose safety has already been verified and which is known to have therapeutic activity or anti-cancer activity for neuroendocrine neoplasms in order to increase or reinforce therapeutic activity.

Taking all of the above factors into consideration, it is important to administer an amount that can achieve maximum effect with minimal or no side effects, and this can be easily determined by a person skilled in the art.

Another aspect of the present invention provides the use of the antibody or antigen-binding fragment thereof that specifically binds to PD-1 of the present invention for preparing a medicament for the prevention or treatment of neuroendocrine neoplasms.

PD-1, antibodies, antigen-binding fragments, neuroendocrine neoplasms, prevention and treatment are the same as described above.

Another aspect of the present invention provides the use of the antibody or antigen-binding fragment thereof that specifically binds to PD-1 of the present invention for the prevention or treatment of neuroendocrine neoplasms.

The antibodies and antigen-binding fragments thereof, neuroendocrine neoplasms, prevention and treatment are the same as described above.

Another aspect of the present invention is the prevention or treatment of neuroendocrine neoplasms, comprising administering an antibody or antigen-binding fragment thereof that specifically binds to PD-1, or a pharmaceutical composition containing the same, to an individual in need thereof. Provides a method.

PD-1, antibodies, antigen-binding fragments, pharmaceutical compositions, neuroendocrine neoplasms, administration, treatment and prevention are the same as described above.

The subject may be a mammal, preferably a human. Additionally, the individual may be a patient or have a high risk of suffering from neuroendocrine neoplasm.

Another aspect of the present invention provides an antibody or antigen-binding fragment thereof that specifically binds to PD-1 for use in the prevention or treatment of neuroendocrine neoplasm (NEN).

PD-1, antibodies, antigen-binding fragments, pharmaceutical compositions, neuroendocrine neoplasms, treatment and prevention are the same as described above.

Hereinafter, the present invention will be explained in detail by the following examples. However, the following examples are only for illustrating the present invention, and the present invention is not limited thereto.

Example 1. Preparation of YBL-006

YBL-006 is an antibody containing the amino acid sequence shown in Table 1 below. For specific details about YBL-006, refer to the full text of WO 2018-026248.

Specifically, YBL-006 was produced by preparing a vector containing DNA or RNA encoding the amino acid sequence shown in Table 1, then transfecting it into host cells and expressing it. Afterwards, the purification process was performed and completed. The method for producing antibodies is not limited to this, and various methods known to those skilled in the art can be used. For examples of methods for producing antibodies, see the full text of Korean Patent Nos. 10-1093717, 10-2250234 or WO 2018-026248.

Table 1 below shows the heavy chain, light chain, heavy chain CDR (HCDR), light chain CDR (LCDR), and heavy chain variable domain (VH) of YBL-006. ), light chain variable domain (VL), heavy chain constant domain (CH), and light chain constant domain (CL).

	amino acid sequence	sequence number
HCDR1	GFTFLRYA	One
HCDR2	ISYDGRYK	2
HCDR3	TTTTFDS	3
LCDR1	QSLLDSEDGNTY	4
LCDR2	TLS	5
LCDR3	MQRRRDFPFT	6
VH	QVQLVESGGGVVQPGRSLRLSCAASGFTFLRYAMHWVRQAPGKGLEWVAVISYDGRYKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCTTTTFDSWGQGTLVTVSS	7
V.L.	DIVMTQTPLSLPVTPGEAASISCRSSQSLLDSEDGNTYLDWYLQKPGQSPQLLIYTLSHRASGVPDRFSGSGSGTDFTLEISRVEAEDVGVYYCMQRRDFPFTFGQGTKVDIK	8
heavy chain	QVQLVESGGGVVQPGRSLRLSCAASGFTFLRYAMHWVRQAPGKGLEWVAVISYDGRYKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCTTTTFDSWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTK VDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLLSLSLGK	9
light chain	DIVMTQTPLSLPVTPGEAASISCRSSQSLLDSEDGNTYLDWYLQKPGQSPQLLIYTLSHRASGVPDRFSGSGSGTDFTLEISRVEAEDVGVYYCMQRRDFPFTFGQGTKVDIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKV YACEVTHQGLSSPVTKSFNRGEC	10
CH1	ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRV	11
CL	RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC	12
hinge	ESKYGPPCPPCP	13
CH2	APEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAK	14
CH3	GQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK	15

Example 2. Confirmation of therapeutic effect of YBL-006

2.1. test design

Patients were recruited to confirm the therapeutic effect of YBL-006, and the recruited patients gave written consent to the informed consent form (ICF) approved by the Institutional Review Board (IRB)/Independent Ethics Committee (IEC) and satisfied the recruitment conditions. and consisted of those who had no exclusion conditions. The patient group consisted of a total of 8 people.

The treatment period to confirm the treatment effect of YBL-006 consisted of a 14-day (cohort B1 and B3) or 21-day (cohort B2) cycle. The B1 and B3 cohorts were administered 200 mg every 2 weeks (Q2W), and the B2 cohort was administered 300 mg every 3 weeks (Q3W).

YBL-006 was administered once intravenously on Day 1 of each cycle. Tumor response evaluation was performed every 6 weeks. Specifically, for cohorts B1 and B3, it was conducted at the end of every 3 cycles (i.e., 3rd, 6th, and 9th cycles, etc.), and for cohort B2, it was conducted at the end of every 2 cycles (i.e., 2nd, 4th, and 6th cycles, etc.) did. The duration of treatment for patients who responded to therapy was up to 1 year for both the dose escalation and dose expansion phases.

2.2. Inclusion criteria

Clinical trial participants must meet all of the following criteria:

1. Written informed consent in the informed consent form (ICF) approved by the Institutional Review Board (IRB)/Independent Ethics Committee (IEC) prior to evaluation for this study.

2. Male or female ≥18 years of age (or legal adult, or older) at the time of ICF.

3. Life expectancy more than 3 months

4. Evaluation of whole body performance (ECOG) performance status 0 to 1

5. Archival tumor tissue is available and agrees to provide archived tumor for retrospective biomarker analysis or agrees to perform fresh tumor biopsy during screening. Patients who do not have an existing (archived) tumor specimen and do not wish to undergo a biopsy may be enrolled with prior approval from the sponsor.

7. At least one measurable lesion based on Response Evaluation Criteria in Solid Tumors (RECIST) version 1.1. Previously irradiated lesions may be considered target lesions if they are well defined, measurable, and have objective evidence of increase in size.

8. Central nervous system (CNS) metastases must have no evidence of progressive neurological symptoms and require increased corticosteroid doses to control CNS disease for more than 4 weeks. If the patient requires corticosteroids for the management of CNS disease, the dose should be stable at a low dose (less than 10 mg/day of prednisone or equivalent steroid dose) for at least 2 weeks prior to C1D1.

9. Immunosuppressive administration of systemic drugs such as steroids (prednisone at a dose >10 mg/day or equivalent steroid dose) should be discontinued at least 2 weeks prior to IP administration.

10. Previous surgery requiring general anesthesia must be completed at least 14 days prior to IP administration. Surgery requiring local/epidural anesthesia must be completed at least 72 hours prior to IP administration and the patient must be recovered.

11. Adequate hematological and biological function confirmed by laboratory values such as:

Neutrophils ≥ 1000/μL

Platelets ≥ 75,000/μL

Hemoglobin ≥ 9.0 g/dL (may have received a blood transfusion)

Creatinine ≤ 1.5×Upper Limit of Normal (ULN)

Aspartate aminotransferase (AST) ≤ 2.5×ULN (for liver metastases, AST ≤ 5×ULN)

Alanine aminotransferase (ALT) ≤ 2.5×ULN (for liver metastases, AST ≤ 5×ULN) Bilirubin ≤ 1.5×ULN (for patients with Gilbert's syndrome, total bilirubin < 3.0 mg/dL must be).

12. Women must meet one of the following criteria: postmenopausal for at least 24 consecutive months, surgically unable to become pregnant (i.e., hysterectomy or bilateral oophorectomy), or using reliable contraception. In general, decisions about appropriate contraception should be made through discussion between the investigator and the test subject. Women of childbearing potential (WOCBP) must agree to use reliable contraception during the study period and for at least 120 days after the last dose of IP.

13. Men must agree to use an acceptable method of contraception during the study period and for at least 180 days after the last dose of IP and to avoid sperm donation.

14. For expansion cohorts, enrolled patients must also meet the following selection criteria: Confirmed diagnosis of one of the following tumor types:

a. For all expansion cohorts (B1, B2, B2 preliminary, and B3), histologically confirmed advanced solid tumors (primary CNS tumors) that are unresectable, locally advanced, or metastatic and have shown progression after all standard therapies or are ineligible for standard therapy. except).

b. For the B3 cohort, all patients must meet one of the following:

- Patients must have shown progression after at least one standard systemic therapy and no other available approved/standard therapy has been shown to prolong overall survival (either in a randomized clinical trial against another standard of care or when compared to a historical control group) . Patients who, for medical reasons, cannot receive other standard therapies shown to prolong survival are eligible if they meet other eligibility criteria. or

- Patients with disease for which there is no standard treatment shown to prolong overall survival. The following tumor types are eligible for the B3 cohort:

- Metastatic non-small cell lung cancer (NSCLC): PD-L1 tumor expression determined by IHC without EGFR sensitizing (activating) mutations by PCR and anaplastic lymphoma kinase (ALK) translocation determined by IHC or fluorescence in situ hybridization (FISH). High (≥ 50%) histological or cytological confirmation of stage IV NSCLC (for non-squamous cell carcinoma or NSCLC of unknown subtype). For pathologically confirmed squamous cell carcinoma, only high (≥ 50%) PD-L1 tumor expression is required.

In Korea, registered patients must also meet the following requirements:

· If you have received standard anti-cancer treatment including platinum and have not responded to it

· If platinum-based anticancer treatment was refused or discontinued due to intolerance

· If standard immunotherapy treatment was refused or discontinued due to intolerance

- Locally confirmed unresectable or metastatic MSI-H or dMMR solid tumor confirmed by other appropriate methods, such as PCR or IHC, according to local laboratory criteria:

If MSI-H testing has previously been completed, no additional testing is required. However, if the subject did not undergo this test, additional testing must be completed at a local facility.

- Non-clear cell renal cell carcinoma (RCC), anal squamous cell carcinoma (aSCC), cervical cancer, cutaneous squamous cell carcinoma (cSCC), endometrial cancer, high tumor mutation burden (TMB-H) tumor, penile epithelial tumor ( Histologic confirmation of squamous cell carcinoma or adenocarcinoma), neuroendocrine tumor (any origin, pancreatic or non-pancreatic), or nasopharyngeal carcinoma.

TMB-H: Test registration is possible if TMB ≥ 10 mutations/megabase is confirmed in the local (testing institution) laboratory.

- Histologically or cytologically confirmed recurrent or metastatic head and neck squamous cell carcinoma (HNSCC)

2.3. Exclusion criteria

1. History of severe hypersensitivity to other monoclonal antibodies.

2. Anti-PD-1, anti-PD-L1, anti-PD-L2, anti-cytotoxic T lymphocyte associated protein 4 (anti-CTLA-4) antibodies or other antibodies or drug-specific T cell costimulation or immune checkpoints Previous therapy using the route.

3. Adverse reactions due to chemotherapy, radiotherapy, biological cancer therapy, or tyrosine kinase inhibitor (TKI) therapy within 2 weeks or 5 half-lives (longer) prior to the first IP administration, or cancer treatment administered at least 2 weeks prior ( AE) did not recover to grade 1 or lower on the National Cancer Institute (NCI) Common Terminology Criteria for Adverse Events (CTCAE) version 5 or higher (palliative radiotherapy with a limited irradiation area is permitted up to 14 days before the first dose of YBL-006) .

4. Previous malignancy that was active within the previous 2 years, excluding cured locally treatable cancers such as basal or squamous cell skin cancer, superficial bladder cancer, or cervical carcinoma in situ, or breast or localized prostate cancer.

5. Active infection (viral, bacterial, or fungal) requiring IV antibacterial therapy within 14 days prior to first dose of YBL-006.

6. Chronic hepatitis B (e.g. HBs antigen reactivity), hepatitis C infection, or human immunodeficiency virus (HIV) without sufficient antiviral therapy (previous therapy duration must be at least 3 months).

7. Active interstitial lung disease (ILD) or history of pneumonia requiring oral or IV steroids.

8. Evidence of bleeding constitution.

9. Active or suspected autoimmune disease or documented history of autoimmune disease or syndrome requiring systemic steroids or immunosuppressants, excluding patients with vitiligo or recovered childhood asthma/atopy.

10. Administration of a live attenuated vaccine within 28 days prior to the first IP dose (Note: If enrolled, patients must not receive a live vaccine during this study and for 180 days after the last IP dose). Vaccination with killed vaccines is permitted at any time after consultation with a medical monitor. Once a vaccine becomes available, patients can immediately receive vaccination with the appropriate SARS-CoV-2 vaccine (DNA or mRNA vaccine) at the discretion of the investigator.

11. Known current drug or alcohol abuse.

12. Definite active or latent tuberculosis infection (purified protein derivative [PPD] test not required) presented with: PPD that recently converted to positive; Chest x-ray with clear evidence of active infectious infiltrates.

13. The presence of other conditions that may increase the risks associated with trial participation or may interfere with the interpretation of trial results and, in the opinion of the investigator, make the patient unsuitable for trial participation.

14. Concomitant medical conditions requiring the use of immunosuppressants or immunosuppressive doses of systemic corticosteroids (dose > 10 mg/day prednisone or equivalent steroid dose). Inhaled or intranasal corticosteroids (with minimal systemic absorption) can be continued if the patient is on a stable dose. Non-absorbable intra-articular steroid injections are permitted.

15. Use of other investigational therapies within 28 days prior to IP administration.

16. Minor surgical procedure (e.g., central venous access port insertion) ≤ 5 days prior to first dose of IP or major surgical procedure not related to the study ≤ 21 days; In all cases, the patient must be sufficiently recovered and stable prior to administration of therapy.

17. Mental illness or social circumstances that impede test compliance.

18. Congestive heart failure grade 3 to 4 according to the New York Heart Association (NYHA) classification Clinically significant uncontrolled cardiovascular disease, such as myocardial infarction or unstable angina within the previous 6 months, which may cause uncontrolled hypertension or QT prolongation Patients with clinically significant uncontrolled arrhythmias, such as bradyarrhythmias (e.g., type 2 second-degree atrioventricular block or third-degree atrioventricular block).

19. Patients with a corrected QT interval (QTcF) > 470 msec using the Fridericia formula. Patients with a history of prolonged QT syndrome or Torsades de pointes. Patients with a family history of prolonged QT syndrome

Example 3. Pharmacokinetic (PK) analysis

According to Example 2, the blood concentration of YBL-006 administered to the patient group was investigated, and the results and NONMEM® (Boeckmann, A. J., Sheiner, L. B., Beal, S. L. (2020). NONMEM Users Guide - Part V. ICON plc., version 7.5.1) was used to conduct pharmacokinetic analysis. The results are as shown in Table 2 below. For each indicator, refer to the explanation of the symbol below.

Cohort	B1	B2	B3
Dose	200mg Q2W	300mg Q3W	200mg Q2W
C max,est (mg/L)	59.02 ±15.60	68.37 ±19.87	53.87 ±10.22
AUC τ,est (mgh/L)	12549 ±4903	15821 ±5117	11571 ±2369
AUC ss,6wk (mgh/L)	38703 ±8123	39839 ±8956	37234 ±6980

As shown in Table 2 and Figures 1 and 2, the maximum concentration was the group administered 300 mg every 3 weeks (300mg Q3W) (Cohort B2) and the group administered 200 mg every 2 weeks (200mg Q2W) (Cohort It was measured slightly higher than B1, B3). This is within the error range of cohorts B1 and B3, and there was no significant difference even when comparing the C _max.est of each experimental group divided by the administered dose (C _max,est /Dose).

In addition, referring to AUC _ss,6wk , the drug concentration maintained in the body until 6 weeks after administration is the group administered 200 mg every 2 weeks (200mg Q2W) and the group administered 300 mg every 3 weeks (300mg Q3W). It was confirmed that this was at a similar level.

Example 4. Objective response

The objective response results of cohorts B1 to B3 are shown in Table 3 below. Response evaluation was based on Response Evaluation Criteria in Solid Tumors (RECIST) version 1.1 (Eisenhauer EA et al. Eur J Cancer. 2009 Jan;45(2):228-247).

	200mg Q2W	300mg Q3W
CR+PR	8	One
SD	20	6
P.D.	14	4
Sum	42	11

* p-value = 0.8162

* Fisher's Exact Test for count data

Through the results of Example 3 and Example 4, it was confirmed that there was no significant difference between 200mg Q2W and 300mg Q3W in the administration method of YBL-006 for cancer treatment.

Example 5. NET treatment effect

The treatment effect on patients with NET disease was confirmed. Response evaluation was based on Response Evaluation Criteria in Solid Tumors (RECIST) version 1.1 (Eisenhauer EA et al. Eur J Cancer. 2009 Jan;45(2):228-247).

In the table below, SUM means the total, ORR means the objective response rate, and DCR means the disease control rate. ORR was determined as the ratio of patients with complete response (CR) and partial response (PR) compared to the total number of patients, and DCR was determined as the ratio of patients with complete response (CR), partial response (PR), and stable lesion (SD) among the total number of patients. Judgment was made.

	All Grades	NET Grade 1 or 2	NET Grade 3 or NEC
CR	0	0	0
PR	2	0	2
SD	4	One	3
P.D.	3	One	2
SUM	9	2	7
ORR(%)	22.2%	0.0%	28.6%
DCR(%)	66.7%	50.0%	71.4%

The ORR of the NET patient group was 22.2% and the DCR was 66.7%, confirming that YBL-006 has a therapeutic effect on NET. In particular, for Grade 3 patients, the ORR was 28.6% and the DCR was 71.4%. The two patients in whom partial remission was observed were patients with duodenal NEC (large cell) and ampulla of vater NEC, and both were confirmed to have NEC disease of the digestive system.

[Explanation of symbols]

AUC _τ,est : Estimated area under the concentration-time curve, for dosing interval tau

AUC _ss,6wk : Area under the concentration-time curve at steady state, estimated for a period of 6 weeks

CDR: Complementarity determining region

CH: Heavy chain constant region

CI: Confidence interval

CL: Light chain constant region

CL: Clearance

C _max,est : Estimated maximum concentration of drug

CR: Complete response

HC: Heavy chain

IgG4: Immunoglobulin G4

LC: Light chain

NEC: Neuroendocrine carcinoma

NEN: Neuroendocrine neoplasm

NET: Neuroendocrine tumor

ORR: Objective response rate

PD: Progressive disease

PD-1: Programmed cell death-1

PK: Pharmacokinetics

PR: Partial response

Q2W: Every two weeks

Q3W: Every three weeks

SD: Stable disease

SD: Standard deviation

Vc: Central volume of distribution in the central compartment

Vp: Peripheral volume of distribution in peripheral compartment

Vss: Volume of distribution at steady state

VH: Heavy chain variable region

VL: Light chain variable region

[National research and development project that supported this invention]

This study was conducted with the support of the National New Drug Development Project of the National New Drug Development Project with financial resources from the Ministry of Science and ICT, the Ministry of Trade, Industry and Energy, and the Ministry of Health and Welfare (project identification number: HN21C1391000021).

Claims (14)

1. A heavy chain variable region comprising the heavy chain CDR1 of SEQ ID NO: 1, the heavy chain CDR2 of SEQ ID NO: 2, and the heavy chain CDR3 of SEQ ID NO: 3; and a neuroendocrine neoplasm comprising an antibody or antigen-binding fragment thereof containing a light chain variable region including light chain CDR1 of SEQ ID NO: 4, light chain CDR2 of SEQ ID NO: 5, and light chain CDR3 of SEQ ID NO: 6 as an active ingredient. , NEN) pharmaceutical composition for prevention or treatment.
2. According to paragraph 1,

   A pharmaceutical composition for the prevention or treatment of neuroendocrine neoplasms, wherein the antibody is a monoclonal antibody or polyclonal antibody.
3. According to paragraph 1,

   The antibody or antigen-binding fragment thereof is

   A heavy chain variable region comprising a sequence having more than 90% sequence identity with the amino acid sequence of SEQ ID NO: 7; and

   A pharmaceutical composition for the prevention or treatment of neuroendocrine neoplasms, comprising a light chain variable region comprising a sequence having more than 90% sequence identity with the amino acid sequence of SEQ ID NO: 8.
4. According to paragraph 1,

   The neuroendocrine neoplasms are from the group consisting of Neuroendocrine tumor (NET), Neuroendocrine Carcinoma (NEC), Mixed neuroendocrine-non-neuroendocrine tumor (MiNEN), and Adenocarcinoma ex-goblet cell carcinoid (AGCC). A pharmaceutical composition for the prevention or treatment of neuroendocrine neoplasms, comprising one or more selected substances.
5. According to clause 4,

   A pharmaceutical composition for the prevention or treatment of neuroendocrine neoplasms, wherein the neuroendocrine neoplasm is G3 (Grade 3) NET or NEC.
6. According to clause 4,

   A pharmaceutical composition for the prevention or treatment of neuroendocrine neoplasms, wherein the neuroendocrine carcinoma (NEC) is small cell neuroendocrine carcinoma (SCNEC) or large cell neuroendocrine carcinoma (LCNEC).
7. According to clause 4,

   The AGCC is a pharmaceutical composition for the prevention or treatment of neuroendocrine neoplasms, wherein the AGCC is G1 (Grade 1) AGCC, G2 (Grade 2) AGCC, or G3 (Grade 3) AGCC.
8. According to paragraph 1,

   A pharmaceutical composition for the prevention or treatment of neuroendocrine neoplasms, wherein the neuroendocrine neoplasms are functional or non-functional.
9. According to paragraph 1,

   The site of occurrence of the neuroendocrine neoplasm is one or more selected from the group consisting of the pituitary gland, thyroid gland, parathyroid gland, thymus, lung, gastrointestinal tract, peripheral nervous system, pancreas, skin, genitals, adrenal gland, urinary system and breast. Pharmaceutical composition for prevention or treatment of.
10. According to paragraph 1,

    The neuroendocrine neoplasms include Neuroendocrine tumor of the anterior pituitary, Neuroendocrine thyroid tumors, Parathyroid tumors, Thymus carcinoid tumors, and mediastinal carcinoid tumors. ), Pulmonary neuroendocrine tumors, Gastroenteropancreatic neuroendocrine tumors (GEP-NET), Peripheral nervous system tumors, Pheochromocytoma, Merkel cell carcinoma of the skin ( A pharmaceutical composition for the prevention or treatment of neuroendocrine neoplasms, which is at least one selected from the group consisting of Merkel cell carcinoma of skin, trabecular cancer and urinary tract carcinoid tumor.
11. According to paragraph 1,

    A pharmaceutical composition wherein the antibody or antigen-binding fragment thereof is administered in a dosage of 100 mg to 500 mg every 1 day to 4 weeks.
12. According to clause 11,

    A pharmaceutical composition wherein the antibody or antigen-binding fragment thereof is administered in a dosage of 200 mg to 300 mg every 2 to 3 weeks.
13. A heavy chain variable region comprising the heavy chain CDR1 of SEQ ID NO: 1, the heavy chain CDR2 of SEQ ID NO: 2, and the heavy chain CDR3 of SEQ ID NO: 3; and administering to an individual an antibody or antigen-binding fragment thereof comprising a light chain variable region comprising light chain CDR1 of SEQ ID NO: 4, light chain CDR2 of SEQ ID NO: 5, and light chain CDR3 of SEQ ID NO: 6. Methods for preventing or treating organisms (Neuroendocrine neoplasm, NEN).
14. A heavy chain variable region comprising the heavy chain CDR1 of SEQ ID NO: 1, the heavy chain CDR2 of SEQ ID NO: 2, and the heavy chain CDR3 of SEQ ID NO: 3, for use in the prevention or treatment of neuroendocrine neoplasm (NEN); and an antibody or antigen-binding fragment thereof comprising a light chain variable region comprising light chain CDR1 of SEQ ID NO: 4, light chain CDR2 of SEQ ID NO: 5, and light chain CDR3 of SEQ ID NO: 6.

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