CD38 binding agents and uses thereof

Abstract

In particular, the present invention provides compounds that bind to CD38. In some embodiments, the present invention provides conjugates comprising a CD38 binding moiety. In some embodiments, provided compounds are useful for treating conditions, disorders or diseases associated with CD38.

Description

Technical Field

[0001] The present invention relates to agents for binding peptides to CD38, and peptides that specifically bind to human CD38. Background Art

[0002] CD38 is expressed by various types of cells in humans and has many important functions. In some embodiments, CD38 is associated with various conditions, disorders or diseases.

[0003] CD38, also known as cyclic ADP-ribose hydratase, is a type II transmembrane glycoprotein consisting of a long C-terminal extracellular domain and a short N-terminal intracellular domain.

[0004] CD38 is also a member of a "superfamily" of membrane-bound or soluble enzymes, ADP-ribosyl cyclases, which also include CD157 and Aplysia ADPR. These superfamily enzymes convert NAD to cyclic ADP-ribose or nicotinic acid-adenine dinucleotide phosphate.

[0005] In addition, CD38 has been reported to be involved in the CA2+ signaling pathway and other signaling pathways, such as phospholipase Cγ, ZAP-70, syk, and c-cbl. These reports suggest that CD38 plays an important role as a signaling molecule in the normal development, maturation, and activation of lymphoid cells. Many functions in hematopoietic stem cells are caused by CD38-mediated signaling pathways, including the increase of lymphocytes, the release of cytokines, the development and death of B cells and myeloid cells, and the induction of dendritic cell maturation. In addition, CD38 is known to be involved in cell-cell adhesion.

[0006] Thus, CD38 is an important molecule involved in many biological phenomena. Recently, the association between CD38 and malignancies, especially hematological malignancies, has received much attention. It is well known that those malignancies have increased CD38 expression. In addition to this fact, the first stage pluripotent stem cells do not express CD38 (CD38 - ), CD38 is speculated to be a potential target for antibody therapy of hematological malignancies and chronic lymphocytic leukemia.

[0007] Prior art literature

[0008] Patent Literature

[0009] Patent Document 1: WO2016 / 146639

[0010] It is known that antibodies that specifically bind to CD38 are used to treat cancer. There are many documents describing anti-CD38 antibodies (see Patents 1 and 2). Patent 2 discloses the activity of antibody-dependent cellular cytotoxicity and CDC complement-dependent cytotoxicity. In addition, Patents 3 to 6 report the combined use of anti-CD38 antibodies and cytotoxic compounds such as cytarabine, vincristine cyclophosphamide and melphalan.

[0011] CD38 is also known as a biomarker molecule for determining HIV infection, leukemia, myeloma, solid tumors, diabetes, bone metabolism and other diseases or genetic conditions. In particular, it is well known that CD38 is used as a prognostic indicator for leukemia (Ibrahim, S. et al. (2001) CD38 expression as an important prognostic factor in B-cell chronic lymphocytic leukemia. Blood 98: 181-186) or a marker for multiple myeloma.

[0012] However, antibodies are sometimes unable to reach their targets due to their large size. Since this problem would be a major problem for therapies using conjugates of antibodies and cytotoxic compounds, molecules that are smaller than antibodies but can specifically bind to CD38 are sought. Summary of the invention

[0013] In particular, the present invention provides technologies (e.g., compounds, compositions, methods, etc.) for modulating CD38 activity. In some embodiments, the present invention provides technologies that can bind to CD38 and modulate one or more properties and / or functions of CD38 and / or entities that contain and / or express CD38.

[0014] In particular, in some embodiments, the compounds provided herein can bind to CD38 with high affinity. In some embodiments, the compounds provided are or comprise a peptide portion comprising one or more residues of an amino acid or its analog. In some embodiments, the compounds provided are or comprise (Xaa)y as described herein or a salt thereof. In some embodiments, the compounds provided are or comprise -Xaa as described herein T1 -Xaa T2 -(Xaa)y'-Xaa T3 -Xaa T4 -Xaa T5 - or its salt form. In some embodiments, provided compounds are or contain -Xaa described herein T6 -(Xaa)y'-Xaa T7 -Xaa T8 -XaaT9 -Xaa T10 -Xaa T11 - or its salt form.

[0015] In some embodiments, the provided compounds are much smaller in size than CD38 antibodies and / or can provide many benefits in a variety of uses compared to CD38 antibodies, such as for detection, diagnosis or therapeutic agents or fragments thereof. As will be appreciated by those skilled in the art, in some embodiments, the provided compounds are easier to manufacture than antibodies, particularly for large-scale commercial and / or therapeutic purposes. In some embodiments, the provided compounds can be prepared using synthesis, thus providing higher purity and / or consistency compared to antibodies. In some embodiments, the provided compounds and compositions thereof provide significantly improved stability, bioavailability, delivery properties and other benefits for various uses including therapeutic agents or fragments thereof.

[0016] In some embodiments, provided compounds are conjugates comprising: a first moiety that binds to CD38, such as a peptide moiety described herein; a second moiety that can be used for a variety of purposes, such as detection, diagnosis, treatment, etc.; and optionally a linking moiety that connects the first and second moieties. Various techniques for preparing such conjugates, including chemistry, useful second moieties, and linking moieties, etc., are available in the art and can be utilized in accordance with the present invention. For example, in some embodiments, provided compounds are peptide-drug conjugates (PDCs), which comprise a peptide moiety that binds to CD38, a drug moiety that corresponds to a drug, and optionally a linker that connects the peptide moiety and the drug moiety. In some embodiments, PDCs utilize CD38 binding moieties to target specific targets, such as diseased cells expressing CD38, and are capable of specifically delivering the drug moiety (in other types of conjugates, the second conjugate can be a detection or diagnostic moiety, depending on the desired use). In some embodiments, a drug is a toxin. In some embodiments, a drug is a small molecule inhibitor, such as various therapeutically relevant enzymes or other types of proteins. In some embodiments, a drug is a nucleotide drug. Those skilled in the art will appreciate that many techniques in the field of antibody-drug conjugates (ADCs) can be utilized in accordance with the present invention, in particular. In some embodiments, a PDC is provided by replacing the antibody portion of the ADC with, for example, a CD38 binding moiety as described herein, wherein the drug and / or linker moiety is selected to be optimized, the CD38 binding moiety being, for example, one of the various cyclic peptide moieties described herein.

[0017] In some embodiments, the present invention provides methods for identifying, evaluating, preparing, and using provided compounds and compositions, for example, as described in the Examples.

[0018] The present invention provides techniques (eg, compounds, compositions, methods, etc.) for regulating CD38 activity. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 is an exemplary SPR result for compound 1-1.

[0020] Figure 2 is an exemplary SPR result for compound 1-2. DETAILED DESCRIPTION

[0021] 1. Overview of Certain Embodiments

[0022] The agents provided herein, in particular the various compounds such as those described herein, can bind to CD38 to modulate CD38 function and / or deliver useful agents to targets comprising and / or expressing CD38. In some embodiments, the compounds provided are peptides and / or comprise a peptide portion, in many cases a cyclic peptide portion, that can bind to CD38. In some embodiments, the peptide portion comprises one or more residues of an amino acid or an amino acid analog.

[0023] In some embodiments, an amino acid analog is a compound in which the amino group and / or the carboxylic acid group are independently replaced by an optionally replaced aliphatic or heteroaliphatic moiety. As will be appreciated by those skilled in the art, many amino acid analogs that mimic the structure, properties and / or functions of amino acids have been described in the art and can be used according to the present invention.

[0024] The present invention provides a peptide that binds to CD38, and a method for treating, diagnosing, and determining CD38 using a reagent containing the peptide.

[0025] The first aspect of the present invention relates to a CD38 binding peptide, a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable solvate thereof, wherein the CD38 binding peptide is:

[0026] (1) A polypeptide having an amino acid sequence represented by SEQ ID NO. 1 or 2:

[0027] Ala Arg Ahp Tyr His Asp Gly Val Leu Bph Ahp Asp Cys (SEQ ID NO.1), AlaLeu His MePhe Val Leu Pro Bph Val Trp Val Cys (SEQ ID NO.2);

[0028] (2) A polypeptide having an amino acid sequence represented by SEQ ID NO. 1 or 2, wherein the Ala at the N-terminus is chloroacetylated Ala;

[0029] (3) A polypeptide having an amino acid sequence in which one or more amino acids are deleted, added, substituted or inserted in SEQ ID NO. 1 or 2, and excluding an amino acid sequence having a Cys deletion at the C-terminus of SEQ ID NO. 1 or 2;

[0030] (4) A polypeptide having an amino acid sequence represented by SEQ ID NO. 1 or 2, wherein the Ala at the N-terminus is chloroacetylated Ala, and one or more amino acids are deleted, added, substituted or inserted in the amino acid sequence of SEQ ID NO. 1 or 2, and does not include an amino acid sequence having a Cys deletion at the C-terminus of SEQ ID NO. 1 or 2; or

[0031] (5) A polypeptide, which is any one of (1) to (4) above, wherein the polypeptide has a cyclized structure.

[0032] A preferred embodiment of the present invention is that the CD38 binding peptide has an amino acid sequence represented by one of SEQ ID NOs. 1 to 34.

[0033] A preferred embodiment of the present invention is that the CD38 binding peptide has a cyclized structure.

[0034] The second aspect of the present invention relates to a compound comprising the CD38 binding peptide, its pharmaceutically acceptable salt or its pharmaceutically acceptable solvate as described above, wherein the CD38 binding peptide, its pharmaceutically acceptable salt or its pharmaceutically acceptable solvate further comprises a linking portion.

[0035] A preferred embodiment of the present invention is that the linking moiety comprises polyethylene glycol (PEG).

[0036] The third aspect of the present invention relates to a pharmaceutical composition for treating a state, disorder or disease associated with CD38, comprising: a CD38 binding peptide as described above, a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable solvate thereof; and a pharmaceutically acceptable carrier.

[0037] A preferred embodiment of the present invention is that the state, disorder or disease associated with CD38 is cancer.

[0038] A preferred embodiment of the present invention is that the state, disorder or disease associated with CD38 is leukemia or myeloma.

[0039] A preferred embodiment of the present invention is that the state, disorder or disease associated with CD38 is B-cell non-Hodgkin lymphoma (NHL), multiple myeloma (MM), acute myeloid leukemia (AML), acute lymphocytic leukemia (B-cell ALL) or chronic lymphocytic leukemia (CLL).

[0040] A preferred embodiment of the present invention is that the state, disorder or disease associated with CD38 is multiple myeloma (MM).

[0041] The fourth aspect of the present invention relates to a cancer testing reagent, wherein the reagent comprises the CD38 binding peptide as described above, or a pharmaceutically acceptable salt or a pharmaceutically acceptable solvate thereof.

[0042] 2. Definition

[0043] The compounds of the present invention include those generally described herein and further illustrated by the classes, subclasses and species disclosed herein. As used herein, unless otherwise indicated, the following definitions apply. For the purposes of the present invention, chemical elements are according to the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75 th Ed. In addition, the general principles of organic chemistry are described in Organic Chemistry by Thomas Sorrell, University Science Books, Sausalito: 1999, and March's Advanced Organic Chemistry 5 th Ed., Ed.: Smith, M. B. and March, J., John Wiley & Sons, New York: 2001.

[0044] As used herein, unless the context clearly indicates otherwise, (i) the terms "a" and "an" may be understood to mean "at least one"; (ii) the term "or" may be understood to mean "and / or"; (iii) the terms "comprising", "comprise", "including, whether or not used with "not limited to" and "include, whether or not used with "not limited to") may be understood to cover the listed components or steps, whether shown alone or with one or more other components or steps; (iv) the term "another" may be understood to mean at least one additional / second, or more; (v) the terms "about" and "approximately" may be understood to allow for standard variations, which are understood by those of ordinary skill in the art; (vi) ranges are provided with endpoints included. Unless otherwise indicated, the compounds described herein may be provided and / or used in the form of salts, particularly pharmaceutically acceptable salts.

[0045] Pharmaceutical composition: As used herein, the term "pharmaceutical composition" refers to an active agent formulated with one or more pharmaceutically acceptable carriers. In some embodiments, an active agent is present in a unit dose suitable for administration in a treatment method, wherein the treatment method shows a statistically significant probability of achieving a predetermined therapeutic effect when administered to a relevant population. In some embodiments, the pharmaceutical composition can be specifically formulated for administration in solid or liquid form, including those suitable for the following uses: oral administration, such as drops (aqueous or non-aqueous solutions or suspensions), tablets, such as tablets for buccal, sublingual and systemic absorption, pills, powders, granules, pastes suitable for tongue; injection administration, such as by subcutaneous, intramuscular, intravenous or epidural injection, such as sterile solutions or suspensions or sustained release formulations; topical application, such as in the form of creams, ointments or controlled release patches or sprays for administration to the skin, lungs or mouth; intravaginal or rectal, such as vaginal suppositories, creams or foams; sublingual; eye; transdermal; or nasal, lung and other mucosal surfaces.

[0046] Pharmaceutically acceptable: As used herein, the term "pharmaceutically acceptable" refers to those compounds, materials, compositions and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic response, or other problems or complications, and have a reasonable benefit / risk ratio.

[0047] Pharmaceutically acceptable carrier: As used herein, the term "pharmaceutically acceptable carrier" means a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, or solvent encapsulating material, which is involved in carrying or transporting the subject compound from one organ or part of the body to another. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of pharmaceutically acceptable carrier materials include: sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols such as propylene glycol; polyols such as glycerol, sorbitol, mannitol and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethanol; pH buffer solutions; polyesters, polycarbonates and / or polyanhydrides; and other nontoxic compatible substances used in pharmaceutical formulations.

[0048] Pharmaceutically acceptable salts: As used herein, the term "pharmaceutically acceptable salts" refers to salts of such compounds suitable for use in a pharmaceutical setting, that is, salts suitable for contact with human and lower animal tissues without excessive toxicity, irritation, allergic reactions, etc., within the scope of reasonable medical judgment, and with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, SM Berge, et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 66: 1-19 (1977). In some embodiments, pharmaceutically acceptable salts include but are not limited to non-toxic acid addition salts, which are: salts of amino groups formed with inorganic acids, such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid; salts of amino groups formed with organic acids, such as acetic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid; or amino salts formed by using other methods used in the art, such as ion exchange. In some embodiments, pharmaceutically acceptable salts include, but are not limited to, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxyethanesulfonate, lactobionate, lactate, laurate, dodecylsulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, and the like. In some embodiments, the provided compounds contain one or more acidic groups, and the pharmaceutically acceptable salts are alkali metals, alkaline earth metals, or ammonium salts (e.g., ammonium salts of N(R)3, wherein each R is independently defined and described herein). Representative alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. In some embodiments, the pharmaceutically acceptable salts are sodium salts. In some embodiments, the pharmaceutically acceptable salts are potassium salts. In some embodiments, the pharmaceutically acceptable salts are calcium salts. In some embodiments, the pharmaceutically acceptable salts, when appropriate, include non-toxic ammonium, quaternary ammonium, and ammonia cations formed using, for example, halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, alkyls having 1 to 6 carbon atoms, sulfonates, and aryl sulfonates. In some embodiments, the provided compounds contain more than one acid group.In some embodiments, pharmaceutically acceptable salts or general salts of such compounds contain two or more cations that may be the same or different. In some embodiments, in pharmaceutically acceptable salts (or salts in general), all ionizable hydrogens in acidic groups (e.g., in aqueous solution with a pKa of no greater than about 11, 10, 9, 8, 7, 6, 5, 4, 3, or 2; in some embodiments, no greater than about 7; in some embodiments, no greater than about 6; in some embodiments, no greater than about 5; in some embodiments, no greater than about 4; in some embodiments, no greater than about 3) are replaced by cations.

[0049] Pharmaceutically acceptable salt: As used herein, the term "pharmaceutically acceptable salt" refers to salts of such compounds that are suitable for use in a pharmaceutical setting, that is, salts that are, within the scope of sound medical judgment, suitable for contact with the tissues of humans and lower animals without excessive toxicity, irritation, allergic response, etc., and commensurate with a reasonable benefit / risk ratio.

[0050] Protecting group: As used herein, the term "protecting group" is well known in the art and is included in Protecting Groups in Organic Synthesis, TW Greene and PGM Wuts, 3 rd The present invention is described in detail in the 1999 edition, John Wiley & Sons, the entire contents of which are incorporated herein by reference. It also includes protecting groups particularly suitable for those nucleoside and nucleotide chemistry described in Current Protocols in Nucleic Acid Chemistry, edited by Serge L. Beaucage et al. 06 / 2012, the entire contents of Chapter 2 of which are incorporated herein by reference.

[0051] Subject: As used herein, the term "subject" refers to any organism to which a compound or composition of the invention is administered, for example, for experimental, diagnostic, preventive and / or therapeutic purposes. Typical subjects include animals (e.g., mammals such as mice, rats, rabbits, non-human primates and humans; insects; worms, etc.) and plants. In some embodiments, the subject is a human. In some embodiments, the subject may suffer from and / or be susceptible to a disease, disorder and / or condition.

[0052] Substantially: As used herein, the term "substantially" refers to the qualitative condition of exhibiting a full or nearly full degree or level of a characteristic or property of interest. One of ordinary skill in the art will appreciate that biological and chemical phenomena rarely, if ever, reach completion and / or proceed to completion, or achieve or avoid an absolute result. Thus, the term "substantially" as used herein is intended to capture the potential lack of integrity inherent in many biological and / or chemical phenomena.

[0053] Susceptibility: An individual who is "susceptible" to a disease, disorder, and / or condition is an individual who is at a higher risk of developing the disease, disorder, and / or condition than the general population. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition is susceptible to the disease, disorder, and / or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition may not have been diagnosed with the disease, disorder, and / or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition may show symptoms of the disease, disorder, and / or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition may not show symptoms of the disease, disorder, and / or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition will develop the disease, disorder, and / or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition will not develop the disease, disorder, and / or condition.

[0054] Therapeutic agent: As used herein, the term "therapeutic agent" generally refers to any agent that, when administered to a subject, causes a desired effect (e.g., a desired biological, clinical or pharmacological effect). In some embodiments, an agent is considered a therapeutic agent if it exhibits a statistically significant effect throughout a suitable population. In some embodiments, a suitable population is a population of subjects suffering from and / or susceptible to a disease, disorder and / or state. In some embodiments, a suitable population is a population of model organisms. In some embodiments, a suitable population can be defined by one or more criteria, such as age group, sex, genetic background, pre-existing clinical state, prior treatment. In some embodiments, a therapeutic agent is a substance that, when administered to a subject in an effective amount, can alleviate, improve, mitigate, inhibit, prevent, delay the onset of, reduce the severity of a disease and / or reduce the incidence of one or more symptoms or characteristics of the disease, disorder and / or state in the subject. In some embodiments, a "therapeutic agent" is an agent that has been or needs to be approved by a government agency before it can be sold for human administration. In some embodiments, a "therapeutic agent" is an agent that requires a medical prescription to be administered to a person. In some embodiments, the therapeutic agent is a compound described herein.

[0055] Therapeutically effective amount: As used herein, the term "therapeutically effective amount" means the amount of a substance (e.g., a therapeutic agent, composition, and / or formulation) that, when administered as part of a treatment regimen, causes a desired biological response. In some embodiments, a therapeutically effective amount of a substance is an amount sufficient to treat, diagnose, prevent, and / or delay a disease, disorder, and / or state when administered to a subject suffering from or susceptible to the disease, disorder, and / or state. As will be appreciated by those of ordinary skill in the art, the effective amount of a substance may vary depending on factors such as the desired biological endpoint, the substance to be delivered, the target cell or tissue, and the like. For example, an effective amount of a compound in a formulation for the treatment of a disease, disorder, and / or state can alleviate, improve, mitigate, inhibit, prevent, delay onset, reduce severity, and / or reduce one or more symptoms or characteristics of the disease, disorder, and / or state. In some embodiments, a therapeutically effective amount is administered in a single dose. In some embodiments, multiple unit doses are required to deliver a therapeutically effective amount.

[0056] Treatment: As used herein, the terms "treat," "treatment," or "treating" refer to any method for partially or completely alleviating, ameliorating, alleviating, inhibiting, preventing, delaying the onset of, alleviating the severity of, and / or reducing one or more symptoms or characteristics of the disease, disorder, and / or state. Subjects that do not show signs of the disease, disorder, and / or state may be treated. In some embodiments, subjects that only show early signs of the disease, disorder, and / or state may be treated, for example, with the goal of reducing the risk of developing pathologies associated with the disease, disorder, and / or state.

[0057] Unit dose: As used herein, the expression "unit dose" refers to the amount administered in a single dose and / or in physically discrete units of a pharmaceutical composition. In many embodiments, a unit dose contains a predetermined amount of an active agent. In some embodiments, a unit dose contains an entire single dose of an agent. In some embodiments, more than one unit dose is administered to achieve a total single dose. In some embodiments, multiple unit doses are required or expected to be administered in order to achieve the desired efficacy. A unit dose can be, for example, a volume of a liquid (e.g., an acceptable carrier) containing a predetermined amount of one or more therapeutic agents, a predetermined amount of one or more solid therapeutic agents, a sustained release formulation or drug delivery device containing a predetermined amount of one or more therapeutic agents, etc. It should be understood that a unit dose can be present in a formulation that contains any of a variety of ingredients in addition to the therapeutic agent. For example, as described below, an acceptable carrier (e.g., a pharmaceutically acceptable carrier), a diluent, a stabilizer, a buffer, a preservative, etc. may be included. One skilled in the art will appreciate that in many embodiments, the total appropriate daily dose of a particular therapeutic agent may contain a portion or multiple unit doses and may be determined, for example, by an attending physician within the scope of reasonable medical judgment. In some embodiments, a specific effective dosage level for any particular subject or organism may depend on a variety of factors, including: the disease being treated and the severity of the disease; the activity of the specific active compound being used; the specific composition being used; the age, weight, general health, sex, and diet of the subject; the time of administration and rate of excretion of the specific active compound being used; the duration of treatment; drugs and / or additional therapies used in combination or concomitantly with the specific compound being used; and like factors well known in the medical arts.

[0058] Unsaturated: As used herein, the term "unsaturated" means that a moiety has one or more units of unsaturation.

[0059] Unless otherwise stated, structures described herein are also meant to include all isomeric (e.g., mirror image, diastereomeric, and geometric (or configuration)) forms of the structure, for example, R and S configurations for each asymmetric center, Z and E double bond isomers, and Z and E configuration isomers. Therefore, single stereochemical isomers as well as mirror image, diastereomeric, and geometric (or configuration) mixtures of the compounds of the present invention are within the scope of the present invention. Unless otherwise stated, all tautomers of the compounds are within the scope of the present invention. In addition, unless otherwise stated, structures described herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, a compound having a structure of the present invention comprises a hydrogen replaced by a deuterium or tritium, or an isotopically enriched atom replaced by a deuterium or tritium. 13 C- or 14Compounds having a C-carbon in place of a carbon are within the scope of the present invention. Such compounds are useful, for example, as analytical tools, probes in biological assays, or therapeutic agents as in the present invention.

[0060] 3. Description of exemplary embodiments:

[0061] Amino acid: In the present specification, amino acid means any known protein amino acid that is naturally encoded or found in the genetic code of any organism, or non-protein amino acid that is not naturally encoded or found in the genetic code of any organism. Examples of non-protein amino acids include α, α-disubstituted amino acids (α-methylalanine, etc.), N-alkyl-α-amino acids, N-alkyl-α-D-amino acids, whose main chain structures are different from the natural types. Examples include, but are not limited to, β-amino acids and amino acids having side chain structures different from the side chain structures of the natural types (e.g., norleucine, homohistidine, and hydroxyproline).

[0062] CD38: CD38 (cluster of differentiation 38, also known as cyclic ADP ribose hydrolase) is expressed by various types of cells and performs a variety of functions. CD38 has been reported to be found in many immune cells, such as CD4 + 、CD8 + CD38 is a glycoprotein on the surface of cells such as B lymphocytes, B lymphocytes and natural killer cells. CD38 has also been reported to have other functions, such as cell adhesion, signal transduction and calcium signaling. The preferred CD38 is human CD38.

[0063] CD38 has been implicated in various diseases, disorders and / or conditions, such as HIV infection, leukemia, myeloma, solid tumors, CLL, MM, APL (described below), and the like.

[0064] Daratumumab, an antibody targeting CD38, has been approved for the treatment of multiple myeloma.

[0065] CD38 is also a lineage-independent type II transmembrane glycoprotein that synthesizes and hydrolyzes cyclic adenosine 5'-diphosphate ribose, an intracellular calcium mobilizing messenger. Both the release of soluble protein and the ability of membrane-bound protein to be internalized indicate extracellular and intracellular functions of the protein. This protein has an N-terminal cytoplasmic tail, a transmembrane domain, and a C-terminal extracellular region with four N-glycosylation sites. Crystal structure analysis shows that the functional molecule is a dimer, the central portion of which contains the catalytic site. It is used as a prognostic indicator for patients with chronic lymphocytic leukemia. Alternative splicing produces multiple transcript variants.

[0066] CD38 binding agent: In some embodiments, the agent of the present invention can bind to CD38. In this specification, a peptide that specifically binds to CD38 (also referred to as a CD38 binding peptide) means any peptide that can specifically bind to CD38. Those skilled in the art can confirm it according to conventional methods.

[0067] The agent may be a CD38 binding peptide, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof, wherein the CD38 binding peptide is:

[0068] (1) A polypeptide having an amino acid sequence represented by SEQ ID NO. 1 or 2: Ala Arg Ahp TyrHis Asp Gly Val Leu Bph Ahp Asp Cys (SEQ ID NO. 1), Ala Leu His MePhe Val LeuPro Bph Val Trp Val Cys (SEQ ID NO. 2);

[0069] (2) A polypeptide having an amino acid sequence represented by SEQ ID NO. 1 or 2, wherein the Ala at the N-terminus is chloroacetylated Ala;

[0070] (3) A polypeptide having an amino acid sequence in which one or more amino acids are deleted, added, substituted or inserted in SEQ ID NO. 1 or 2, and excluding an amino acid sequence having a Cys deletion at the C-terminus of SEQ ID NO. 1 or 2;

[0071] (4) A polypeptide having an amino acid sequence represented by SEQ ID NO. 1 or 2, wherein the Ala at the N-terminus is chloroacetylated Ala, and one or more amino acids are deleted, added, substituted or inserted in the amino acid sequence of SEQ ID NO. 1 or 2, and does not include an amino acid sequence having a Cys deletion at the C-terminus of SEQ ID NO. 1 or 2; or

[0072] (5) A polypeptide, which is any one of (1) to (4) above, wherein the polypeptide has a cyclized structure.

[0073] CD38-related diseases: CD38 has been reported to be expressed on immune system cells such as T cells or B cells in healthy humans. In the case of disease, it has been determined that CD38 is expressed in increased amounts or specifically on cells that are determined not to be normally expressed. Certain diseases are known to be associated with CD38, such as tumors, especially cancer or leukemia.

[0074] Chronic lymphocytic leukemia (CLL): CLL is a common adult leukemia caused by the accumulation of small B lymphocytes (CD19+ / CD5+ / CD23+) in the blood, bone marrow, lymph nodes and other lymphoid tissues. The progression of the disease stage can be determined by detecting the increased expression of CD38.

[0075] Multiple myeloma (MM): MM is a malignant tumor characterized by the accumulation of monoclonal plasma cells, high concentrations of monoclonal immunoglobulins (Ig) in plasma, urine, and lytic bone marrow.

[0076] Acute promyelocytic leukemia (APL): APL is a distinct subtype of acute leukemia characterized by the suppression of leukocyte differentiation at its promyelocytic stage. Overexpression of CD38 on granulocytes can be observed in patients with the disease.

[0077] Other diseases: CD38 has been reported to be associated with non-Hodgkin's lymphoma, B- and T-cell acute lymphoblastic leukemia, acute myeloid leukemia, Hodgkin's lymphoma, and chronic myeloid leukemia.

[0078] Deletion, addition, substitution or insertion of amino acids: Amino acid substitution means replacement with known amino acids, preferably conservative amino acid substitution. Generally, substitution of conservative amino acid residues does not affect the mechanism of proteins or peptides. Examples of amino acid groups with side chains of similar chemical properties include: 1) aliphatic side chains: glycine, alanine, valine, leucine and isoleucine; 2) aliphatic hydroxyl side chains: serine and threonine; 3) amide-containing side chains: asparagine and glutamine; 4) aromatic side chains: phenylalanine, tyrosine and tryptophan; 5) basic side chains: lysine, arginine and histidine; 6) acidic side chains: aspartic acid and glutamic acid; 7) sulfur-containing side chains: including cysteine ​​and methionine. Conservative amino acid substitutions are: valine-leucine-isoleucine, phenylalanine-tyrosine-tryptophan, lysine-arginine, alanine-valine, glutamic acid-aspartic acid and asparagine-glutamine. The aforementioned amino acids may be proteinaceous or non-proteinaceous amino acids. Throughout the specification, the term "one or more amino acids" refers to one or more amino acids that may be deleted, substituted, inserted and / or added. In addition, in the present specification, the term "one or more amino acids" may refer to one or some amino acids in some cases. The present invention includes but is not limited to a peptide consisting of an amino acid sequence, which has 1 to 5, preferably 4 or less, 3 or less, 2 or less, more preferably one or less amino acids deleted, substituted, inserted and / or added in the amino acid sequence represented by one of SEQ ID NOs. 1 to 34, and has the activity of binding to CD38.

[0079] Surgery or treatment and diagnosis of CD38-related diseases: The present invention can treat CD38-related diseases by binding to CD38. For example, using a conjugate of the CD38-binding peptide of the present invention and a cytotoxic compound, it can directly or indirectly specifically kill or damage cells expressing CD38, especially cancer cells. At the same time, by detecting a marker conjugated to the CD38-binding peptide and administered to a patient, the distribution of CD38 expression can be detected in vivo. For example, the progression of the disease can be diagnosed by administering and detecting the amount of the CD38-binding peptide in a CLL patient.

[0080] Target: In some embodiments, the present invention provides techniques for selectively directing agents comprising a CD38 binding moiety to a target site comprising one or more desired targets. As will be appreciated by those skilled in the art, the provided techniques are useful for various types of targets, particularly targets comprising CD38.

[0081] In some embodiments, the target is damaged or defective tissue. In some embodiments, the target is damaged tissue. In some embodiments, the target is defective tissue. In some embodiments, the target is associated with a disease, disorder, or condition, such as cancer, wounds, etc. In some embodiments, the target is a tumor. In some embodiments, the target is a disease cell or comprises a disease cell. In some embodiments, the target is a cancer cell or comprises a cancer cell. In some embodiments, the target is a foreign object. In some embodiments, the target is an infectious agent or comprises an infectious agent. In some embodiments, the target is a microorganism. In some embodiments, the target is a bacterium or comprises a bacterium. In some embodiments, the target is a virus or comprises a virus. In some embodiments, the target comprises CD38 or expresses CD38.

[0082] In many embodiments, the target is a tissue and / or cell associated with a disease, disorder or condition, particularly tissue and / or cell associated with various types of cancer. In some embodiments, the target is a cell associated with a disease, disorder or condition or comprises a cell associated with a disease, disorder or condition. In some embodiments, the target is a cell associated with cancer or comprises a cell associated with cancer. In some embodiments, the cell comprises CD38 or expresses CD38.

[0083] Conjugate: In some embodiments, the present invention provides a reagent that is a conjugate. In some embodiments, the conjugate provided comprises: a first part that can bind to CD38; a second part that can be a variety of structures for various purposes such as a detection reagent, a diagnostic reagent or a therapeutic agent; and an optional linker that connects the first part and the second part.

[0084] In some embodiments, the CD38 binding peptide and the material desired to be delivered can be connected. The material desired to be delivered is not limited, for example, it can be a material used to treat CD38-related diseases. For example, it can be an anti-CD38 antibody, a cytotoxic compound, or a chemical material labeled with a radioactive isotope. It can also be a material that directly damages cells bound to the CD38 binding peptide, or a material that indirectly damages those cells, for example, an anticancer substance (such as daratumumab), which binds to the CD38 antigen expressed on the surface of cancer cells of hematopoietic tumors including multiple myeloma by binding to complement dependent cytotoxicity (CDC) or antibody dependent cellular cytotoxicity (ADCP), thereby affecting the cancer cells.

[0085] In addition, such materials can be any label used for diagnosing CD38-related diseases. For example, antibodies, bio-enhancing labels, PET reagents with guideable labels. For the sake of clarity, one or more amino acids of the infection can be directly or indirectly labeled when used for diagnosis.

[0086] In some embodiments, a CD38 binding moiety, such as the first moiety, is -PM or comprises -PM, wherein H-PM is a compound shown in Table 1 or a salt thereof. In some embodiments, H-PM is I-1 or a salt thereof. In some embodiments, H-PM is I-2 or a salt thereof. In some embodiments, a first moiety, such as -PM, is or comprises:

[0087] [Chemical formula 01]

[0088]

[0089] or a salt form thereof.

[0090] In some embodiments, a first portion, such as -PM, is or comprises

[0091] [Chemical formula 02]

[0092]

[0093] or a salt form thereof.

[0094] As will be appreciated by those skilled in the art, the provided conjugates may contain moieties of a variety of useful agents (e.g., as a second moiety) for a variety of purposes. Suitable agents / moieties for detection, diagnosis, treatment, etc. are well known in the art and may be used in accordance with the present invention.

[0095] For example, in some embodiments, useful agents are therapeutic agents. In some embodiments, useful agents are FDA approved therapeutic agents, or therapeutic agents that have been or are undergoing clinical trials. In some embodiments, useful agents are cancer therapeutic agents. In some embodiments, useful agents kill cells or inhibit cell growth. In some embodiments, useful agents are toxins, such as toxins that can inhibit cell growth and / or kill cells. In some embodiments, useful agents are radiopharmaceuticals.

[0096] In some embodiments, the second agent is selected from a small molecule, a nucleic acid, a polypeptide, a protein, a carbohydrate, a lipid, and any combination thereof. In some embodiments, the second part is a part of such a second agent.

[0097] In some embodiments, the second moiety is a small molecule payload of an antibody-drug conjugate, which has been extensively described in the art.

[0098] In some embodiments, the second portion is useful for detection and / or diagnosis, for example, a portion that is capable of providing a signal that can be detected using appropriate techniques under appropriate conditions. For example, in some embodiments, the second portion is a fluorescent portion or comprises a fluorescent portion. In some embodiments, the second portion is a radioactive label or comprises a radioactive label (e.g., an isotope for detection). In some embodiments, the second portion is a portion for imaging or comprises a portion for imaging, such as PET. In some embodiments, the second portion is an RI tag or comprises an RI tag. In some embodiments, the second portion is an RI imaging tag or comprises an RI imaging tag. In some embodiments, the second portion is an FDG tag or comprises an FDG tag. In some embodiments, the second portion is a paramagnetic ion tag or comprises a paramagnetic ion tag. In some embodiments, the second portion is a solid phase portion or comprises a solid phase portion. In some embodiments, the second portion is a metal nanotag or comprises a metal nanotag, such as an Au or Ag tag. In some embodiments, the second part is an affinity tag or comprises an affinity tag, for example, a His tag (poly-His, hexa-His) tag, an HA tag, a myc tag, a FLAG tag, a V5 tag, an S tag, an E tag, a T7 tag, a VSV-G tag, a Gly-Glu tag, an HSV tag, a Strep (II) tag, a CBD tag, a CBP tag, a GST (glutathione S-transferase) tag, an MBP (maltose binding protein) tag, a thioredoxin tag, a biotin carboxyl carrier protein (BCCP) tag, etc. In some embodiments, the second part is a metal fluorescent tag or comprises a metal fluorescent tag, such as a GFP tag, an RFP tag, a FITC tag, etc. In some embodiments, the second part is an incomplete antigen tag or comprises an incomplete antigen tag. In some embodiments, the second part is a label or comprises a label, such as a FLAG tag, an HA tag, etc. In some embodiments, the second part is a nucleic acid tag or comprises a nucleic acid tag.

[0099] Certain useful parts are described in WO2016 / 146639, WO2009 / 140408, WO2015 / 095895, WO2016 / 168817, WO2009 / 092732, Mol. BioSyst., 2016, 12, 1731-1745, Trends Biotechnol. 2012 January; 30(1): 8-16.

[0100] The conjugate can include one or more first moieties (eg, CD38 binding moieties) and one or more second moieties, and the first moieties can be the same or different, and the second moieties can be the same or different and can be used for the same or different purposes.

[0101] In some embodiments, the present invention provides a method for preventing or treating a state, disorder or disease associated with CD38, the method comprising administering an effective dose of a provided compound or a pharmaceutically acceptable salt thereof to a subject susceptible or affected therefrom. In some embodiments, the present invention provides a method for detecting a target comprising CD38 (e.g., a cell expressing CD38) in a system, the method comprising administering a provided compound or a salt thereof to the system. In some embodiments, the present invention provides a method for detecting a target comprising CD38 (e.g., a cell expressing CD38) in a system, the method comprising contacting the target with a provided compound or a salt thereof. In some embodiments, the present invention provides a method for diagnosing a state, disorder or disease, the method comprising administering a provided compound or a salt thereof to a subject. In some embodiments, the provided compound is a compound selected from the group consisting of compounds shown in Formulas I-1 to I-38. In some embodiments, the provided compound is a conjugate. In some embodiments, the provided compound is a conjugate of a compound shown in Formulas I-1 to I-38.

[0102] Various conjugated chemistries are well known and practiced in the art and can be used to prepare conjugates as provided by the present invention. For example, in some embodiments, the first portion is optionally connected to the first reactive group via a linker, and the second portion is optionally connected to the second reactive group via a linker, and the first reactive group can react with the second reactive group. In certain embodiments, the reactive group is or comprises an amino group, an acid group, a reactive group for cycloaddition reactions (e.g., Diels-Alder and variants thereof, Staudinger ligation reactions and variants thereof, keying chemistry and variants thereof, etc.) and an activated form thereof. In certain embodiments, the first reactive group is an amino group. In certain embodiments, the second reactive group is an activated acid.

[0103] Connection part

[0104] The CD38 binding peptide and materials to be delivered to CD38 can be linked to the linker.

[0105] The linker (also referred to as a cross-linking agent) may be any conventional linker or as described herein. In some embodiments, such linkers may be, for example, chemical linkers, lipid linkers, peptide (polypeptide) linkers. Alternatively, it may be a combination of chemical linkers and peptide linkers. For example, such linkers may be linkers that can dissociate or separate or stabilize under specific conditions. It may also be a PEG (polyethylene glycol) linker or a peptide linker. For example, it may be a PEG composed of 1-24 ethylene glycol units. The peptide linker may be composed of at least one amino acid. In some embodiments, the linker may be an amino acid (for example, it may be any amino acid, such as Gly, Lys, Glu, Ser or Ala). In some embodiments, the linker may be a combination of a PEG linker and a peptide linker, and the peptide linker is bound to a chemical compound.

[0106] Exemplary compounds include those listed in Table 1 below.

[0107] Table 1. Exemplary compounds [Chemical Formula 03]

[0108]

[0109] [Chemical formula 04]

[0110]

[0111] [Chemical formula 05]

[0112]

[0113] [Chemical formula 06]

[0114]

[0115] [Chemical formula 07]

[0116]

[0117] [Chemical formula 08]

[0118]

[0119] In some embodiments, the present invention provides compounds listed in Table 1 above or pharmaceutically acceptable salts thereof.

[0120] 4. General Methods for Providing Compounds of the Invention

[0121] In general, the compounds of the present invention can be prepared or isolated by synthetic and / or semi-synthetic methods known to those skilled in the art for similar compounds, as well as the methods described in detail in the Examples herein.

[0122] In some embodiments, where a specific protecting group ("PG"), leaving group ("LG"), or transformation condition is described, one of ordinary skill in the art will appreciate that other protecting groups, leaving groups, and transformation conditions are also suitable and can be considered. March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, MB Smith and J. March, 5 th Edition, John Wiley & Sons, 2001, Comprehensive Organic Transformations, RC Larock, 2 nd Edition, John Wiley & Sons, 1999, and Protecting Groups in Organic Synthesis, TW Greene and PGMWuts, 3 rd Such groups and transformations are described in detail in SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3. Such groups and transformations are described in detail in SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7. Edition, John Wiley & Sons, 1999, the entire contents of which are incorporated herein by reference.

[0123] In some embodiments, the leaving group includes but is not limited to halogen (e.g., fluoride, chloride, bromide, iodide), sulfonate (e.g., -S(O)2R, methanesulfonate, toluenesulfonate, benzenesulfonate, p-bromobenzenesulfonate, m-nitrobenzenesulfonate, trifluoromethanesulfonate), diazonium salt, and the like.

[0124] In some embodiments, the oxygen protecting group includes, for example, a carbonyl protecting group, a hydroxyl protecting group, etc. Hydroxyl protecting groups are well known in the art and are also included in Protecting Groups in Organic Synthesis, TW Greene and P.G.M. Wuts, 3 rdThe invention relates to hydroxyl protecting groups, for example, hydroxyl protecting groups, such as hydroxyl protecting groups, hydroxyl protecting groups, hydroxyl protecting groups, and hydroxyl protecting groups. The invention relates to hydroxyl protecting groups, for example, ... Specific examples include formates, benzoylformates, chloroacetates, trifluoroacetates, methoxyacetates, triphenylmethoxyacetates, p-chlorophenoxyacetates, 3-phenylpropionates, 4-oxopentanoates, 4,4-(ethylenedithio)pentanoates, pivaloates (pivaloyl), crotonates, 4-methoxycrotonates, benzoates, terephthalates, 2,4,6-trimethylbenzoate, carbonates such as methyl, 9-fluorenylmethyl, ethyl, 2,2,2-trichloroethyl, 2-(trimethylsilyl)ethyl, 2-(phenylsulfonyl)ethyl, vinyl, allyl, and p-nitrobenzyl. Examples of such silyl ethers include trimethylsilyl, triethylsilyl, tert-butyldimethylsilyl, tert-butyldiphenylsilyl, triisopropylsilyl, and other trialkylsilyl ethers. Alkyl ethers include methyl, benzyl, p-methoxybenzyl, 3,4-dimethoxybenzyl, trityl, tert-butyl, allyl, and allyloxycarbonyl ethers or derivatives. Alkoxyalkyl ethers include acetals such as methoxymethyl, methylthiomethyl, (2-methoxyethoxy)methyl, benzyloxymethyl, β-(trimethylsilyl)ethoxymethyl, and tetrahydropyranyl ether. Examples of aralkyl ethers include benzyl, p-methoxybenzyl (MPM), 3,4-dimethoxybenzyl, O-nitrobenzyl, p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl, and 2- and 4-pyridylmethyl.

[0125] Amino protecting groups are well known in the art and are included in Protecting Groups in Organic Synthesis, TW Greene and PGM Wuts, 3 rd The invention relates to amino protecting groups, for example, tert-butyloxycarbonyl (BOC), ethoxycarbonyl, methoxycarbonyl, trichloroethoxycarbonyl, allyloxycarbonyl (Alloc), benzyloxycarbonyl (CBZ), allyl, phthalimide, benzyl (Bn), fluorenylmethylcarbonyl (Fmoc), formyl, acetyl, chloroacetyl, dichloroacetyl, trichloroacetyl, phenylacetyl, trifluoroacetyl, benzoyl, etc.

[0126] Those skilled in the art will appreciate that compounds of Formula I or II may contain one or more stereocenters and may exist as racemic or diastereomeric mixtures. Those skilled in the art will also appreciate that many methods are known in the art for separating isomers to obtain stereoenriched or stereopure isomers of those compounds, including but not limited to HPLC, palm-to-palm HPLC, differential crystallization of diastereomeric salts, kinetic enzymatic hydrolysis (e.g., by fungal, bacterial or animal derived lipases or esterases), and the use of enantiomeric enrichment agents to form covalent diastereomeric derivatives.

[0127] Those skilled in the art will appreciate that the various functional groups present in the compounds of the present invention, such as aliphatic groups, alcohols, carboxylic acids, esters, amides, aldehydes, halogens and nitriles, can be interconverted by techniques well known in the art, including but not limited to reduction, oxidation, esterification, hydrolysis, partial oxidation, partial reduction, halogenation, dehydration, partial hydration and hydration. "March's Advanced Organic Chemistry", 5 th Ed., Ed.: Smith, MB and March, J., John Wiley & Sons, New York: 2001" is incorporated herein by reference in its entirety. Such interconversions can require one or more of the aforementioned techniques, and certain methods for synthesizing compounds of the invention are described below in the Examples.

[0128] Many compounds provided herein include peptide moieties. Such compounds can be prepared using a variety of techniques by systems comprising biological peptide synthesis mechanisms (e.g., in vivo or in vitro translation systems) or by chemical synthesis. In some embodiments, peptides are prepared by in vitro translation. In some embodiments, peptides are chemically synthesized, for example, by Fmoc chemistry. Useful techniques for Fmoc peptide synthesis, including amino acid reagents, protecting groups, coupling conditions, activating reagents, deprotection conditions, purification methods, etc., are well known in the art and can be easily used to prepare the compounds provided.

[0129] In some embodiments, the provided compounds include a cyclic peptide portion. According to the present invention, many suitable cyclization techniques can be utilized. For example, in some embodiments, an amino acid, such as an N-terminal amino acid, includes a leaving group (-LG, such as -Cl, -Br, -I, etc.), and can react with another amino acid, wherein the other amino acid includes a nucleophile (e.g., -SH of a cysteine ​​side chain). In some embodiments, as described herein, a leaving group is inserted into an amino acid via -C(O)CH2-LG, wherein the -C(O)CH2-LG is bonded to an amino group (e.g., an N-terminal amino acid residue). In some embodiments, -LG is -Cl. In some embodiments, after the peptide is released from protection, cyclization can be achieved under suitable conditions to provide a compound comprising a cyclic peptide portion. In some embodiments, the provided compound is of formula R CN -(Xaa)yR CS or a salt thereof, wherein R CN is as described herein, (Xaa)y is as described herein, R CS YesR CC or a scaffold, wherein each side chain of Xaa is independently and optionally protected. In some embodiments, R CN In some embodiments, R CN is -C(O)-LG. In some embodiments, R CN is -C(O)-Cl. In some embodiments, R CN Bonded to the N-terminal amino group. In some embodiments, R CS is a scaffold, such as a scaffold for peptide synthesis. In some embodiments, R CS is a resin suitable for peptide synthesis using Fmoc chemistry. In some embodiments, the C-terminal residue is a Cys residue, which is optionally protected.

[0130] As will be appreciated by those skilled in the art, a number of techniques can be used to prepare conjugates, including those utilizing the reactivity of amino groups, activated acid groups (e.g., NHS esters), nucleophiles, electrophiles, cycloaddition substrates, etc. In some embodiments, the reaction is a biocompatible reaction, which is well known and practiced in the art and can be used in accordance with the present invention.

[0131] The present invention particularly provides high purity compounds. In some embodiments, the provided compounds have a purity of 80-100%, for example at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%.

[0132] 5. Techniques for evaluating provided compounds

[0133] Various assays can be used to assess the properties and functions of the provided compounds. For example, various techniques, such as SPR, ELISA, display, FACS, etc. are useful for measuring the binding of the provided compounds to CD38. In some embodiments, the binding is assessed in a cell-based assay. Many methods including intracellular and extracellular can be used to measure the biological effects of the provided compounds and compositions. In some embodiments, for example, the Kd for the binding of the agent to CD38 is not greater than 1000, 500, 200, 150, 120, 100, 90, 80, 70, 60, 50, 40, 30, 20, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 nM, as measured by the assays described herein. In some embodiments, the Kd is not greater than 500 nM. In some embodiments, the Kd is not greater than 200 nM. In some embodiments, the Kd is not greater than 100 nM. In some embodiments, the Kd is not greater than 50 nM. In some embodiments, the Kd is not greater than 20 nM. In some embodiments, the Kd is not greater than 10 nM. In some embodiments, the Kd is no greater than 5 nM. In some embodiments, the Kd is no greater than 1 nM.

[0134] In some embodiments, provided compounds can be evaluated in comparison with many other compounds in a database, such as sequences containing many other peptides. In some embodiments, the database is a phage display database. In some embodiments, the database is an RNA, such as an mRNA display database. In some embodiments, the database is a DNA display database.

[0135] Useful related techniques include those described in US2016 / 0068835, US 9410148, US 8188260, and US 9090668. In at least one such test using a phage database, the peptide sequence of the provided compound exhibits higher binding affinity than other sequences.

[0136] 6. Use, formulation and administration

[0137] Pharmaceutically acceptable composition

[0138] According to another embodiment, the present invention provides a composition comprising a compound described herein or a pharmaceutically acceptable derivative thereof and a pharmaceutically acceptable carrier, adjuvant or excipient thereof. In some embodiments, the present invention provides a pharmaceutical composition comprising a compound of the present invention and a pharmaceutically acceptable carrier. In some embodiments, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of a compound and a pharmaceutically acceptable carrier.

[0139] In some embodiments, the pharmaceutically acceptable carrier, adjuvant or excipient is a non-toxic carrier, adjuvant or excipient that does not destroy the pharmacological activity of the compound formulated therewith. Pharmaceutically acceptable carriers, adjuvants or excipients may include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures containing saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silicon dioxide, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, waxes, polyethylene-polyoxypropylene block polymers, polyethylene glycol and lanolin.

[0140] In some embodiments, a pharmaceutically acceptable derivative is a non-toxic salt, ester, salt of an ester, or other derivative of the compound that, upon administration, is capable of providing, directly or indirectly, the compound or its active metabolite or residue.

[0141] The composition can be administered orally, parenterally, topically by inhalation spray through the rectum, nose, cheek, vagina, or via an implanted reservoir. In some embodiments, parenteral administration includes subcutaneous, intravenous, intramuscular, intraarticular, intrasynovial, intrasternal, intrathecal, intrahepatic, intralesional, and intracranial injection or infusion techniques. In some embodiments, the composition is administered orally, intraperitoneally, or intravenously. The sterile injection form of the composition can be an aqueous or oily suspension. Such suspensions can be prepared using suitable dispersants or wetting agents and suspending agents according to techniques known in the art. Sterile injection preparations can also be sterile injection solutions or suspensions in non-toxic parenteral acceptable diluents or solvents, such as solutions in 1,3-butanediol. Acceptable excipients and solvents can use water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile fixed oils are generally used as solvents or suspension media.

[0142] Uses of compounds and compositions thereof

[0143] The provided technology is useful for many purposes related to CD38. The provided compounds and compositions thereof are particularly useful for CD38 binding and modulating the properties and / or activities of CD38.

[0144] In some embodiments, the provided technology is particularly useful for delivering agents to targets comprising CD38 in vivo or in vitro for detection, diagnosis and / or treatment. In some embodiments, the agent is delivered via a conjugate comprising a CD38 binding moiety.

[0145] Provided compounds are particularly useful for preventing or treating various conditions, disorders or diseases associated with CD38. In some embodiments, the condition, disorder or disease is cancer.

[0146] In some embodiments, provided compounds are useful for labeling targets, such as cells, having CD38 for detection, isolation, characterization, or targeting by therapeutic agents (e.g., small molecule chemicals for PDC).

[0147] Example

[0148] As described in the following examples, in certain exemplary embodiments, compounds are prepared according to the following general procedures. It should be understood that although the general methods describe the synthesis of certain compounds of the present invention, the following general methods and other methods known to those of ordinary skill in the art can be applied to all compounds as described herein and their subclasses and species.

[0149] Example 1. Provided compounds bind CD38

[0150] A phage display database was constructed using techniques such as those described in US2016068835, US 9410148, and US 8188260 to evaluate the binding of many peptides to CD38. A peptide comprising the following sequence was identified:

[0151] Recombinant human CD38 protein was purchased from R&D Systems, Inc. In some embodiments, the following codons are used: ClAc-Ala: N-chloroacetyl-L-alanine; MePhe: N-methyl-L-phenylalanine; MeGly: N-methyl-glycine; Ahp: (S)-2-aminoheptanoic acid; Bph: (S)-3-([1,1'-biphenyl]-4-yl)-2-aminopropionic acid.

[0152] A number of compounds with similar structures to compound 1-1 (the difference is the amino acid sequence) were prepared and evaluated in an ELISA assay. Some data are shown below. The first sequence is 1-1 and SEQ ID. NO. 1.

[0153] [Table 2]

[0154]

[0155] *The larger the value, the stronger the bond.

[0156] A number of compounds with similar structures to compound 1-2 (the difference being the amino acid sequence) were prepared and evaluated in an ELISA assay. Some of the data are shown below. The first sequence is 1-2 and SEQ ID NO.2.

[0157] [Table 3]

[0158]

[0159] *The larger the value, the stronger the bond.

[0160] Example 2. Examples of binding constants

[0161] One of the many useful techniques for evaluating CD38 binding is surface plasmon resonance (SPR). For example, the binding constants of I-1 (SEQ ID NO. 1; same below) and I-2 (SEQ ID NO. 2; same below) were analyzed by SPR assay using BIACORE T200 (cytiva, formerly General Electric Healthcare). After equilibration of Series S Sensor Chip CM3 (cytiva, formerly General Electric Healthcare) with electrophoresis buffer (HBS-EP containing 1% (v / v) DMSO), the EDC / NHS mixture was injected at a flow rate of 10 μL / min for 4 minutes to activate the functional components on the sensor chip. CD38 dissolved in 10 mM acetate (pH 5.5) was fixedly injected at a flow rate of 5 μL / min. This was performed for 4 minutes to fix CD38 on the substrate surface of the sensor chip. Ethanolamine was then injected at a flow rate of 10 μL / min for 4 minutes. Then, 1.0 M ethanolamine (aqueous solution) was injected at a flow rate of 10 μL / min for 420 seconds for capping. After diluting the 10 mM peptide in DMSO with electrophoresis buffer to obtain 10 uM, peptide solutions (peptide samples) of various concentrations of 100 nM, 50 nM, 25 nM, 10 nM, and 5 nM were prepared.

[0162] The peptide samples were used to measure the kinetics of peptides against CD38. The method used for sample measurement was a single cycle kinetic method. The analysis was performed using the evaluation software attached to the Biacore T200. The DMSO calibration curve obtained by solvent correction measurement was provided for analysis. Kinetic fitting was performed for the difference data obtained by subtracting the baseline data from the sample measurement data.

[0163] The KD value was calculated based on the association rate constant (ka) and the dissociation rate constant (kd). The obtained results are shown in the table below.

[0164] [Table 4]

[0165] Chemical Ka Kd KD(M) I-1 (SEQ ID NO.1) <![CDATA[7.7*10 5 ]]> <![CDATA[1.8*10 -3 ]]> <![CDATA[2.3*10 -9 ]]> I-2 (SEQ ID NO.2) <![CDATA[3.6*10 5 ]]> <![CDATA[1.3*10 -2 ]]> <![CDATA[3.5*10 -8 ]]>

[0166] As described herein, provided compounds can effectively bind to CD38.

[0167] Example 3. Exemplary Synthesis of Provided Compounds

[0168] The abbreviations used herein are well known to those skilled in the art. Some abbreviations used are as follows: calcd. is calculation; Fmoc is fluorenylmethoxycarbonyl; HOAt is 1-hydroxy-7-azabenzotriazole; HATU is 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate; DMSO is dimethyl sulfoxide; DMF is N,N-dimethylformamide; mL is milliliter; M is mole; v / v is volume per volume; Mpe is methylpentan-3-yl (amino acid side chain protecting group); Pbf is (2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonyl (amino acid side chain protecting group); TFA is trifluoroacetic acid; TIS is triisopropylsilane; MeF is methylphenylalanine; Ahp is 2-aminoheptanoic acid; BPh is β-(4-biphenyl)-alanine. Some useful techniques are described in Bioconjugate Chem. 2018, 29, 1847-1851. In some embodiments, the peptide has C1Ac- at the N-terminus. In some embodiments, for cyclization, the deprotected peptide is dissolved in DMSO dissolved in 0.1% TFA, followed by addition of TEA until the solution is alkaline.

[0169] In general, peptides are synthesized by standard solid phase Fmoc chemistry. Peptides were assembled on Biotage Syro I (Biotage Japan Co., Ltd.). In the present invention, 9-Fmoc-amino-anthene-3-yloxo-Merrifield resin (Fmoc-protected Sieber resin) is used to prepare peptides with unsubstituted carboxamide at the C-terminus. The following lists some of the amino acid building blocks used in the cyclic peptide derivatives of the present invention, with side chain protecting groups shown in brackets. The listed amino acid building blocks are commercially available and can be used without further purification. Fmoc-Ahp-OH; Fmoc-Ala-OH; Fmoc-Arg(Pbf)-OH; Fmoc-Asp(OMpe)-OH; Fmoc-Cys(Trt)-OH; Fmoc-Gly-OH; Fmoc-His(Boc)-OH; Fmoc-L eu-OH; Fmoc-Lys(Boc)-OH; Fmoc-Pro-OH; Fmoc-Trp(Boc)-OH; Fmoc-Tyr(tBu)-OH; Fmoc-Val-OH; Fmoc-Arg-OH; Fmoc-Glu-OH; Fmoc-Gl y-OH; Fmoc-MetO2-OH; Fmoc-Hse-OH; Fmoc-Har-OH; Fmoc-D-Ala-OH; Fmoc-W6N-OH; Fmoc-Ano-OH; Fmoc-Ado-OH; Fmoc-PhNle-OH; Fmoc -PhNva-OH; Fmoc-Cit-OH; Fmoc-F3G-OH; Fmoc-FRNNMe-OH; Fmoc-RNNdMe-OH; Fmoc-RNdMe-OH; Fmoc-hCit-OH; Fmoc-4Py2NH2(Boc)-OH.

[0170] The peptide of the present invention was cycled by dissolving the peptide in DMSO to a final concentration of 5 mM, followed by the addition of 6 equivalents of TEA and stirring for about 16 hours. The reaction solution was neutralized with acetic acid and then vacuum concentrated with Biotage (trademark) V-10 (Biotage Japan Co., Ltd.).

[0171] Purification of the peptides of the present invention is performed by reverse phase HPLC, for example, using the conditions shown below: Stationary phase: Waters Xbridge TM , C18 5μm OBD TM , 50x250mm; mobile phase; washing liquid A (0.1% TFA dissolved in H2O), washing liquid B (0.1% TFA dissolved in CH3CN). The gradient is designed according to the specific requirements of the separation problem of each sample.

[0172] The structure of the peptides of the present invention is determined by, for example, ESI-MS (+). The purity of the peptides described herein is determined by the following analytical conditions, and the retention times are given in the described conditions for characterization purposes.

[0173] Analysis conditions

[0174] Stationary phase: Kinetex (trademark) EVO C18 100 Angstroms, 2.6 μm, 2.1 x 150 mm.

[0175] Mobile phase: Washing solution A (0.025% TFA in H2O); Washing solution B (0.025% TFA in CH3CN).

[0176] Column temperature: 60℃.

[0177] Gradient: See each experiment.

[0178] Detection method: UV 225nm.

[0179] Flow rate: 0.25mL / min.

[0180] abbreviation

[0181] [Table 5]

[0182]

[0183] Example 4. Exemplary Synthesis of I-1 (SEQ ID NO. 1)

[0184] [Chemical formula 09]

[0185]

[0186] The Fmoc amino acids used in the synthesis include Fmoc-Cys(Trt)-OH; Fmoc-Asp(OMpe)-OH; Fmoc-Ahp-OH; Fmoc-4-biphenyl-Ala-OH; Fmoc-Leu-OH; Fmoc-Val-OH; Fmoc-Gly-OH; Fmoc-His(Boc)-OH; Fmoc-Tyr(tBu)-OH; Fmoc-Arg(Pbf)-OH; Fmoc-Ala-OH. MS (ESI+); calcd. [M+2H] 2+ 832.9, found to be 833.3. Gradient for purity assessment: linear gradient, 20-60% wash solution B / wash solution A in 20 minutes. t ret = 8.7 minutes. Purity: 98.8%.

[0187] Example 5. Exemplary Synthesis of I-2 (SEQ ID NO. 2)

[0188] [Chemical formula 10]

[0189]

[0190] The Fmoc amino acids used in the synthesis include Fmoc-Cys(Trt)-OH; Fmoc-Val-OH; Fmoc-Trp(Boc)-OH; Fmoc-4-biphenyl-Ala-OH; Fmoc-Pro-OH; Fmoc-Leu-OH; Fmoc-N-Me-Phe-OH; Fmoc-His(Boc)-OH; Fmoc-Ala-OH. MS (ESI+); calcd. [M+2H] 2+ 780.4, found to be 780.8. Gradient for purity assessment: linear gradient, 20-60% wash solution B / wash solution A in 20 minutes. t ret = 8.6 minutes. Purity: 98.4%.

[0191] Example 6. Exemplary Synthesis of I-3

[0192] [Chemical formula 11]

[0193]

[0194] The Fmoc amino acids used in the synthesis include Fmoc-Lys(Boc)-OH; Fmoc-PEG-OH; Fmoc-Cys(Trt)-OH; Fmoc-Asp(OMpe)-OH; Fmoc-Ahp-OH; Fmoc-4-biphenyl-Ala-OH; Fmoc-Leu-OH; Fmoc-Val-OH; Fmoc-Gly-OH; Fmoc-His(Boc)-OH; Fmoc-Tyr(tBu)-OH; Fmoc-Arg(Pbf)-OH; Fmoc-Ala-OH. MS (ESI+); calcd. [M+2H] 2+ 1020.5, found to be 1021.0. Gradient for purity assessment: linear gradient, wash solution B / wash solution A 20-60% in 20 minutes. t ret = 7.9 minutes. Purity: 98.0%.

[0195] Example 7. Exemplary Synthesis of I-4

[0196] [Chemical formula 12]

[0197]

[0198] The Fmoc amino acids used in the synthesis include Fmoc-Lys(Boc)-OH; Fmoc-PEG-OH; Fmoc-Cys(Trt)-OH; Fmoc-Asp(OMpe)-OH; Fmoc-Ahp-OH; Fmoc-4-biphenyl-Ala-OH; Fmoc-Leu-OH; Fmoc-Val-OH; Fmoc-Gly-OH; Fmoc-His(Boc)-OH; Fmoc-Tyr(tBu)-OH; Fmoc-Arg(Pbf)-OH; Fmoc-Ala-OH. MS (ESI+); calcd. [M+2H] 2+ 1196.6, found to be 1197.2. Gradient for purity assessment: linear gradient, 20-60% wash solution B / wash solution A in 20 minutes. t ret = 8.6 minutes. Purity: 99.2%.

[0199] Example 8. Exemplary Synthesis of I-5

[0200] [Chemical formula 13]

[0201]

[0202] The Fmoc amino acids used in the synthesis include Fmoc-Lys(Boc(-OH; Fmoc-PEG-OH; Fmoc-Cys(Trt)-OH; Fmoc-Val-OH; Fmoc-Trp(Boc)-OH; Fmoc-4-phenylphenylalanine-OH; Fmoc-Pro-OH; Fmoc-Leu-OH; Fmoc-N-Me-Phe-OH; Fmoc-His(Boc)-OH; Fmoc-Ala-OH.MS(ESI+); calcd.[M+2H] 2+ 968.0, found to be 968.5. Gradient for purity assessment: linear gradient, 20-60% wash buffer B / wash buffer A in 20 minutes. t ret = 14.7 minutes. Purity: 98.0%.

[0203] Example 9. Exemplary Synthesis of I-6

[0204] [Chemical formula 14]

[0205]

[0206] The Fmoc amino acids used in the synthesis include Fmoc-Lys(Boc(-OH; Fmoc-PEG-OH; Fmoc-Cys(Trt)-OH; Fmoc-Val-OH; Fmoc-Trp(Boc)-OH; Fmoc-4-phenylphenylalanine-OH; Fmoc-Pro-OH; Fmoc-Leu-OH; Fmoc-N-Me-Phe-OH; Fmoc-His(Boc)-OH; Fmoc-Ala-OH.MS(ESI+); calcd.[M+2H] 2+ 1144.1, found to be 1144.7. Gradient for purity assessment; linear gradient: 20-60% washing solution B / washing solution A in 20 minutes. t ret = 15.3 minutes. Purity: 99.5%.

[0207] Example 10. Exemplary Synthesis of a Cyclic Peptide Having an Amino Acid Sequence Represented by SEQ ID NO. 3

[0208] [Chemical formula 15]

[0209]

[0210] The Fmoc amino acids used in the synthesis include Fmoc-Cys(Trt)-OH; Fmoc-Asp-OH; Fmoc-Ahp-OH; Fmoc-Bph-OH; Fmoc-Leu-OH; Fmoc-Val-OH; Fmoc-Ser-OH; Fmoc-His(Boc)-OH; Fmoc-Tyr(tBu)-OH; Fmoc-Arg(Pbf)-OH; and Fmoc-Ala-OH.

[0211] MS (ESI+); calcd. [M+2H] 2+ 847.5, found to be 848.4. Gradient for purity assessment: linear gradient, 20-60% wash buffer B / wash buffer A in 20 minutes. t ret = 3.98 minutes. Purity: 98.6%.

[0212] Example 11. Exemplary Synthesis of a Cyclic Peptide Having an Amino Acid Sequence Represented by SEQ ID NO.4

[0213] [Chemical formula 16]

[0214]

[0215] The Fmoc amino acids used in the synthesis include Fmoc-Cys(Trt)-OH; Fmoc-Asp-OH; Fmoc-Ahp-OH; Fmoc-Bph-OH; Fmoc-Leu-OH; Fmoc-Val-OH; Fmoc-Gln-OH; Fmoc-His(Boc)-OH; Fmoc-Tyr(tBu)-OH; Fmoc-Arg(Pbf)-OH; and Fmoc-Ala-OH.

[0216] MS (ESI+); calcd. [M+2H] 2+ 868.0, found to be 868.9. Gradient for purity assessment: linear gradient, 20-60% wash buffer B / wash buffer A in 20 minutes. t ret = 3.685 minutes. Purity: 95.5%.

[0217] Example 12. Exemplary Synthesis of a Cyclic Peptide Having an Amino Acid Sequence Represented by SEQ ID NO.5

[0218] [Chemical formula 17]

[0219]

[0220] The Fmoc amino acids used in the synthesis include Fmoc-Cys(Trt)-OH; Fmoc-Asp-OH; Fmoc-Ahp-OH; Fmoc-Bph-OH; Fmoc-Leu-OH; Fmoc-Val-OH; Fmoc-Asn-OH; Fmoc-His(Boc)-OH; Fmoc-Tyr(tBu)-OH; Fmoc-Arg(Pbf)-OH; and Fmoc-Ala-OH.

[0221] MS (ESI+); calcd. [M+2H] 2+ 868.0, found to be 861.9. Gradient for purity assessment: linear gradient, 20-60% wash buffer B / wash buffer A in 20 minutes. t ret = 3.75 minutes. Purity: 98.8%.

[0222] Example 13. Exemplary Synthesis of a Cyclic Peptide Having an Amino Acid Sequence Represented by SEQ ID NO.6

[0223] [Chemical formula 18]

[0224]

[0225] The Fmoc amino acids used in the synthesis include Fmoc-Cys(Trt)-OH; Fmoc-Asp-OH; Fmoc-Ahp-OH; Fmoc-Bph-OH; Fmoc-Leu-OH Fmoc-Val-OH; Fmoc-MetO2-OH; Fmoc-His(Boc)-OH; Fmoc-Tyr(tBu)-OH; Fmoc-Arg(Pbf)-OH; and Fmoc-Ala-OH.

[0226] MS (ESI+); calcd. [M+2H] 2+ 885.5, found to be 886.4. Gradient for purity assessment: linear gradient, 20-60% wash solution B / wash solution A in 20 minutes. t ret = 3.958 minutes. Purity: 97.9%.

[0227] Example 14. Exemplary Synthesis of a Cyclic Peptide Having an Amino Acid Sequence Represented by SEQ ID NO.7

[0228] [Chemical formula 19]

[0229]

[0230] The Fmoc amino acids used in the synthesis include Fmoc-Cys(Trt)-OH; Fmoc-Asp-OH; Fmoc-Ahp-OH; Fmoc-Bph-OH; Fmoc-Thr-OH; Fmoc-Val-OH; Fmoc-Gly-OH; Fmoc-His(Boc)-OH; Fmoc-Tyr(tBu)-OH; Fmoc-Arg(Pbf)-OH; and Fmoc-Ala-OH.

[0231] MS (ESI+); calcd. [M+2H] 2+ 826.4, found to be 827.4. Gradient for purity assessment: linear gradient, 20-60% wash solution B / wash solution A in 20 minutes. t ret = 3.25 minutes. Purity: 98.5%.

[0232] Example 15. Exemplary Synthesis of a Cyclic Peptide Having an Amino Acid Sequence Represented by SEQ ID NO.8

[0233] [Chemical formula 20]

[0234]

[0235] The Fmoc amino acids used in the synthesis include Fmoc-Cys(Trt)-OH; Fmoc-Asp-OH; Fmoc-Ahp-OH; Fmoc-Bph-OH; Fmoc-Hse-OH; Fmoc-Val-OH; Fmoc-Gly-OH; Fmoc-His(Boc)-OH; Fmoc-Tyr(tBu)-OH; Fmoc-Arg(Pbf)-OH; and Fmoc-Ala-OH.

[0236] MS (ESI+); calcd. [M+2H] 2+ 827.4, found to be 827.4. Gradient for purity assessment: linear gradient, wash solution B / wash solution A 20-60% in 20 minutes. t ret = 3.15 minutes. Purity: 98.9%.

[0237] Example 16. Exemplary Synthesis of a Cyclic Peptide Having an Amino Acid Sequence Represented by SEQ ID NO.9

[0238] [Chemical formula 21]

[0239]

[0240] The Fmoc amino acids used in the synthesis include Fmoc-Cys(Trt)-OH; Fmoc-Asp-OH; Fmoc-Ahp-OH; Fmoc-Bph-OH; Fmoc-MetO2-OH; Fmoc-Val-OH; Fmoc-Gly-OH; Fmoc-His(Boc)-OH; Fmoc-Tyr(tBu)-OH; Fmoc-Arg(Pbf)-OH; and Fmoc-Ala-OH.

[0241] MS (ESI+); calcd. [M+2H] 2+ 857.5, found 858.5. Gradient for purity assessment; linear gradient, 20-60% Wash B / Wash A in 20 minutes. t ret = 3.22 minutes. Purity: 99.1%.

[0242] Example 17. Exemplary Synthesis of a Cyclic Peptide Having an Amino Acid Sequence Represented by SEQ ID NO. 10

[0243] [Chemical formula 22]

[0244]

[0245] The Fmoc amino acids used in the synthesis include Fmoc-Cys(Trt)-OH; Fmoc-Asp-OH; Fmoc-Ahp-OH; Fmoc-Bph-OH; Fmoc-Leu-OH; Fmoc-Val-OH; Fmoc-Glu-OH; Fmoc-His(Boc)-OH; Fmoc-Tyr(tBu)-OH; Fmoc-Arg(Pbf)-OH; and Fmoc-Ala-OH.

[0246] MS (ESI+); calcd. [M+2H] 2+ 868.5, found to be 869.4. Gradient for purity assessment: linear gradient, 20-60% wash buffer B / wash buffer A in 20 minutes. t ret = 3.93 minutes. Purity: 95.9%.

[0247] Example 18. Exemplary Synthesis of a Cyclic Peptide Having an Amino Acid Sequence Represented by SEQ ID NO.11

[0248] [Chemical formula 23]

[0249]

[0250] The Fmoc amino acids used in the synthesis include Fmoc-Cys(Trt)-OH; Fmoc-Asp-OH; Fmoc-Ahp-OH; Fmoc-Bph-OH; Fmoc-Leu-OH; Fmoc-Val-OH; Fmoc-Glu-OH; Fmoc-His(Boc)-OH; Fmoc-Tyr(tBu)-OH; Fmoc-Arg(Pbf)-OH; and Fmoc-Ala-OH.

[0251] MS (ESI+); calcd. [M+2H] 2+ 882.0, found 883.0. Gradient for purity assessment: linear gradient, 20-60% wash buffer B / wash buffer A in 20 minutes. t ret = 3.29 minutes. Purity: 96.9%.

[0252] Example 19. Exemplary Synthesis of a Cyclic Peptide Having an Amino Acid Sequence Represented by SEQ ID NO. 12

[0253] [Chemical formula 24]

[0254]

[0255] The Fmoc amino acids used in the synthesis include Fmoc-Cys(Trt)-OH; Fmoc-Asp-OH; Fmoc-Ahp-OH; Fmoc-Bph-OH; Fmoc-Leu-OH; Fmoc-Val-OH; Fmoc-Har(Pbf)-OH; Fmoc-His(Boc)-OH; Fmoc-Tyr(tBu)-OH; Fmoc-Arg(Pbf)-OH; and Fmoc-Ala-OH.

[0256] MS (ESI+); calcd. [M+2H] 2+ 889.1, found to be 890.0. Gradient for purity assessment: linear gradient, 20-60% wash buffer B / wash buffer A in 20 minutes. t ret = 3.37 minutes. Purity: 96.5%.

[0257] Example 20. Exemplary Synthesis of a Cyclic Peptide Having an Amino Acid Sequence Represented by SEQ ID NO. 13

[0258] [Chemical formula 25]

[0259]

[0260] The Fmoc amino acids used in the synthesis include Fmoc-Cys(Trt)-OH; Fmoc-Asp-OH; Fmoc-Ahp-OH; Fmoc-Bph-OH; Fmoc-MetO2-OH; Fmoc-Val-OH; Fmoc-Gln-OH; Fmoc-His(Boc)-OH; Fmoc-Tyr(tBu)-OH; Fmoc-Arg(Pbf)-OH; and Fmoc-Ala-OH.

[0261] MS (ESI+); calcd. [M+2H] 2+ 893.1, found to be 894.0. Gradient for purity assessment: linear gradient, 20-60% wash buffer B / wash buffer A in 20 minutes. t ret = 3.62 minutes. Purity: 95.4%

[0262] Example 21. Exemplary Synthesis of a Cyclic Peptide Having an Amino Acid Sequence Represented by SEQ ID NO. 14

[0263] [Chemical formula 26]

[0264]

[0265] The Fmoc amino acids used in the synthesis include Fmoc-Cys(Trt)-OH; Fmoc-Asp-OH; Fmoc-Ahp-OH; Fmoc-Bph-OH; Fmoc-MetO2-OH; Fmoc-Val-OH; Fmoc-Glu-OH; Fmoc-His(Boc)-OH; Fmoc-Tyr(tBu)-OH; Fmoc-Arg(Pbf)-OH; and Fmoc-Ala-OH.

[0266] MS (ESI+); calcd. [M+2H] 2+ 893.5, found to be 894.4. Gradient for purity assessment: linear gradient, 20-60% wash buffer B / wash buffer A in 20 minutes. t ret = 3.69 minutes. Purity: 98.0%.

[0267] Example 22. Exemplary Synthesis of a Cyclic Peptide Having an Amino Acid Sequence Represented by SEQ ID NO. 15

[0268] [Chemical formula 27]

[0269]

[0270] The Fmoc amino acids used in the synthesis include Fmoc-Cys(Trt)-OH; Fmoc-Asp-OH; Fmoc-Ahp-OH; Fmoc-Bph-OH; Fmoc-MetO2-OH; Fmoc-Val-OH; Fmoc-Arg-OH; Fmoc-His(Boc)-OH; Fmoc-Tyr(tBu)-OH; Fmoc-Arg(Pbf)-OH; and Fmoc-Ala-OH.

[0271] MS (ESI+); calcd. [M+2H] 2+ 907.06, found 908.0. Gradient for purity assessment: linear gradient, 20-60% wash buffer B / wash buffer A in 20 minutes. t ret = 5.8 minutes. Purity: 96.8%.

[0272] Example 23. Exemplary Synthesis of a Cyclic Peptide Having an Amino Acid Sequence Represented by SEQ ID NO. 16

[0273] [Chemical formula 28]

[0274]

[0275] The Fmoc amino acids used in the synthesis include Fmoc-Cys(Trt)-OH; Fmoc-Asp-OH; Fmoc-Ahp-OH; Fmoc-Bph-OH; Fmoc-MetO2-OH; Fmoc-Val-OH; Fmoc-Har(Pbf)-OH; Fmoc-His(Boc)-OH; Fmoc-Tyr(tBu)-OH; Fmoc-Arg(Pbf)-OH; and Fmoc-Ala-OH.

[0276] MS (ESI+); calcd. [M+2H] 2+ 914.1, found to be 915.0. Gradient for purity assessment: linear gradient, 20-60% wash buffer B / wash buffer A in 20 minutes. t ret = 5.88 minutes. Purity: 96.8%.

[0277] Example 24. Exemplary Synthesis of a Cyclic Peptide Having an Amino Acid Sequence Represented by SEQ ID NO. 17

[0278] [Chemical formula 29]

[0279]

[0280] The Fmoc amino acids used in the synthesis include Fmoc-Cys(Trt)-OH; Fmoc-Asp-OH; Fmoc-Ahp-OH; Fmoc-Bph-OH; Fmoc-Leu-OH; Fmoc-Val-OH; Fmoc-Glu-OH; Fmoc-His(Boc)-OH; Fmoc-Tyr(tBu)-OH; Fmoc-Arg(Pbf)-OH; Fmoc-Lys-OH. Then, the side chain of the last Lys was modified by PEGylation using PEG (CAS No. 1188295-19-3).

[0281] MS (ESI+); calcd. [M+2H] 2+ 1006.2, found to be 1006.7. Gradient for purity assessment: linear gradient, wash solution B / wash solution A 20-60% in 20 minutes. t ret = 3.94 minutes. Purity: 91.6%.

[0282] Example 25. Exemplary Synthesis of a Cyclic Peptide Having an Amino Acid Sequence Represented by SEQ ID NO. 18

[0283] [Chemical formula 30]

[0284]

[0285] The Fmoc amino acids used in the synthesis include Fmoc-Cys(Trt)-OH; Fmoc-Asp-OH; Fmoc-Ahp-OH; Fmoc-Bph-OH; Fmoc-Leu-OH; Fmoc-Val-OH; Fmoc-Glu-OH; Fmoc-His(Boc)-OH; Fmoc-Tyr(tBu)-OH; Fmoc-Arg(Pbf)-OH; Fmoc-Lys-OH, Fmoc-Ala-OH. Next, the side chain of Lys at the second position was modified by PEGylation using PEG (CAS No. 1188295-19-3).

[0286] MS (ESI+); calcd. [M+2H] 2+ 963.6, found to be 964.6. Gradient for purity assessment: linear gradient, 20-60% wash solution B / wash solution A in 20 minutes. t ret = 4.71 minutes. Purity: 92.9%.

[0287] Example 26. Exemplary Synthesis of a Cyclic Peptide Having an Amino Acid Sequence Represented by SEQ ID NO. 19

[0288] [Chemical formula 31]

[0289]

[0290] The Fmoc amino acids used in the synthesis include Fmoc-Cys(Trt)-OH; Fmoc-Asp-OH; Fmoc-Ahp-OH; Fmoc-Bph-OH; Fmoc-Leu-OH; Fmoc-Val-OH; Fmoc-Glu-OH; Fmoc-His(Boc)-OH; Fmoc-Tyr(tBu)-OH; Fmoc-Arg(Pbf)-OH; Fmoc-Lys-OH, Fmoc-Ala-OH. Next, the side chain of Lys at the third position was modified by PEGylation using PEG (CAS No. 1188295-19-3).

[0291] MS (ESI+); calcd. [M+2H] 2+ 978.1, found to be 979.1. Gradient for purity assessment: linear gradient, 20-60% wash buffer B / wash buffer A in 20 minutes. t ret = 3.75 minutes. Purity: 86.9%.

[0292] Example 27. Exemplary Synthesis of a Cyclic Peptide Having an Amino Acid Sequence Represented by SEQ ID NO. 20

[0293] [Chemical formula 32]

[0294]

[0295] The Fmoc amino acids used in the synthesis include Fmoc-Cys(Trt)-OH; Fmoc-Asp-OH; Fmoc-Ahp-OH; Fmoc-Bph-OH; Fmoc-Leu-OH; Fmoc-Val-OH; Fmoc-Glu-OH; Fmoc-His(Boc)-OH; Fmoc-Tyr(tBu)-OH; Fmoc-Arg(Pbf)-OH, and Fmoc-D-Ala-OH.

[0296] MS (ESI+); calcd. [M+2H] 2+ 868.5, found to be 869.2. Gradient for purity assessment: linear gradient, 20-60% wash solution B / wash solution A in 20 minutes. t ret = 4.10 min. Purity: 97.1%.

[0297] Example 28. Exemplary Synthesis of a Cyclic Peptide Having an Amino Acid Sequence Represented by SEQ ID NO. 21

[0298] [Chemical formula 33]

[0299]

[0300] The Fmoc amino acids used in the synthesis include Fmoc-Cys(Trt)-OH; Fmoc-Asp-OH; Fmoc-Ahp-OH; Fmoc-Bph-OH; Fmoc-Leu-OH; Fmoc-Val-OH; Fmoc-Glu-OH; Fmoc-His(Boc)-OH; Fmoc-Ala-OH; and Fmoc-Arg(Pbf)-OH.

[0301] MS (ESI+); calcd. [M+2H] 2+ 822.5, found to be 823.2. Gradient for purity assessment: linear gradient, 20-60% wash solution B / wash solution A in 20 minutes. t ret = 3.96 minutes. Purity: 95.0%.

[0302] Example 29. Exemplary Synthesis of a Cyclic Peptide Having an Amino Acid Sequence Represented by SEQ ID NO. 22

[0303] [Chemical formula 34]

[0304]

[0305] The moc amino acids used in the synthesis include Fmoc-Cys(Trt)-OH; Fmoc-Asp-OH; Fmoc-Ahp-OH; Fmoc-Bph-OH; Fmoc-Leu-OH; Fmoc-Val-OH; Fmoc-Glu-OH; Fmoc-W6N-OH; Fmoc-Ala-OH; and Fmoc-Arg(Pbf)-OH.

[0306] MS (ESI+); calcd. [M+2H] 2+ 893.5, found to be 894.4. Gradient for purity assessment: linear gradient, 20-60% wash buffer B / wash buffer A in 20 minutes. t ret = 3.67 minutes. Purity: 97.3%.

[0307] Example 30. Exemplary Synthesis of a Cyclic Peptide Having an Amino Acid Sequence Represented by SEQ ID NO. 23

[0308] [Chemical formula 35]

[0309]

[0310] The Fmoc amino acids used in the synthesis include Fmoc-Cys(Trt)-OH; Fmoc-Ahp-OH; Fmoc-Asp-OH; Fmoc-Ano-OH; Fmoc-Bph-OH; Fmoc-Leu-OH; Fmoc-Val-OH; Fmoc-Glu-OH; Fmoc-His(Boc)-OH; Fmoc-Tyr(tBu)-OH; Fmoc-Ala-OH; and Fmoc-Arg(Pbf)-OH.

[0311] MS (ESI+); calcd. [M+2H] 2+ 882.5, found to be 883.2. Gradient for purity assessment: linear gradient, 20-60% wash solution B / wash solution A in 20 minutes. t ret = 4.38 minutes. Purity: 94.7%.

[0312] Example 31. Exemplary Synthesis of a Cyclic Peptide Having an Amino Acid Sequence Represented by SEQ ID NO. 24

[0313] [Chemical formula 36]

[0314]

[0315] The Fmoc amino acids used in the synthesis include Fmoc-Cys(Trt)-OH; Fmoc-Ahp-OH; Fmoc-Asp-OH; Fmoc-Ado-OH; Fmoc-Bph-OH; Fmoc-Leu-OH; Fmoc-Val-OH; Fmoc-Glu-OH; Fmoc-His(Boc)-OH; Fmoc-Tyr(tBu)-OH; Fmoc-Ala-OH; and Fmoc-Arg(Pbf)-OH.

[0316] MS (ESI+); calcd. [M+2H] 2+ 896.6, found to be 897.5. Gradient for purity assessment: linear gradient, 20-60% wash buffer B / wash buffer A in 20 minutes. t ret = 4.97 minutes. Purity: 88.7%.

[0317] Example 32. Exemplary Synthesis of a Cyclic Peptide Having an Amino Acid Sequence Represented by SEQ ID NO. 25

[0318] [Chemical formula 37]

[0319]

[0320] The Fmoc amino acids used in the synthesis include Fmoc-Cys(Trt)-OH; Fmoc-Ahp-OH; Fmoc-Asp-OH; Fmoc-PhNle-OH; Fmoc-Bph-OH; Fmoc-Leu-OH; Fmoc-Val-OH; Fmoc-Glu-OH; Fmoc-His(Boc)-OH; Fmoc-Tyr(tBu)-OH; Fmoc-Ala-OH; and Fmoc-Arg(Pbf)-OH.

[0321] MS (ESI+); calcd. [M+2H] 2+ 899.5, found to be 900.2. Gradient for purity assessment: linear gradient, 20-60% wash solution B / wash solution A in 20 minutes. t ret = 4.28 minutes. Purity: 97.1%.

[0322] Example 33. Exemplary Synthesis of a Cyclic Peptide Having an Amino Acid Sequence Represented by SEQ ID NO. 26

[0323] [Chemical formula 38]

[0324]

[0325] The Fmoc amino acids used in the synthesis include Fmoc-Cys(Trt)-OH; Fmoc-Ahp-OH; Fmoc-Asp-OH; Fmoc-PhNva-OH; Fmoc-Bph-OH; Fmoc-Leu-OH; Fmoc-Val-OH; Fmoc-Glu-OH; Fmoc-His(Boc)-OH; Fmoc-Tyr(tBu)-OH; Fmoc-Ala-OH; and Fmoc-Arg(Pbf)-OH.

[0326] MS (ESI+); calcd. [M+2H] 2+ 892.5, found to be 893.2. Gradient for purity assessment: linear gradient, 20-60% wash solution B / wash solution A in 20 minutes. t ret = 4.05 minutes. Purity: 95.8%.

[0327] Example 34. Exemplary Synthesis of SEQ ID NO. 27

[0328] [Chemical formula 39]

[0329]

[0330] The Fmoc amino acids used in the synthesis include Fmoc-Cys(Trt)-OH; Fmoc-Ahp-OH; Fmoc-Asp-OH; Fmoc-Bph-OH; Fmoc-Leu-OH; Fmoc-Val-OH; Fmoc-Glu-OH; Fmoc-His(Boc)-OH; Fmoc-Tyr(tBu)-OH; Fmoc-Ala-OH; and Fmoc-Arg(Pbf)-OH.

[0331] MS (ESI+); calcd. [M+2H] 2+ 846.5, found to be 847.4. Gradient for purity assessment: linear gradient, 20-60% wash buffer B / wash buffer A in 20 minutes. t ret = 4.05 minutes. Purity: 91.0%.

[0332] Example 35. Exemplary Synthesis of a Cyclic Peptide Having an Amino Acid Sequence Represented by SEQ ID NO. 28

[0333] [Chemical formula 40]

[0334]

[0335] The Fmoc amino acids used in the synthesis include Fmoc-Cit-OH; Fmoc-Ahp-OH; Fmoc-Cys(Trt)-OH; Fmoc-Asp-OH; Fmoc-Bph-OH; Fmoc-Leu-OH; Fmoc-Val-OH; Fmoc-Glu-OH; Fmoc-His(Boc)-OH; Fmoc-Tyr(tBu)-OH; and Fmoc-Ala-OH.

[0336] MS (ESI+); calcd. [M+2H] 2+ 900.04, found to be 900.9. Gradient for purity assessment: linear gradient, wash solution B / wash solution A 20-60% in 20 minutes. t ret = 4.85 minutes. Purity: 93.6%.

[0337] Example 36. Exemplary Synthesis of a Cyclic Peptide Having an Amino Acid Sequence Represented by SEQ ID NO. 29

[0338] [Chemical formula 41]

[0339]

[0340] The Fmoc amino acids used in the synthesis include Fmoc-F3G-OH; Fmoc-Ahp-OH; Fmoc-Cys(Trt)-OH; Fmoc-Asp-OH; Fmoc-Bph-OH; Fmoc-Leu-OH; Fmoc-Val-OH; Fmoc-Glu-OH; Fmoc-His(Boc)-OH; Fmoc-Tyr(tBu)-OH; and Fmoc-Ala-OH.

[0341] MS (ESI+); calcd. [M+2H] 2+ 923.6, found to be 924.3. Gradient for purity assessment: linear gradient, 20-60% wash buffer B / wash buffer A in 20 minutes. t ret = 3.95 minutes. Purity: 97.1%.

[0342] Example 37. Exemplary Synthesis of a Cyclic Peptide Having an Amino Acid Sequence Represented by SEQ ID NO. 30

[0343] [Chemical formula 42]

[0344]

[0345] The Fmoc amino acids used in the synthesis include Fmoc-FRNNMe-OH; Fmoc-Ahp-OH; Fmoc-Cys(Trt)-OH; Fmoc-Asp-OH; Fmoc-Bph-OH; Fmoc-Leu-OH; Fmoc-Val-OH; Fmoc-Glu-OH; Fmoc-His(Boc)-OH; Fmoc-Tyr(tBu)-OH; and Fmoc-Ala-OH.

[0346] MS (ESI+); calcd. [M+2H] 2+ 906.6, found to be 907.3. Gradient for purity assessment: linear gradient, 20-60% wash solution B / wash solution A in 20 minutes. t ret = 4.16 minutes. Purity: 94.1%.

[0347] Example 38. Exemplary Synthesis of a Cyclic Peptide Having an Amino Acid Sequence Represented by SEQ ID NO. 31

[0348] [Chemical formula 43]

[0349]

[0350] The Fmoc amino acids used in the synthesis include Fmoc-RNNdMe-OH; Fmoc-Ahp-OH; Fmoc-Cys(Trt)-OH; Fmoc-Asp-OH; Fmoc-Bph-OH; Fmoc-Leu-OH; Fmoc-Val-OH; Fmoc-Glu-OH; Fmoc-His(Boc)-OH; Fmoc-Tyr(tBu)-OH; and Fmoc-Ala-OH.

[0351] MS (ESI+); calcd. [M+2H] 2+ 913.6, found to be 914.63. Gradient for purity assessment: linear gradient, 20-60% wash solution B / wash solution A in 20 minutes. t ret = 3.33 minutes. Purity: 94.9%.

[0352] Example 39. Exemplary Synthesis of a Cyclic Peptide Having an Amino Acid Sequence Represented by SEQ ID NO. 32

[0353] [Chemical formula 44]

[0354]

[0355] The Fmoc amino acids used in the synthesis include Fmoc-RNdMe-OH; Fmoc-Ahp-OH; Fmoc-Cys(Trt)-OH; Fmoc-Asp-OH; Fmoc-Bph-OH; Fmoc-Leu-OH; Fmoc-Val-OH; Fmoc-Glu-OH; Fmoc-His(Boc)-OH; Fmoc-Tyr(tBu)-OH; and Fmoc-Ala-OH.

[0356] MS (ESI+); calcd. [M+2H] 2+ 913.6, found to be 914.2. Gradient for purity assessment: linear gradient, 20-60% wash solution B / wash solution A in 20 minutes. t ret = 4.40 min. Purity: 96.9%.

[0357] Example 40. Exemplary Synthesis of a Cyclic Peptide Having an Amino Acid Sequence Represented by SEQ ID NO. 33

[0358] [Chemical formula 45]

[0359]

[0360] The Fmoc amino acids used in the synthesis include Fmoc-hCit-OH; Fmoc-Ahp-OH; Fmoc-Cys(Trt)-OH; Fmoc-Asp-OH; Fmoc-Bph-OH; Fmoc-Leu-OH; Fmoc-Val-OH; Fmoc-Glu-OH; Fmoc-His(Boc)-OH; Fmoc-Tyr(tBu)-OH; and Fmoc-Ala-OH.

[0361] MS (ESI+); calcd. [M+2H] 2+ 907.4, found to be 907.9. Gradient for purity assessment: linear gradient, 20-60% wash buffer B / wash buffer A in 20 minutes. t ret = 5.01 min. Purity: 96.9%.

[0362] Example 41. Exemplary Synthesis of a Cyclic Peptide Having an Amino Acid Sequence Represented by SEQ ID NO. 34

[0363] [Chemical formula 46]

[0364]

[0365] The Fmoc amino acids used in the synthesis include Fmoc-4Py2NH2(Boc)-OH; Fmoc-Ahp-OH; Fmoc-Cys(Trt)-OH; Fmoc-Asp-OH; Fmoc-Bph-OH; Fmoc-Leu-OH; Fmoc-Val-OH; Fmoc-Glu-OH; Fmoc-His(Boc)-OH; Fmoc-Tyr(tBu)-OH; and Fmoc-Ala-OH.

[0366] MS (ESI+); calcd. [M+2H] 2+ 903.0, found to be 903.9. Gradient for purity assessment: linear gradient, 20-60% wash solution B / wash solution A in 20 minutes. t ret = 3.95 minutes. Purity: 98.4%.

[0367] Example 42. Binding Constants of PEGylated Compounds

[0368] The binding constants of the PEGylated compounds were analyzed by surface plasmon resonance as described in Example 2. The KDs of I-3, I-4, I-5, and I-6 were 3.5*10 -9 ,4.3*10 -9 , 10.5*10 -9 , 11.7*10 -9 M. As demonstrated, provided compounds comprising a CD38 binding peptide bound to a linker can bind strongly to CD38.

[0369] SPR interaction analysis using Biacore (Cytiva, formerly GE Healthcare)

[0370] The interaction analysis of SEQ ID NO.1 variants was performed as follows

[0371] SPR analysis was performed using Biacore S200 (Cytiva, formerly GE Healthcare)

[0372] After equilibration of the S series sensor chip NTA (manufactured by Cytiva, formerly General Electric Healthcare) with electrophoresis buffer (10 mM HEPES pH 7.4, 150 mM NaCl, 0.05% Tween 20), 350 mM EDTA was injected at a flow rate of 10 μL / min for 60 seconds, and 0.5 uM NiCl2 was injected at a flow rate of 10 μL / min for 60 seconds, and then 3 mM EDTA was injected at a flow rate of 10 μL / min for 120 seconds, and then the sensor chip was cleaned.

[0373] The EDC / NHS mixture was injected at a flow rate of 10 μL / min for 7 minutes to activate the functional groups on the sensor chip. His-tagged CD38 in electrophoresis buffer (10 mM HEPES pH 7.4, 150 mM NaCl, 0.05% Tween 20) was injected at a flow rate of 10 μL / min. CD38 was fixed on the substrate surface of the sensor chip for 7 minutes. Ethanolamine was then injected at a flow rate of 10 μL / min for 7 minutes.

[0374] 1.0 M ethanolamine (aqueous solution) was injected at a flow rate of 10 μL / min for 420 seconds for capping. 10 mM peptide (synthesized by the above method) in DMSO was diluted to 10 uM with electrophoresis buffer, and then 100 nM, 50 nM, 25 nM, 10 nM, and 5 nM peptide solutions (peptide samples) were prepared.

[0375] Using these peptide samples, the kinetics of peptides against His-tagged CD38 were determined. The method used for sample measurement was a single cycle kinetic method. The analysis was performed using the evaluation software attached to the Biacore S200. The DMSO calibration curve obtained by solvent correction measurement was provided for analysis. Kinetic fitting was performed for the difference data obtained by subtracting the baseline data from the sample measurement data.

[0376] The KD value was calculated based on the association rate constant (ka) and the dissociation rate constant (kd). The results are shown in the table below. As described herein, the provided compounds can effectively bind to CD38.

[0377] [Table 6]

[0378]

[0379] [Table 7]

[0380]

[0381] Industrial Applicability

[0382] Although we have described several embodiments, it is apparent that our basic embodiments can be modified to provide other embodiments utilizing the compounds and methods of the invention. It will be understood, therefore, that the scope of the invention is defined by the claims rather than by the specific examples shown in the examples. [Sequence Listing] <110> Peptide Dream Co., Ltd. <120> CD38 binding agents and uses thereof <130> 109E0254 <140> 109121703 <141> 2020-06-24 <150> US62 / 870629 <151> 2019-07-03 <160> 34 <170> PatentIn version 3.5 <210> 1 <211> 13 <212> PRT <213> Synthetic Peptides <220> <221> MISC_FEATURE <222> (3) <223> Xaa is Ahp <220> <221> MISC_FEATURE <222> (10)..(10) <223> Xaa is Bph <220> <221> MISC_FEATURE <222> (11)..(11) <223> Xaa is Ahp <400> 1 Ala Arg Xaa Tyr His Asp Gly Val Leu Xaa Xaa Asp Cys 1 5 10 <210> 2 <211> 12 <212> PRT <213> Synthetic Peptides <220> <221> MISC_FEATURE <222> (4) <223> Xaa is MePhe <220> <221> MISC_FEATURE <222> (8) <223> Xaa is Bph <220> <221> MISC_FEATURE <222> (11)..(11) <223> Xaa is Ahp <400> 2 Ala Leu His Xaa Val Leu Pro Xaa Val Trp Val Cys 1 5 10 <210> 3 <211> 13 <212> PRT <213> Synthetic Peptides <220> <221> MISC_FEATURE <222> (3) <223> Xaa is Ahp <220> <221> MISC_FEATURE <222> (10)..(10) <223> Xaa is Bph <220> <221> MISC_FEATURE <222> (11)..(11) <223> Xaa is Ahp <400> 3 Ala Arg Xaa Tyr His Asp Ser Val Leu Xaa Xaa Asp Cys 1 5 10 <210> 4 <211> 13 <212> PRT <213> Synthetic Peptides <220> <221> MISC_FEATURE <222> (3) <223> Xaa is Ahp <220> <221> MISC_FEATURE <222> (10)..(10) <223> Xaa is Bph <220> <221> MISC_FEATURE <222> (11)..(11) <223> Xaa is Ahp <400> 4 Ala Arg Xaa Tyr His Asp Gln Val Leu Xaa Xaa Asp Cys 1 5 10 <210> 5 <211> 13 <212> PRT <213> Synthetic Peptides <220> <221> MISC_FEATURE <222> (3) <223> Xaa is Ahp <220> <221> MISC_FEATURE <222> (10)..(10) <223> Xaa is Bph <220> <221> MISC_FEATURE <222> (11)..(11) <223> Xaa is Ahp <400> 5 Ala Arg Xaa Tyr His Asp Asn Val Leu Xaa Xaa Asp Cys 1 5 10 <210> 6 <211> 13 <212> PRT <213> Synthetic Peptides <220> <221> MISC_FEATURE <222> (3) <223> Xaa is Ahp <220> <221> MISC_FEATURE <222> (7) <223> Xaa is MetO2 <220> <221> MISC_FEATURE <222> (10)..(10) <223> Xaa is Bph <220> <221> MISC_FEATURE <222> (11)..(11) <223> Xaa is Ahp <400> 6 Ala Arg Xaa Tyr His Asp Xaa Val Leu Xaa Xaa Asp Cys 1 5 10 <210> 7 <211> 13 <212> PRT <213> Synthetic Peptides <220> <221> MISC_FEATURE <222> (3) <223> Xaa is Ahp <220> <221> MISC_FEATURE <222> (10)..(10) <223> Xaa is Bph <220> <221> MISC_FEATURE <222> (11)..(11) <223> Xaa is Ahp <400> 7 Ala Arg Xaa Tyr His Asp Gly Val Thr Xaa Xaa Asp Cys 1 5 10 <210> 8 <211> 13 <212> PRT <213> Synthetic Peptides <220> <221> MISC_FEATURE <222> (3) <223> Xaa is Ahp <220> <221> MISC_FEATURE <222> (9)..(9) <223> Xaa is Hse <220> <221> MISC_FEATURE <222> (10)..(10) <223> Xaa is Bph <220> <221> MISC_FEATURE <222> (11)..(11) <223> Xaa is Ahp <400> 8 Ala Arg Xaa Tyr His Asp Gly Val Xaa Xaa Xaa Asp Cys 1 5 10 <210> 9 <211> 13 <212> PRT <213> Synthetic peptide <220> <221> MISC_FEATURE <222> (3)..(3) <223> Xaa is Ahp <220> <221> MISC_FEATURE <222> (9)..(9) <223> Xaa is MetO2 <220> <221> MISC_FEATURE <222> (10)..(10) <223> Xaa is Bph <220> <221> MISC_FEATURE <222> (11)..(11) <223> Xaa is Ahp <400> 9 Ala Arg Xaa Tyr His Asp Gly Val Xaa Xaa Xaa Asp Cys 1 5 10 <210> 10 <211> 13 <212> PRT <213> Synthetic Peptides <220> <221> MISC_FEATURE <222> (3) <223> Xaa is Ahp <220> <221> MISC_FEATURE <222> (10)..(10) <223> Xaa is Bph <220> <221> MISC_FEATURE <222> (11)..(11) <223> Xaa is Ahp <400> 10 Ala Arg Xaa Tyr His Asp Glu Val Leu Xaa Xaa Asp Cys 1 5 10 <210> 11 <211> 13 <212> PRT <213> Synthetic Peptides <220> <221> MISC_FEATURE <222> (3) <223> Xaa is Ahp <220> <221> MISC_FEATURE <222> (10)..(10) <223> Xaa is Bph <220> <221> MISC_FEATURE <222> (11)..(11) <223> Xaa is Ahp <400> 11 Ala Arg Xaa Tyr His Asp Arg Val Leu Xaa Xaa Asp Cys 1 5 10 <210> 12 <211> 13 <212> PRT <213> Synthetic Peptides <220> <221> MISC_FEATURE <222> (3) <223> Xaa is Ahp <220> <221> MISC_FEATURE <222> (7) <223> Xaa is Har <220> <221> MISC_FEATURE <222> (10)..(10) <223> Xaa is Bph <220> <221> MISC_FEATURE <222> (11)..(11) <223> Xaa is Ahp <400> 12 Ala Arg Xaa Tyr His Asp Xaa Val Leu Xaa Xaa Asp Cys 1 5 10 <210> 13 <211> 13 <212> PRT <213> Synthetic Peptides <220> <221> MISC_FEATURE <222> (3) <223> Xaa is Ahp <220> <221> MISC_FEATURE <222> (9)..(9) <223> Xaa is MetO2 <220> <221> MISC_FEATURE <222> (10)..(10) <223> Xaa is Bph <220> <221> MISC_FEATURE <222> (11)..(11) <223> Xaa is Ahp <400> 13 Ala Arg Xaa Tyr His Asp Gln Val Xaa Xaa Xaa Asp Cys 1 5 10 <210> 14 <211> 13 <212> PRT <213> Synthetic Peptides <220> <221> MISC_FEATURE <222> (3) <223> Xaa is Ahp <220> <221> MISC_FEATURE <222> (9)..(9) <223> Xaa is MetO2 <220> <221> MISC_FEATURE <222> (10)..(10) <223> Xaa is Bph <220> <221> MISC_FEATURE <222> (11)..(11) <223> Xaa is Ahp <400> 14 Ala Arg Xaa Tyr His Asp Glu Val Xaa Xaa Xaa Asp Cys 1 5 10 <210> 15 <211> 13 <212> PRT <213> Synthetic Peptides <220> <221> MISC_FEATURE <222> (3) <223> Xaa is Ahp <220> <221> MISC_FEATURE <222> (9)..(9) <223> Xaa is MetO2 <220> <221> MISC_FEATURE <222> (10)..(10) <223> Xaa is Bph <220> <221> MISC_FEATURE <222> (11)..(11) <223> Xaa is Ahp <400> 15 Ala Arg Xaa Tyr His Asp Arg Val Xaa Xaa Xaa Asp Cys 1 5 10 <210> 16 <211> 13 <212> PRT <213> Synthetic Peptides <220> <221> MISC_FEATURE <222> (3) <223> Xaa is Ahp <220> <221> MISC_FEATURE <222> (7) <223> Xaa is Har <220> <221> MISC_FEATURE <222> (9)..(9) <223> Xaa is MetO2 <220> <221> MISC_FEATURE <222> (10)..(10) <223> Xaa is Bph <220> <221> MISC_FEATURE <222> (11)..(11) <223> Xaa is Ahp <400> 16 Ala Arg Xaa Tyr His Asp Xaa Val Xaa Xaa Xaa Asp Cys 1 5 10 <210> 17 <211> 13 <212> PRT <213> Synthetic Peptides <220> <221> MISC_FEATURE <222> (1) <223> Xaa is Lys(MePEG4c) <220> <221> MISC_FEATURE <222> (3) <223> Xaa is Ahp <220> <221> MISC_FEATURE <222> (10)..(10) <223> Xaa is Bph <220> <221> MISC_FEATURE <222> (11)..(11) <223> Xaa is Ahp <400> 17 Xaa Arg Xaa Tyr His Asp Glu Val Leu Xaa Xaa Asp Cys 1 5 10 <210> 18 <211> 13 <212> PRT <213> Synthetic Peptides <220> <221> MISC_FEATURE <222> (2) <223> Xaa is Lys(MePEG4c) <220> <221> MISC_FEATURE <222> (3) <223> Xaa is Ahp <220> <221> MISC_FEATURE <222> (10)..(10) <223> Xaa is Bph <220> <221> MISC_FEATURE <222> (11)..(11) <223> Xaa is Ahp <400> 18 Ala Xaa Xaa Tyr His Asp Glu Val Leu Xaa Xaa Asp Cys 1 5 10 <210> 19 <211> 13 <212> PRT <213> Synthetic Peptides <220> <221> MISC_FEATURE <222> (3) <223> Xaa is Lys(MePEG4c) <220> <221> MISC_FEATURE <222> (10)..(10) <223> Xaa is Bph <220> <221> MISC_FEATURE <222> (11)..(11) <223> Xaa is Ahp <400> 19 Ala Arg Xaa Tyr His Asp Glu Val Leu Xaa Xaa Asp Cys 1 5 10 <210> 20 <211> 13 <212> PRT <213> Synthetic Peptides <220> <221> MISC_FEATURE <222> (1) <223> Xaa is da <220> <221> MISC_FEATURE <222> (3)..(3) <223> Xaa is Ahp <220> <221> MISC_FEATURE <222> (10)..(10) <223> Xaa is Bph <220> <221> MISC_FEATURE <222> (11)..(11) <223> Xaa is Ahp <400> 20 Xaa Arg Xaa Tyr His Asp Glu Val Leu Xaa Xaa Asp Cys 1 5 10 <210> 21 <211> 13 <212> PRT <213> Synthetic peptide <220> <221> MISC_FEATURE <222> (3)..(3) <223> Xaa is Ahp <220> <221> MISC_FEATURE <222> (10)..(10) <223> Xaa is Bph <220> <221> MISC_FEATURE <222> (11)..(11) <223> Xaa is Ahp <400> 21 Ala Arg Xaa Ala His Asp Glu Val Leu Xaa Xaa Asp Cys 1 5 10 <210> 22 <211> 13 <212> PRT <213> Synthetic Peptides <220> <221> MISC_FEATURE <222> (3) <223> Xaa is Ahp <220> <221> MISC_FEATURE <222> (5) <223> Xaa is W6N <220> <221> MISC_FEATURE <222> (10)..(10) <223> Xaa is Bph <220> <221> MISC_FEATURE <222> (11)..(11) <223> Xaa is Ahp <400> twenty two Ala Arg Xaa Tyr Xaa Asp Glu Val Leu Xaa Xaa Asp Cys 1 5 10 <210> twenty three <211> 13 <212> PRT <213> Synthetic Peptides <220> <221> MISC_FEATURE <222> (3) <223> Xaa is Ahp <220> <221> MISC_FEATURE <222> (10)..(10) <223> Xaa is Bph <220> <221> MISC_FEATURE <222> (11)..(11) <223> Xaa is Ano <400> twenty three Ala Arg Xaa Tyr His Asp Glu Val Leu Xaa Xaa Asp Cys 1 5 10 <210> twenty four <211> 13 <212> PRT <213> Synthetic Peptides <220> <221> MISC_FEATURE <222> (3) <223> Xaa is Ahp <220> <221> MISC_FEATURE <222> (10)..(10) <223> Xaa is Bph <220> <221> MISC_FEATURE <222> (11)..(11) <223> Xaa is Ado <400> twenty four Ala Arg Xaa Tyr His Asp Glu Val Leu Xaa Xaa Asp Cys 1 5 10 <210> 25 <211> 13 <212> PRT <213> Synthetic Peptides <220> <221> MISC_FEATURE <222> (3) <223> Xaa is Ahp <220> <221> MISC_FEATURE <222> (10)..(10) <223> Xaa is Bph <220> <221> MISC_FEATURE <222> (11)..(11) <223> Xaa is PhNle <400> 25 Ala Arg Xaa Tyr His Asp Glu Val Leu Xaa Xaa Asp Cys 1 5 10 <210> 26 <211> 13 <212> PRT <213> Synthetic Peptides <220> <221> MISC_FEATURE <222> (3) <223> Xaa is Ahp <220> <221> MISC_FEATURE <222> (10)..(10) <223> Xaa is Bph <220> <221> MISC_FEATURE <222> (11)..(11) <223> Xaa is PhNva <400> 26 Ala Arg Xaa Tyr His Asp Glu Val Leu Xaa Xaa Asp Cys 1 5 10 <210> 27 <211> 13 <212> PRT <213> Synthetic Peptides <220> <221> MISC_FEATURE <222> (3) <223> Xaa is Ahp <220> <221> MISC_FEATURE <222> (10)..(10) <223> Xaa is Bph <220> <221> MISC_FEATURE <222> (11)..(11) <223> Xaa is Ahp <400> 27 Ala Arg Xaa Tyr His Asp Glu Val Leu Xaa Xaa Ala Cys 1 5 10 <210> 28 <211> 13 <212> PRT <213> Synthetic Peptides <220> <221> MISC_FEATURE <222> (2) <223> Xaa is Cit <220> <221> MISC_FEATURE <222> (3) <223> Xaa is Ahp <220> <221> MISC_FEATURE <222> (10)..(10) <223> Xaa is Bph <220> <221> MISC_FEATURE <222> (11)..(11) <223> Xaa is Ahp <400> 28 Ala Xaa Xaa Tyr His Asp Glu Val Leu Xaa Xaa Asp Cys 1 5 10 <210> 29 <211> 13 <212> PRT <213> Synthetic Peptides <220> <221> MISC_FEATURE <222> (2) <223> Xaa is F3G <220> <221> MISC_FEATURE <222> (3) <223> Xaa is Ahp <220> <221> MISC_FEATURE <222> (10)..(10) <223> Xaa is Bph <220> <221> MISC_FEATURE <222> (11)..(11) <223> Xaa is Ahp <400> 29 Ala Xaa Xaa Tyr His Asp Glu Val Leu Xaa Xaa Asp Cys 1 5 10 <210> 30 <211> 13 <212> PRT <213> Synthetic peptide <220> <221> MISC_FEATURE <222> (2)..(2) <223> Xaa is RNMe <220> <221> MISC_FEATURE <222> (3)..(3) <223> Xaa is Ahp <220> <221> MISC_FEATURE <222> (10)..(10) <223> Xaa is Bph <220> <221> MISC_FEATURE <222> (11)..(11) <223> Xaa is Ahp <400> 30 Ala Xaa Xaa Tyr His Asp Glu Val Leu Xaa Xaa Asp Cys 1 5 10 <210> 31 <211> 13 <212> PRT <213> Synthetic peptide <220> <221> MISC_FEATURE <222> (2) <223> Xaa is RNNdMe <220> <221> MISC_FEATURE <222> (3) <223> Xaa is Ahp <220> <221> MISC_FEATURE <222> (10)..(10) <223> Xaa is Bph <220> <221> MISC_FEATURE <222> (11)..(11) <223> Xaa is Ahp <400> 31 Ala Xaa Xaa Tyr His Asp Glu Val Leu Xaa Xaa Asp Cys 1 5 10 <210> 32 <211> 13 <212> PRT <213> Synthetic Peptides <220> <221> MISC_FEATURE <222> (2) <223> Xaa is RNdMe <220> <221> MISC_FEATURE <222> (3) <223> Xaa is Ahp <220> <221> MISC_FEATURE <222> (10)..(10) <223> Xaa is Bph <220> <221> MISC_FEATURE <222> (11)..(11) <223> Xaa is Ahp <400> 32 Ala Xaa Xaa Tyr His Asp Glu Val Leu Xaa Xaa Asp Cys 1 5 10 <210> 33 <211> 13 <212> PRT <213> Synthetic Peptides <220> <221> MISC_FEATURE <222> (2) <223> Xaa is hCit <220> <221> MISC_FEATURE <222> (3) <223> Xaa is Ahp <220> <221> MISC_FEATURE <222> (10)..(10) <223> Xaa is Bph <220> <221> MISC_FEATURE <222> (11)..(11) <223> Xaa is Ahp <400> 33 Ala Xaa Xaa Tyr His Asp Glu Val Leu Xaa Xaa Asp Cys 1 5 10 <210> 34 <211> 13 <212> PRT <213> Synthetic Peptides <220> <221> MISC_FEATURE <222> (2) <223> Xaa is 4Py2NH2 <220> <221> MISC_FEATURE <222> (3) <223> Xaa is Ahp <220> <221> MISC_FEATURE <222> (10)..(10) <223> Xaa is Bph <220> <221> MISC_FEATURE <222> (11)..(11) <223> Xaa is Ahp <400> 34 Ala Xaa Xaa Tyr His Asp Glu Val Leu Xaa Xaa Asp Cys 1 5 10

Claims

1. A CD38 binding peptide, a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable solvate thereof, characterized in that: The CD38 binding peptide is a peptide having a cyclized structure and consisting of an amino acid sequence represented by SEQ ID NO. 1 or 2: Ala Arg Ahp Tyr His Asp Gly Val Leu Bph Ahp Asp Cys (SEQ ID NO.1), Ala Leu His MePhe Val Leu Pro Bph Val Trp Val Cys (SEQ ID NO. 2); The peptide having a cyclized structure and consisting of the amino acid sequence represented by SEQ ID NO.1 has the following chemical formula: ， The peptide having a cyclized structure and consisting of the amino acid sequence represented by SEQ ID NO.2 has the following chemical formula: 。 2. A compound comprising the CD38 binding peptide according to claim 1, a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable solvate thereof, characterized in that: The CD38 binding peptide, its pharmaceutically acceptable salt or its pharmaceutically acceptable solvate further comprises a linking moiety.

3. The compound according to claim 2, characterized in that The linking moiety comprises polyethylene glycol.

4. A pharmaceutical composition for treating CD38-related cancer, characterized in that: Include: A CD38 binding peptide according to claim 1, a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable solvate thereof; and A pharmaceutically acceptable carrier.

5. The pharmaceutical composition according to claim 4, characterized in that The CD38-related cancer is leukemia or myeloma.

6. The pharmaceutical composition according to claim 4, characterized in that The CD38-related cancer is B-cell non-Hodgkin's lymphoma, multiple myeloma (MM), acute myeloid leukemia (AML), acute lymphocytic leukemia (ALL) or chronic lymphocytic leukemia (CLL).

7. The pharmaceutical composition according to claim 4, characterized in that The CD38-related cancer is multiple myeloma (MM).

8. A cancer detection reagent, characterized in that: The reagent comprises the CD38-binding peptide according to claim 1, or a pharmaceutically acceptable salt or a pharmaceutically acceptable solvate thereof.

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