Compositions for targeted delivery to adipose tissue

Abstract

The present disclosure includes, inter alia, compositions comprising cyclic peptides selective for adipose tissue. The present disclosure includes, inter alia, the recognition that such cyclic peptides selective for adipose tissue can be used to target the delivery of an effective payload (e.g., an antisense oligonucleotide or small interfering RNA) to adipose tissue by conjugating such peptides to the payload. The present disclosure includes, inter alia, compositions comprising cyclic peptide conjugates and their use in treating disorders of adipose tissue.

Description

Cross-references to related applications

[0001] This application claims the benefit of priority to U.S. Provisional Application No. 63 / 550,783, filed February 7, 2024, the contents of which are incorporated herein by reference in their entirety. Background Technology

[0002] Adipose tissue is an important metabolic and endocrine organ that contributes to biological functions, including energy storage and energy homeostasis. Impairment of adipose tissue function is associated with a variety of diseases. Delivering drugs to adipose tissue can be used to treat these diseases. Summary of the Invention

[0003] This disclosure specifically provides compositions comprising cyclic peptides targeting adipose tissue, and the use of such cyclic peptides in the treatment of diseases. Not wishing to be bound by any particular scientific theory, this disclosure includes the unexpected finding that the cyclic peptides disclosed herein can selectively target receptors present in adipose tissue. Therefore, this disclosure specifically provides compositions of cyclic peptides selectively targeting adipose tissue, conjugates comprising the cyclic peptides of this disclosure and therapeutic or diagnostic agents, pharmaceutical compositions, and the use of the pharmaceutical agents disclosed herein for the treatment of diseases (e.g., diseases related to adipose tissue).

[0004] Therefore, this disclosure specifically provides conjugates comprising the cyclic peptides disclosed herein associated with therapeutic or diagnostic agents, and the use of such conjugates in the treatment of diseases. Without wishing to be bound by any particular scientific theory, this disclosure is based, at least in part, on the understanding that conjugates comprising cyclic peptides can selectively deliver associated therapeutic or diagnostic agents to adipose tissue. As disclosed herein, the cyclic peptides of this disclosure can be associated with therapeutic or diagnostic agents, which can be, but are not limited to, nucleic acids, peptides, or chemical agents. Nucleic acid agents can be, for example, repressive nucleic acids or nucleic acids encoding expression products (e.g., transgenes). Repressive nucleic acids can be selected from, but are not limited to, small interfering RNA (siRNA), microRNA (miRNA), and repressive antisense oligonucleotides (ASO).

[0005] In at least one aspect, this disclosure provides a composition comprising a cyclic peptide selectively targeting adipose tissue, wherein the cyclic peptide comprises an amino acid sequence differing from a sequence selected from SEQ ID NO: 1-161 by no more than three amino acids, wherein each amino acid difference is independently selected from amino acid insertion, deletion, or substitution. In some embodiments, the cyclic peptide comprises an amino acid sequence selected from SEQ ID NO: 1-161. In some embodiments, the cyclic peptide comprises an amino acid sequence differing from a sequence selected from SEQ ID NO: 1-3. In some embodiments, the cyclic peptide comprises an amino acid sequence selected from SEQ ID NO: 1-3. In some embodiments, the cyclic peptide comprises an amino acid sequence differing from SEQ ID NO: 1 by no more than three amino acids. In some embodiments, the cyclic peptide comprises an amino acid sequence according to SEQ ID NO: 1. In some embodiments, the cyclic peptide comprises an amino acid sequence differing from SEQ ID NO: 2 by no more than three amino acids. In some embodiments, the cyclic peptide comprises an amino acid sequence according to SEQ ID NO: 2. In some embodiments, the cyclic peptide comprises an amino acid sequence that differs from SEQ ID NO: 3 by no more than three amino acids. In some embodiments, the cyclic peptide comprises the amino acid sequence according to SEQ ID NO: 3. In some embodiments, the adipose tissue is white adipose tissue (WAT).

[0006] In at least one aspect, this disclosure provides a peptide conjugate comprising a cyclic peptide and a pharmaceutical agent associated with the cyclic peptide, the cyclic peptide comprising an amino acid sequence differing from a sequence selected from SEQ ID NO: 1-161 by no more than three amino acids, wherein each amino acid difference is independently selected from an amino acid insertion, deletion, or substitution. In some embodiments, the cyclic peptide comprises an amino acid sequence selected from SEQ ID NO: 1-161. In some embodiments, the cyclic peptide comprises an amino acid sequence differing from a sequence selected from SEQ ID NO: 1-3 by no more than three amino acids. In some embodiments, the cyclic peptide comprises an amino acid sequence selected from SEQ ID NO: 1-3. In some embodiments, the cyclic peptide comprises an amino acid sequence differing from SEQ ID NO: 1 by no more than three amino acids. In some embodiments, the cyclic peptide comprises an amino acid sequence according to SEQ ID NO: 1. In some embodiments, the cyclic peptide comprises an amino acid sequence differing from SEQ ID NO: 2 by no more than three amino acids. In some embodiments, the cyclic peptide comprises an amino acid sequence according to SEQ ID NO: 2. In some embodiments, the cyclic peptide comprises an amino acid sequence that differs from SEQ ID NO: 3 by no more than three amino acids. In some embodiments, the cyclic peptide comprises the amino acid sequence according to SEQ ID NO: 3.

[0007] In various embodiments, the agent is a diagnostic or therapeutic agent. In some embodiments, the therapeutic agent targets targets according to Table 2. In some embodiments, the agent is an inhibitory nucleic acid. In some embodiments, the agent is an antisense oligonucleotide. In some embodiments, the agent is siRNA or miRNA. In some embodiments, the antisense oligonucleotide is a phosphodiamide morpholino oligonucleotide (PMO) or peptide nucleic acid (PNA). In some embodiments, the agent is adeno-associated virus (AAV). In some embodiments, the agent is lipid nanoparticles (LNP). In some embodiments, the agent is non-covalently associated with the cyclic peptide. In some embodiments, the agent is covalently associated with the cyclic peptide. In some embodiments, the agent is directly covalently associated with the cyclic peptide. In some embodiments, the agent is indirectly covalently associated with the cyclic peptide. In some embodiments, the agent is indirectly covalently associated with the cyclic peptide via a linker. In some embodiments, the linker includes a thioether bond, a disulfide bond, an oxime, a thiazolidinyl ether, a hydrazone, an amide bond, an azide bond, or a maleimide bond. In some implementations, the connector includes C1-C 30 Alkyl, C2-C 20 Alkyl, C3-C 12 Alkyl, C6-C 12Alkyl or C6 alkyl group. In some embodiments, the connector includes a C6 alkylamine. In some embodiments, the connector is a cuttable connector.

[0008] In at least one aspect, this disclosure provides a pharmaceutical composition comprising the peptide conjugate disclosed herein and a pharmaceutically acceptable carrier. In at least one aspect, this disclosure provides a method of delivering a pharmaceutical agent to the adipose tissue of a subject, the method comprising administering to the subject the peptide conjugate or the pharmaceutical composition disclosed herein. In at least one aspect, this disclosure provides a method for diagnosing, preventing, and / or treating adipose tissue-related diseases, the method comprising administering to a subject the peptide conjugate or the pharmaceutical composition disclosed herein. In some embodiments, the disease is selected from obesity, cachexia, hyperglycemia, insulin resistance, type 2 diabetes, hypertension, cancer, heart disease, immune diseases, arthritis, diseases of the central nervous system, metabolic disorders, and age-related diseases. Attached Figure Description

[0009] Figure 1 This is a graph showing the expression and knockdown of Malat1 after delivery of an exemplary antisense oligonucleotide (ASO)-cyclic peptide conjugate. Figure 1 This study provides quantification of Malat1 expression in white adipose tissue (WAT) in mice following in vivo delivery of ASOs conjugated with three exemplary targeting cyclic peptides having the amino acid sequences of SEQ ID NO: 1-3, respectively. ASOs conjugated with control peptides were delivered as a non-targeting control. Expression was normalized to Malat1 expression levels in animals administered PBS.

[0010] Figure 2 This is a graph showing the expression and knockdown of Malat1 after delivery of an exemplary antisense oligonucleotide (ASO)-cyclic peptide conjugate. Figure 2 Quantification of Malat1 knockdown induced in various tissues by exemplary ASO-cyclic peptide conjugates relative to non-targeted controls is provided.

[0011] Figure 3 This is shown after delivery of an exemplary small interfering RNA (siRNA)-cyclic peptide conjugate at 5 mg / kg. ALDH2 The expression and the low-key image. Figure 3 Provides the ability to deliver siRNA into designated tissues in mice after subcutaneous delivery. ALDH2 Quantification of the expression of the targeted gene. The siRNA conjugate contains a targeting cyclic peptide having the amino acid sequence SEQ ID NO: 3. For each tissue shown on the X-axis, data include administration of either the PBS control (left) or the conjugated cyclic peptide (right). Expression was normalized to the expression of the target gene in animals administered PBS. Detailed Implementation

[0012] This disclosure specifically provides compositions comprising cyclic peptides that selectively target adipose tissue, conjugates of cyclic peptides with therapeutic or diagnostic agents, and uses of such peptides and conjugates for delivering pharmaceutical agents to adipose tissue (e.g., for treating or diagnosing a disease). The cyclic peptides of this disclosure can associate with a variety of pharmaceutical agents, including but not limited to nucleic acids, peptides, or chemical agents. Nucleic acid agents can be, for example, repressive nucleic acids or nucleic acids encoding expressed products (e.g., transgenes). Repressive nucleic acids can be selected from, but are not limited to, repressive antisense oligonucleotides (ASOs), small interfering RNA (siRNA), and microRNA (miRNA).

[0013] definition A / an, the As used herein, “a / an” and “the” refer to one or more (i.e., at least one) grammatical object of the article. For example, “an element” discloses an implementation of exactly one element as well as an implementation that includes more than one element.

[0014] about: As used herein, the term "about" when used to refer to a value means a value similar to the reference value in the context. Generally, those skilled in the art will understand the extent of variation covered by "about" in this context. For example, in some embodiments, the term "about" may cover a range of values ​​within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less of the reference value.

[0015] acyl group The term "acyl" is a technical term and, as used herein, refers to any group or atomic group of the form RCO-, where R is any organic group, such as alkyl, aryl, heteroaryl, aralkyl, and heteroaryl. Representative acyl groups include acetyl, benzoyl, and malonyl.

[0016] Administration (or administering) As used herein, the term “administration” generally refers to the administration of a composition to a subject or system to achieve delivery of a pharmaceutical agent as a composition or contained in a composition.

[0017] Alkenoxy (alkenoxy or alkenoxyl) The term "olefinic group" means, as defined herein, an alkenyl group attached to a portion of a parent molecule by an oxygen atom. Representative examples of olefinic groups include, but are not limited to, 2-propen-1-oxy (i.e., CH2=CH-CH2-O-) and vinyloxy (i.e., CH2=CH-O-).

[0018] Alkoxy As used herein, the term "alkoxy" means an alkyl group attached to a portion of a parent molecule by an oxygen atom, as defined herein. Representative examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, 2-propoxy, butoxy, tert-butoxy, pentoxy, and hexoxy.

[0019] alkoxycarbonyl The term "alkoxycarbonyl" means, as defined herein, an alkoxy group attached to a portion of a parent molecule by a carbonyl group represented by -C(=O)- as defined herein. Representative examples of alkoxycarbonyl include, but are not limited to, methoxycarbonyl, ethoxycarbonyl, and tert-butoxycarbonyl.

[0020] alkyl carbonyl As used herein, the term "alkyl carbonyl" means an alkyl group attached to a portion of a parent molecule by a carbonyl group as defined herein. Representative examples of alkyl carbonyl groups include, but are not limited to, acetyl, 1-oxopropyl, 2,2-dimethyl-1-oxopropyl, 1-oxobutyl, and 1-oxopentyl.

[0021] alkyl carbonyloxy or aryl carbonyloxy As used herein, the terms "alkylcarbonyloxy" and "arylcarbonyloxy" mean an alkylcarbonyl or arylcarbonyl group attached to a portion of a parent molecule by an oxygen atom as defined herein. Representative examples of alkylcarbonyloxy groups include, but are not limited to, acetoxy, ethylcarbonyloxy, and tert-butylcarbonyloxy. Representative examples of arylcarbonyloxy groups include, but are not limited to, phenylcarbonyloxy.

[0022] Aryl or arylalkyl The terms “aralkyl” or “arylalkyl” are technical terms and, as used herein, refer to alkyl groups that are substituted with aryl groups, some of which are attached to the parent molecule via the alkyl group.

[0023] aryl carbonyl As used herein, the term "aryl carbonyl" means an aryl group attached to a parent molecule moiety by a carbonyl group as defined herein. Representative examples of aryl carbonyl groups include, but are not limited to, benzoyl and (2-pyridyl)carbonyl.

[0024] aryloxy As used herein, the term "aryloxy group" means, as defined herein, an aryl group attached to a portion of the parent molecule by an oxygen atom.

[0025] medicineAs used herein, the term "pharmaceutical" may refer to any chemical entity, including but not limited to atoms, molecules, compounds, amino acids, polypeptides, nucleotides, nucleic acids, proteins, protein complexes, liquids, solutions, sugars, polysaccharides, lipids, or combinations or complexes thereof.

[0026] alkyl As used herein, the term "alkyl" is a technical term and refers to a saturated aliphatic group, including straight-chain alkyl groups, branched alkyl groups, cycloalkyl (alicyclic) groups, alkyl-substituted cycloalkyl groups, and cycloalkyl-substituted alkyl groups. In some embodiments, the straight-chain or branched alkyl group has 30 or fewer carbon atoms in its main chain (e.g., for a straight chain, C1-C1). 30 For sidechains, C3-C 30 And alternatively, about 20 or fewer, or 10 or fewer carbon atoms. In some embodiments, the term "alkyl" refers to C1-C2. 10 Straight-chain alkyl groups. In some embodiments, the term "alkyl" refers to a C1-C6 straight-chain alkyl group. In some embodiments, the term "alkyl" refers to a C3-C6 straight-chain alkyl group. 12 Branched alkyl groups. In some embodiments, the term "alkyl" refers to a C3-C8 branched alkyl group. Representative examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, and n-hexyl.

[0027] alkenyl As used herein, the term "alkenyl" refers to a straight-chain or branched hydrocarbon group containing 2 to 10 carbons and at least one carbon-carbon double bond formed by the removal of two hydrogens. Representative examples of alkenyl groups include, but are not limited to, vinyl, 2-propenyl, 2-methyl-2-propenyl, 3-butenyl, 4-pentenyl, 5-hexenyl, 2-heptenyl, 2-methyl-1-heptenyl, and 3-decenyl. One or more unsaturated bonds in the alkenyl group can be located at any position within the moiety and can have a (Z) or (E) configuration surrounding one or more double bonds.

[0028] AlkyleneThe term "alkylene" is recognized in the art and, as used herein, refers to a bimolecular group obtained by removing two hydrogen atoms from an alkyl group as defined above. In one embodiment, alkylene refers to a disubstituted alkane, i.e., an alkane in which two positions are substituted by substituents such as halogen, azide, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, alkoxy, amino, nitro, mercapto, imino, amide, phosphonate, phosphonite, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, sulfonamide, ketone, aldehyde, ester, heterocyclic, aromatic or heteroaromatic moiety, fluoroalkyl (such as trifluoromethyl), cyano, etc. That is, in one embodiment, "substituted alkyl" is "alkylene".

[0029] alkylphosphoyl As used herein, the term "alkylphosphoryl" means a phosphoryl group substituted with at least one alkyl group as defined herein; for example, -P(O)(OH)Me.

[0030] Alkylthio As used herein, the term "alkylthio" refers to alkyl-S-.

[0031] acetylin As used herein, the term "alkynyl" refers to a straight-chain or branched hydrocarbon group containing 2 to 10 carbon atoms and at least one carbon-carbon triple bond. Representative examples of alkynyl groups include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, 3-butynyl, 2-pentynyl, and 1-butynyl.

[0032] Acylamino As used herein, the term "acylamino" refers to -NHC(=O)-, where the acylamino group is attached to the parent molecule via nitrogen. Examples of acylamino groups include alkyl acylamino groups, such as CH3C(=O)N(H)- and CH3CH2C(=O)N(H)-.

[0033] amino The term "amino" is a technical term and, as used herein, refers to both unsubstituted and substituted amines, for example, portions that can be represented by the following general formula: and , Where R a R b and R c Each can independently represent hydrogen, alkyl, alkenyl, or -(CH2). x -R d , or R a and R b Together with the attached N atom, it completes a heterocycle with 4 to 8 atoms in the ring structure; R dRepresents an aryl, cycloalkyl, cycloalkenyl, heterocyclic, or polycyclic group; and x is zero or an integer in the range of 1 to 8. In some embodiments, R a Or R b Only one of them can be a carbonyl group, such as R. a R b It does not form an imide with nitrogen. In other embodiments, R a and R b (and optional R) c Each can independently represent hydrogen, alkyl, alkenyl, or -(CH2). x -R d In some implementations, the term "amino" refers to –NH2.

[0034] Amino acids: As used herein, the term "amino acid" in its broadest sense refers to any compound and / or substance that can, for example, bind to a polypeptide chain by forming one or more peptide bonds. In some embodiments, the amino acid is a naturally occurring amino acid. In some embodiments, the amino acid is a non-natural amino acid; in some embodiments, the amino acid is a D-amino acid; in some embodiments, the amino acid is an L-amino acid. "Standard amino acid" refers to any of the twenty standard L-amino acids that are typically present in naturally occurring peptides. "Non-standard amino acid" refers to any amino acid other than a standard amino acid, whether that amino acid is synthetically prepared or obtained from a natural source. In some embodiments, amino acids (including carboxyl-terminal and / or amino-terminal amino acids in polypeptides) may contain structural modifications compared to a typical or canonical amino acid structure. For example, in some embodiments, amino acids may be modified by methylation, amidation, acetylation, polyethylene glycolation, glycosylation, phosphorylation, sulfation, and / or substitution (e.g., substitution of an amino group, a carboxylic acid group, one or more protons, and / or hydroxyl groups) compared to a general structure. In some embodiments, such modification can, for example, alter the cyclic half-life of a polypeptide containing modified amino acids compared to a polypeptide containing otherwise identical, unmodified amino acids. In some embodiments, such modification does not significantly alter the relevant activity of a polypeptide containing modified amino acids compared to a polypeptide containing otherwise identical, unmodified amino acids. As will be clear from the context, in some embodiments, the term "amino acid" can be used to refer to a free amino acid; in some embodiments, it can be used to refer to an amino acid residue of a polypeptide. In the context of any amino acid present in a polypeptide, or when any amino acid is considered or discussed in the context of its presence or potential presence in a polypeptide, as used herein, the terms amino acid and residue are interchangeable.

[0035] aminoacylThe term "aminoacyl" is a technical term and, as used herein, refers to an acyl group that is substituted with one or more amino groups.

[0036] aminoalkyl As used herein, the term "aminoalkyl" refers to an alkyl group substituted with one or more amino groups. In one embodiment, the term "aminoalkyl" refers to an aminomethyl group.

[0037] aminophosphoyl As used herein, the term "aminophosphoryl" refers to a phosphoryl group substituted with at least one amino group as defined herein; for example, -P(O)(OH)NMe2.

[0038] aminothionyl As used herein, the term "aminothionyl" refers to an analog of an aminoacyl group in which the O in RC(O)- has been replaced by sulfur, thus presenting as RC(S)-.

[0039] Aryl The term "aryl" is a technical term and, as used herein, refers to a group comprising monocyclic, bicyclic, and polycyclic aromatic hydrocarbon groups, such as benzene, naphthalene, anthracene, and pyrene. An aromatic ring may be substituted at one or more ring positions with one or more substituents, such as halogens, azides, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, alkoxy, amino, nitro, mercapto, imino, amide, phosphonate, phosphonite, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, sulfonamide, ketone, aldehyde, ester, heterocyclic, aromatic or heteroaromatic moiety, fluoroalkyl (such as trifluoromethyl), cyano, etc. The term "aryl" also includes polycyclic ring systems having two or more cyclic rings, wherein two or more carbons are common to two adjacent rings (the rings are "fused rings"), wherein at least one ring is an aromatic hydrocarbon, and other cyclic rings may be cycloalkyl, cycloalkenyl, cycloynyl, aryl, heteroaryl, and / or heterocyclic. In some embodiments, the term "aryl" refers to a phenyl group. In some embodiments, "aryl" has 6 to 10 carbon atoms.

[0040] Associated with:Two events or entities are “related” to each other, as used herein, if the presence, level, and / or form of one event or entity is related to the presence, level, and / or form of another event or entity. For example, if the presence, level, and / or form of a particular entity (e.g., polypeptide, genetic marker, metabolite, microorganism, etc.) is related to the incidence and / or susceptibility to a disease, symptom, or condition (e.g., in a relevant population), then that particular entity is considered related to that particular disease, symptom, or condition. In some embodiments, two or more entities are physically “associated” to each other if they interact directly or indirectly (e.g., covalently or non-covalently) such that they are physically close to each other and / or remain physically close. In some embodiments, two or more entities physically associated to each other are covalently linked; in some embodiments, two or more entities physically associated to each other are not covalently linked but non-covalently associated, for example, by means of hydrogen bonds, van der Waals interactions, hydrophobic interactions, magnetism, or combinations thereof.

[0041] azide or azide group As used herein, the term “azide” or “azido group” refers to the –N3 group.

[0042] Between or from As used herein, the term “between” means the content between the indicated upper and lower limits or between the first and second boundaries (or “limits”), including the boundaries. Similarly, the term “from” when used in the context of a range of values ​​means that the range includes the content between the indicated upper and lower limits or between the first and second boundaries (inclusive).

[0043] biological samplesAs used herein, the term "biological sample" generally refers to a sample obtained or derived from a biological source of interest (e.g., a tissue or organism or cell culture), as described herein. In some embodiments, the biological source is or includes an organism, such as an animal or a human. In some embodiments, the biological sample is or includes biological tissue or fluid. In some embodiments, the biological sample may be or includes cells, tissues, or body fluids. In some embodiments, the biological sample may be or includes blood, blood cells, cell-free DNA, cell-free nucleic acids, ascites, biopsy samples, surgical samples, cellular body fluids, sputum, saliva, feces, urine, cerebrospinal fluid, peritoneal fluid, pleural fluid, lymph, gynecological fluids, secretions, excretions, skin swabs, vaginal swabs, oral swabs, nasal swabs, washing or lavage solutions (such as catheter lavage solution or bronchoalveolar lavage solution), aspirates, scrapings, or bone marrow. In some embodiments, the biological sample is or includes cells obtained from a single subject or from multiple subjects. A sample can be a “raw sample” obtained directly from a biological source, or it can be a “processed sample” (e.g., a sample prepared from a raw sample, for example by processes such as isolating mRNA, DNA, or proteins, by altering the chemical structure of the raw sample, and / or by producing a new or different composition that represents one or more components or properties of the raw sample). Biological samples can also be referred to as “samples”.

[0044] carbon cyclo group As used herein, the term "carbocyclic" refers to a monocyclic or polycyclic (e.g., bicyclic, tricyclic, etc.) hydrocarbon group containing 3 to 12 carbon atoms, which is either fully saturated or has one or more unsaturated bonds, and for the avoidance of doubt, the degree of unsaturation does not result in an aromatic ring system (e.g., phenyl). Examples of carbocyclic groups include 1-cyclopropyl, 1-cyclobutyl, 2-cyclopentyl, 1-cyclopentenyl, 3-cyclohexyl, 1-cyclohexenyl, and 2-cyclopentenylmethyl.

[0045] carbonyl As used in this article, the term "carbonyl" refers to -C(=O)-.

[0046] carboxyl As used in this article, the term "carboxyl" refers to the -CO2H group.

[0047] Conservative replacement:A conservative substitution is a substitution in which an amino acid has been replaced by a non-identical residue having a suitably similar structural and / or functional characteristic. For example, as is well known to those skilled in the art, certain amino acids are generally classified as "hydrophobic" or "hydrophilic" amino acids, and / or have "polar" or "nonpolar" side chains. The substitution of one amino acid for another of the same type can generally be considered a conservative substitution. A summary of some non-restrictive amino acid classifications is provided below. Those skilled in the art will understand the classifications and relationships that can be used to identify conservative substitutions among amino acids.

[0048] cycloalkyl The term "cycloalkyl" refers to a monocyclic, bicyclic, or bridged saturated carbocyclic ring, each having 3 to 12 carbon atoms. Some cycloalkyl groups have 5-12 carbon atoms in their ring structure, and some may have 6-10 carbon atoms. Preferably, the cycloalkyl group is a (C3-C7)cycloalkyl group, representing a monocyclic saturated carbocyclic ring having 3 to 7 carbon atoms. Examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, and cyclooctyl. Bicyclic cycloalkyl ring systems include bridged monocyclic rings and fused bicyclic rings. Bridged monocyclic rings contain a monocyclic cycloalkyl ring, wherein two non-adjacent carbon atoms of the monocyclic ring are connected by an alkylene bridge between one to three additional carbon atoms (i.e., in the form -(CH2)). w - bridging groups, wherein w (1, 2, or 3). Representative examples of bicyclic ring systems include, but are not limited to, bicyclic [3.1.1]heptane, bicyclic [2.2.1]heptane, bicyclic [2.2.2]octane, bicyclic [3.2.2]nonane, bicyclic [3.3.1]nonane, and bicyclic [4.2.1]nonane. Fused bicyclic cycloalkyl ring systems contain a monocyclic cycloalkyl ring fused with a phenyl, monocyclic cycloalkyl, monocyclic cycloalkenyl, monocyclic heterocyclic, or monocyclic heteroaryl group. The bridged or fused bicyclic cycloalkyl group is attached to the parent molecule moiety via any carbon atom contained within the monocyclic cycloalkyl ring. The cycloalkyl group is optionally substituted. In some embodiments, the fused bicyclic cycloalkyl group is a 5 or 6-membered monocyclic cycloalkyl ring fused to a benzene ring, a 5 or 6-membered monocyclic cycloalkyl group, a 5 or 6-membered monocyclic cycloalkenyl group, a 5 or 6-membered monocyclic heterocyclic group, or a 5 or 6-membered monocyclic heteroaryl group, wherein the fused bicyclic cycloalkyl group is optionally substituted.

[0049] cyano The term "cyano" is a technical term and, as used in this article, refers to –CN.

[0050] cycloalkyl alkyl As used herein, the term "cycloalkylalkyl" refers to an alkyl group that is substituted by one or more cycloalkyl groups. An example of a cycloalkylalkyl group is a cyclohexylmethyl group.

[0051] Gene As used herein, the term “gene” refers to a DNA sequence that is or contains a coding sequence (i.e., a DNA sequence that encodes an expression product, such as an RNA product and / or a polypeptide product), optionally together with some or all of the regulatory sequences that control the expression of the coding sequence.

[0052] Express As used herein, “expression” refers, individually and / or cumulatively, to one or more biological processes that result in the production of an agent encoded by a nucleic acid sequence (i.e., an expression product), such as RNA and / or a polypeptide. Expression specifically includes either or both of transcription and translation. The nucleic acid or cell that produces the encoded agent may be referred to as expressing the agent encoded by it.

[0053] Halogenated group The term “halogenated group” is a technical term and, as used herein, refers to –F, –Cl, –Br, or –I.

[0054] Halogenated alkyl As used herein, the term “halogenated alkyl” refers to an alkyl group in which some or all of its hydrogen atoms are replaced by halogen atoms, as defined herein.

[0055] heteroarylThe term “heteroaryl” is a technical term and, as used herein, refers to a monocyclic, bicyclic, or polycyclic aromatic group having 3 to 12 total atoms (including one or more heteroatoms such as nitrogen, oxygen, or sulfur) in a ring structure. Exemplary heteroaryl groups include azaindolyl, benzo(b)thiophenyl, benzimidazolyl, benzofuranyl, benzooxazolyl, benzothiazolyl, benzothiadiazolyl, benzotriazolyl, benzooxadiazolyl, furanyl, imidazolyl, imidazopyridyl, indolyl, dihydroindolyl, indazole, isodihydroindolyl, isoxazolyl, isothiazolyl, isoquinolinyl, oxadiazolyl, oxazolyl, purinyl, pyranyl, pyrazinyl, pyrazolyl, pyridyl, pyrimidinyl, pyrroloyl, pyrrolo[2,3-d]pyrimidinyl, pyrazolo[3,4-d]pyrimidinyl, quinolinyl, quinazolinyl, triazolyl, thiazolyl, thiophenyl, tetrahydroindolyl, tetraazolyl, thiadiazolyl, thiophenyl, thiomorpholinyl, triazolyl, or scopolamine, etc. "Heteroaryl" can be substituted at one or more ring positions by one or more substituents, such as halogens, azides, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, alkoxy, amino, nitro, mercapto, imino, amide, phosphonate, phosphonite, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, sulfonamide, ketone, aldehyde, ester, heterocyclic group, aromatic or heteroaromatic moiety, fluoroalkyl (such as trifluoromethyl), cyano, etc. The term "heteroaryl" also includes polycyclic ring systems having two or more cyclic rings, wherein two or more carbons are common to two adjacent rings (the ring is a "fused ring"), wherein at least one ring is an aromatic group having one or more heteroatoms in the ring structure; for example, other cyclic rings can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclic groups.

[0056] heteroaryl or heteroarylalkyl The terms “heteroarylalkyl” or “heteroarylalkyl” are technical terms and, as used herein, refer to an alkyl group substituted with a heteroaryl group that is attached to the parent molecule via an alkyl group.

[0057] heteroaryloxy As used herein, the term “heteroaryloxy” means a heteroaryl group attached to a portion of the parent molecule by an oxygen atom, as defined herein.

[0058] heteroatoms The term "heteroatom" is generally accepted in the art and refers to an atom of any element other than carbon or hydrogen. Illustrative heteroatoms include boron, nitrogen, oxygen, phosphorus, sulfur, and selenium, or oxygen, nitrogen, or sulfur.

[0059] heterocyclic groupAs used herein, the term "heterocyclic group" refers to a group of a non-aromatic ring system (including, but not limited to, monocyclic, bicyclic, and tricyclic rings) that may be fully saturated or may contain one or more unsaturated units. For the avoidance of doubt, the unsaturation does not produce an aromatic ring system and has 3 to 12 atoms, including at least one heteroatom such as nitrogen, oxygen, or sulfur. For illustrative purposes and not to be construed as limiting the scope of the invention, the following are examples of heterocyclic rings: aziridinyl, acryloxy, ethylene oxide, cyclothioethylene, thiapropenyl, dicyclooxyethylene, diazacyclopropenyl, diazacycloheptyl, 1,3-dioxane, 1,3-dioxopentyl, 1,3-dithiopentanyl, 1,3-dithiaalkyl, imidazoalkyl, isothiazolinyl, isothiazolinyl, isoxazolinyl, azetyl, oxacyclobutadiene, oxacyclobutadiene, thiobutenyl, thiacyclobutadiene, diazacyclobutane, dioxacyclobutane, dioxacyclobutene The heterocyclic group comprises, butyryl, dithioheterocyclic butadienyl, dioxolane, oxazolyl, thiazolyl, triazinyl, isothiazolyl, isoxazolyl, azahexyl, azahexyl, morpholinyl, oxadiazolyl, oxadiazolyl, oxazolyl, oxazolyl, oxazolyl, oxadiazinyl, oxazolyl, oxazolyl, oxazolyl, oxazolyl, oxazolyl, piperidinyl, piperidinyl, pyranyl, pyrazolinyl, pyrazolyl, pyrrolinyl, pyrrolyl, quininecycloyl, thiomorpholinyl, tetrahydropyranyl, tetrahydrofuranyl, tetrahydrothiophenyl, thiadiazolinyl, thiadiazolyl, thiadiazolyl, thiazolinyl, thiazolinyl, thiazolyl, thiomorpholinyl, 1,1-dioxothiomorpholinyl (thiomorpholinone), thiopyranyl, and trithiaylyl. The heterocyclic group may optionally be substituted by one or more substituents as described below.

[0060] Heterocyclic alkyl alkyl As used herein, the term "heterocyclic alkyl alkyl" refers to an alkyl group that is substituted by one or more heterocyclic alkyl (i.e., heterocyclic) groups.

[0061] hydroxyl The term "hydroxyl" is a technical term and, as used in this article, refers to -OH.

[0062] Hydroxyalkyl As used herein, the term "hydroxyalkyl" means at least one hydrocarbon group attached to a parent molecule portion by an alkyl group as defined herein. Representative examples of hydroxyalkyl groups include, but are not limited to, hydroxymethyl, 2-hydroxyethyl, 3-hydroxypropyl, 2,3-dihydroxypentyl, and 2-ethyl-4-hydroxyheptyl.

[0063] Improve , Increase , Inhibit , Decrease or reduce (reduce)As used herein, the terms “improve,” “increase,” “suppress,” “reduce,” and “lower,” and their grammatical equivalents, indicate differences in nature or quantity from the reference values.

[0064] Connector: As used herein, "linker" refers to the portion of a multi-element agent that connects different elements to one another. For example, those skilled in the art will understand that peptides whose structures comprise two or more functional or tissue domains typically include amino acid sequence segments between such domains that connect them to one another. In some embodiments, peptides including linker elements have an overall structure of the general formula S1-L-S2, wherein S1 and S2 may be the same or different and represent two domains associated with each other by the linker. In some embodiments, the length of the peptide linker is at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100 or more amino acids. In some implementations, the linker is characterized by its tendency to provide flexibility to the peptide rather than employing a rigid three-dimensional structure. A variety of different linker elements that can be suitably used in the engineering of peptides (e.g., fusion peptides) are known in the art (see, for example, Holliger, P. et al., (1993) Proc. Natl. Acad. Sci. USA 90:6444-6448; Poljak, RJ et al., (1994) Structure2: 1121-1123).

[0065] Nucleic acidAs used herein, the term "nucleic acid" in its broadest sense refers to any compound and / or substance that binds to or can bind to an oligonucleotide chain. In some embodiments, nucleic acids are compounds and / or substances that bind to or can bind to an oligonucleotide chain via phosphodiester bonds. As will become clear from the context, in some embodiments, the term nucleic acid refers to a single nucleic acid residue (e.g., a nucleotide and / or nucleoside), and in some embodiments, it refers to a polynucleotide chain comprising multiple individual nucleic acid residues. Nucleic acids can be or include DNA, RNA, or any combination thereof. Nucleic acids can comprise native nucleic acid residues, nucleic acid analogs, and / or synthetic residues. In some embodiments, nucleic acids comprise native nucleotides (e.g., adenosine, thymidine, guanosine, cytidine, uridine, deoxyadenosine, deoxythymidine, deoxyguanosine, and deoxycytidine). In some embodiments, the nucleic acid is or comprises one or more nucleotide analogs (e.g., 2-aminoadenosine, 2-thiothymidine, inosine, pyrrolopyrimidine, 3-methyladenosine, 5-methylcytidine, C-5-propynyl-cytidine, C-5-propynyl-uridine, 2-aminoadenosine, C5-bromouridine, C5-fluorouridine, C5-iodouridine, C5-propynyl-uridine, C5-propynyl-cytidine, C5-methylcytidine, 2-aminoadenosine, 7-deazoadenosine, 7-deazoguanosine, 8-oxoadenosine, 8-oxoguanosine, O(6)-methylguanine, 2-thiocytidine, methylated bases, intercalated bases, and combinations thereof). In some embodiments, the nucleic acid has a nucleotide sequence encoding a functional gene product (such as RNA or protein). In some embodiments, the nucleic acid comprises one or more introns. In some embodiments, the nucleic acid comprises one or more genes. In some embodiments, nucleic acids are prepared by one or more of the following: isolation from natural sources, enzymatic synthesis (in vivo or in vitro) via complementary template-based polymerization, replication in recombinant cells or systems, and chemical synthesis. In some embodiments, nucleic acid analogs differ from nucleic acids in that they do not utilize a phosphodiester backbone. For example, in some embodiments, nucleic acids may comprise one or more peptide nucleic acids that are known in the art and have peptide bonds instead of phosphodiester bonds in the backbone. Alternatively or additionally, in some embodiments, nucleic acids have one or more thiophosphate and / or 5'-N-phosphamide bonds instead of phosphodiester bonds. In some embodiments, nucleic acids contain one or more modified sugars (e.g., 2'-fluororibose, ribose, 2'-deoxyribose, arabinose, and hexose) compared to those sugars in natural nucleic acids.In some implementations, the nucleic acid is or contains at least 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 20, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 600, 700, 800, 900, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, 5000 or more residues. In some embodiments, the nucleic acid is partially or entirely single-stranded, or partially or entirely double-stranded. In some embodiments, the nucleic acid has a nucleotide sequence comprising at least one element that encodes a polypeptide or a complementary sequence to a sequence encoding a polypeptide. In some embodiments, the nucleic acid has enzymatic activity.

[0066] Pharmaceutically acceptable: As used herein, the term “pharmaceutically acceptable” applies to one or more or all of the components of the compositional formulation disclosed herein, meaning that each component must be compatible with the other ingredients in the composition and harmless to its recipient.

[0067] Pharmaceutically acceptable carriers: As used herein, the term “pharmaceuticalally acceptable carrier” refers to pharmaceutically acceptable materials, compositions, or media, such as liquid or solid fillers, diluents, excipients, or solvent encapsulating materials, that contribute to the formulation of a drug (e.g., a pharmaceutical preparation), alter the bioavailability of a drug, or facilitate the transport of a drug from one organ or part of a subject to another organ or part of the subject. Some examples of materials that can be used as pharmaceutically acceptable carriers include: sugars, such as lactose, glucose, and sucrose; starches, such as corn starch and potato starch; cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; astragalus gum powder; 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 non-toxic and compatible substances used in pharmaceutical formulations.

[0068] Pharmaceutical compositions or preparations As used herein, the terms “pharmaceutical composition” or “formulation” refer to a composition in which a therapeutic agent is formulated together with one or more pharmaceutically acceptable carriers.

[0069] phosphoryl group The term "phosphoryl group" is a technical term and, as used herein, can generally be represented by the following formula: , Where Q50 represents S or O, and R59 represents hydrogen, a lower alkyl group, or an aryl group; for example, -P(O)(OMe)- or -P(O)(OH)2. When used to replace, for example, alkyl groups, the phosphoryl group of a phosphoryl alkyl group can be represented by the following general formula: , Q50 and R59 are each defined independently as above, and Q51 represents O, S, or N; for example, -OP(O)(OH)OMe or -NH-P(O)(OH)2. When Q50 is S, the phosphoryl group is a "thiophosphate".

[0070] polypeptide: As used herein, a "peptide" refers to any polymer chain of amino acids. In some embodiments, the peptide has an amino acid sequence that is naturally occurring. In some embodiments, the peptide has an amino acid sequence that is not naturally occurring. In some embodiments, the peptide has an engineered amino acid sequence, i.e., it is designed and / or generated through artificial intervention. In some embodiments, the peptide may be or comprise natural amino acids, non-natural amino acids, or both. In some embodiments, the peptide may be or comprise only natural amino acids or only non-natural amino acids. In some embodiments, the peptide may comprise D-amino acids, L-amino acids, or both. In some embodiments, the peptide may comprise only L-amino acids. In some embodiments, the peptide may comprise one or more side groups or other modifications, such as one or more amino acid side chains, for example, at the N-terminus of the peptide, at the C-terminus of the peptide, at non-terminal amino acids, or any combination thereof. In some embodiments, such side groups or modifications may be selected from acetylation, amidation, esterification, methylation, phosphorylation, glycosylation, saccharification, sulfation, mannosylation, nitrosation, acylation, palmitoylation, isopentenylation, polyethylene glycolation, etc., including combinations thereof. In some embodiments, the polypeptide may be cyclic, and / or may contain a cyclic moiety.

[0071] In some embodiments, the term "peptide" may be attached to the name of a reference peptide, activity, or structure to indicate a peptide class sharing a common activity or structure. For such classes, this specification provides and / or those skilled in the art will understand that the amino acid sequence and / or function are exemplary peptides within the known class. In some embodiments, members of a peptide class or family exhibit significant sequence homology or identity with a reference peptide of that class, share common sequence motifs (e.g., characteristic sequence elements) with a reference peptide of that class, and / or share common activities (in some embodiments, at comparable levels or within a specified range) with a reference peptide of that class. For example, in some embodiments, the member polypeptide exhibits an overall degree of sequence homology or identity with the reference polypeptide of at least about 30%-40%, and typically greater than about 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or higher, and / or contains at least one region exhibiting very high sequence identity (typically greater than 90% or even 95%, 96%, 97%, 98%, or 99%) (e.g., in some embodiments, this may be or include a conserved region of a characteristic sequence element). Such a conserved region typically covers at least 3-4 amino acids, and in some cases up to 20 or more amino acids; in some embodiments, the conserved region covers at least one sequence segment of at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or more consecutive amino acids. In some embodiments, the related polypeptide may be or includes a fragment of the parent polypeptide. In some embodiments, the useful polypeptide may be or comprise multiple fragments, each of which is spatially arranged relative to each other in the same parent polypeptide in a different manner than it is spatially arranged in the polypeptide of interest (e.g., fragments that are directly linked in the parent polypeptide may be spatially separated in the polypeptide of interest, and vice versa, and / or the order in which the fragments are present in the polypeptide of interest may be different from that in the parent polypeptide), such that the polypeptide of interest is a derivative of its parent polypeptide.

[0072] refer to:As used herein, “reference” refers to a standard or control upon which comparisons are based. For example, in some embodiments, a drug, sample, sequence, subject, animal, or individual, or a group thereof, or a measure or characteristic representing them, is compared with a reference, drug, sample, sequence, subject, animal, or individual, or a group thereof, or a measure or characteristic representing them. In some embodiments, the reference is a measurement. In some embodiments, the reference is a defined standard or expected value. In some embodiments, the reference is a historical reference. The reference can be quantitative or qualitative. Generally, as understood by those skilled in the art, the reference and the value compared to it represent a measure under comparable conditions. Those skilled in the art will understand that when sufficient similarity exists, judgments and / or comparisons can be reasonably made based on that similarity. In some embodiments, an appropriate reference may be a drug, sample, sequence, subject, animal, or individual, or a group thereof, under conditions that those skilled in the art would consider comparable, for example, for the purpose of assessing one or more specific variables (e.g., the presence or absence of a drug or condition) or a measure or characteristic representing them.

[0073] sample: As used herein, the term "sample" generally refers to an aliquot of material obtained or derived from a source of interest. In some embodiments, the source of interest is of biological or environmental origin. In some embodiments, the sample is a "raw sample" obtained directly from the source of interest. In some embodiments, the term "sample" refers to a preparation obtained by treating a raw sample (e.g., by removing one or more components of the raw sample and / or by adding one or more agents to the raw sample). Such a "treated sample" may include, for example, cells, nucleic acids, or proteins extracted from a sample or obtained by subjecting the raw sample to techniques such as the separation and / or purification of certain components.

[0074] silane As used herein, the term "silyl" includes hydrocarbon derivatives of the silyl (H3Si-) group (i.e., (hydroyl)3Si–), where the hydrocarbon group is a monovalent group formed by removing a hydrogen atom from a hydrocarbon, such as ethyl or phenyl. The hydrocarbon group can be a combination of different groups, which can vary to provide a number of silyl groups, such as trimethylsilyl (TMS), tert-butyldiphenylsilyl (TBDPS), tert-butyldimethylsilyl (TBS / TBDMS), triisopropylsilyl (TIPS), and [2-(trimethylsilyl)ethoxy]methyl (SEM).

[0075] silyloxy As used herein, the term "silyloxy group" means, as defined herein, a silyl group attached to a parent molecule by an oxygen atom.

[0076] Specific binding: As used herein, the term "specific binding" refers to the ability to distinguish potential binding partners in the context in which binding will occur. A binder that interacts with a specific target in the presence of other potential targets is called a "specific binding agent." Specific binding "To the target that interacts with the binder. In some embodiments, specific binding is assessed by detecting or determining the degree of association between the binder and its partner; in some embodiments, specific binding is assessed by detecting or determining the degree of dissociation of the binder-partner complex; in some embodiments, specific binding is assessed by detecting or determining the ability of the binder to compete for alternative interactions between its partner and another entity. In some embodiments, specific binding is assessed by performing such detection or determination within a concentration range."

[0077] Subjects: As used herein, the term "subject" refers to an organism, typically a mammal (e.g., a human, rat, or mouse). In some embodiments, the subject has a disease, symptom, or condition. In some embodiments, the subject is susceptible to a disease, symptom, or condition. In some embodiments, the subject exhibits one or more symptoms or characteristics of a disease, symptom, or condition. In some embodiments, the subject does not have a disease, symptom, or condition. In some embodiments, the subject does not exhibit any symptoms or characteristics of a disease, symptom, or condition. In some embodiments, the subject has one or more characteristics that constitute susceptibility or risk to a disease, symptom, or condition. In some embodiments, the subject is a subject who has been tested for a disease, symptom, or condition and / or has been given a therapy. In some cases, a human subject may be referred to interchangeably as a "patient" or an "individual." A subject who receives a medication related to the treatment of a disease, symptom, or condition relevant to the subject may be referred to as a subject who requires the medication, i.e., a subject in need.

[0078] Replaced, or replaced by It should be understood that, as used herein with respect to molecular structure, “substitution” or “replaced by” includes the implicit condition that such substitution is in accordance with the permissible valence of the substituted atom and the substituent, and that the substitution produces a stable compound, for example, that is not spontaneously transformed by, for example, rearrangement, fragmentation, decomposition, cyclization, elimination or other reactions, except where cyclization is required under this disclosure.

[0079] ReplacementAs used herein, the term "substituted" in relation to molecular structure is also considered to include all permissible substituents in organic compounds. In a broad sense, permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents in organic compounds. Illustrative substituents include, for example, those described above. Permissible substituents may be one or more and may be the same or different for a suitable organic compound. For the purposes of this invention, heteroatoms such as nitrogen may have hydrogen substituents and / or any permissible substituents in organic compounds that satisfy the valence of heteroatoms as described herein. This invention is not intended to be limited in any way to the permissible substituents of organic compounds.

[0080] Therapeutic agents: As used herein, the term "therapeutic agent" means any agent that, when administered to a subject, elicits the desired pharmacological effect. In some embodiments, an agent is considered a therapeutic agent if it exhibits a statistically significant effect in an appropriate population. In some embodiments, an appropriate population may be a population of model organisms or a human population. In some embodiments, an appropriate population may be defined by various criteria, such as an age group, sex, genetic background, pre-existing clinical condition, etc. In some embodiments, a therapeutic agent is a substance that can be used to treat a disease, symptom, or illness. In some embodiments, a therapeutic agent is an agent that has been or requires approval by a government agency before it can be marketed for human administration. In some embodiments, a therapeutic agent is an agent that requires a medical prescription for human administration.

[0081] Treatment plan As used in this article, “treatment regimen” refers to a dosing regimen in the relevant population that can be associated with the desired or beneficial treatment outcome.

[0082] Effective therapeutic dose:As used herein, "therapeutic effective amount" refers to an amount that produces the desired effect of administration. In some embodiments, the term refers to an amount sufficient to treat a disease, symptom, and / or condition when administered to a population suffering from or susceptible to the disease, symptom, and / or condition according to a therapeutic dosing regimen. In some embodiments, a therapeutic effective amount is an amount that reduces the incidence and / or severity of one or more symptoms of a disease, symptom, and / or condition, and / or delays the onset of the disease, symptom, and / or condition. Those skilled in the art will understand that a therapeutic effective amount does not necessarily achieve successful treatment in every particular individual. Rather, a therapeutic effective amount can be an amount that provides a specific desired pharmacological response in a significant number of subjects when administered to a patient requiring such treatment. In some embodiments, reference to a therapeutic effective amount can refer to an amount measured in one or more specific tissues (e.g., tissues affected by the disease, symptom, or condition) or fluids (e.g., blood, saliva, serum, sweat, tears, urine, etc.). Those skilled in the art will understand that in some embodiments, a particular agent or therapy of a therapeutic effective amount may be formulated and / or administered in a single dose. In some implementations, the therapeutically effective agent may be formulated and / or administered in multiple doses, for example, as part of a dosing regimen.

[0083] thiocarbonyl As used in this article, the term "thiocarbonyl" refers to -C(=S)-.

[0084] treat As used herein, the term "treatment" (and treatment or treating) means the administration of a therapy that partially or completely reduces, improves, alleviates, or inhibits one or more symptoms, features, and / or causes of a particular disease, symptom, or condition; delays the onset of a particular disease, symptom, or condition; reduces the severity of a particular disease, symptom, or condition; and / or reduces the incidence of a particular disease, symptom, or condition, or is administered for the purpose of achieving any such outcome. In some embodiments, such treatment may be for subjects who do not exhibit signs of the relevant disease, symptom, or condition and / or only exhibit early signs of the disease, symptom, or condition. Alternatively or additionally, such treatment may be for subjects who exhibit one or more identified signs of the relevant disease, symptom, and / or condition. In some embodiments, treatment may be for subjects who have been diagnosed with the relevant disease, symptom, and / or condition. In some embodiments, treatment may be for subjects known to have one or more susceptibility factors that are statistically associated with an increased risk of developing the relevant disease, symptom, or condition. For the avoidance of doubt, the terms “disease,” “symptom,” and “illness,” and any equivalent thereof, are used interchangeably, such that, unless otherwise stated, the use of any one constitutes the use of its alternative form.

[0085] Cyclic peptides This disclosure includes the discovery of compositions comprising cyclic peptides that selectively target adipose tissue, as shown herein. This disclosure includes the understanding that, in various embodiments, the cyclic peptides provided herein can be used to deliver relevant pharmaceutical agents to adipose tissue, for example, for the treatment and / or diagnosis of diseases (e.g., diseases related to adipose tissue).

[0086] In various embodiments, the cyclic peptides of this disclosure are or comprise sequences selected from SEQ ID NO: 1-161 as shown herein (e.g., in Table 1), or variants thereof. In various embodiments, this disclosure provides a cyclic peptide that is or comprises a polypeptide differing from sequences selected from SEQ ID NO: 1-161 by one or more amino acid sequence differences. In various embodiments, an amino acid difference refers to a deletion (the removal of one amino acid), an insertion (the insertion of one amino acid), or a substitution (the replacement of one amino acid with another different amino acid). In various embodiments comprising multiple amino acid differences, each amino acid difference may be independently selected from, for example, deletion, insertion, or substitution. A sequence having one or more amino acid differences from sequences disclosed herein (e.g., reference sequences and / or sequences shown in Table 1) may be referred to, for example, as a “variant” or “mutant.”

[0087] In various embodiments, this disclosure provides a cyclic peptide that is or includes a polypeptide that differs from the sequence selected from SEQ ID NO: 1-161 by at least 1, at least 2, at least 3, at least 4, or at least 5 amino acid sequences. In various embodiments, this disclosure provides a cyclic peptide that is or includes a polypeptide that differs from the sequence selected from SEQ ID NO: 1-161 by exactly 1, exactly 2, exactly 3, exactly 4, or exactly 5 amino acid sequences. In various embodiments, this disclosure provides a cyclic peptide that is or includes a polypeptide that differs from the sequence selected from SEQ ID NO: 1-161 by 1 to 5 amino acid differences, 1 to 4 amino acid differences, 1 to 3 amino acid differences, or 1 to 2 amino acid differences.

[0088] In various embodiments, this disclosure provides a cyclic peptide that is or includes a polypeptide having a sequence selected from SEQ ID NO: 1-161, and / or includes a variant of a polypeptide having a sequence selected from SEQ ID NO: 1-161 (e.g., containing at least one amino acid sequence difference compared to a sequence selected from SEQ ID NO: 1-161), wherein the N-terminal amino acid of the cyclic peptide is a cysteine ​​residue, the C-terminal amino acid of the cyclic peptide is a cysteine ​​residue, and / or both the N-terminal and C-terminal amino acids of the cyclic peptide are cysteine ​​residues.

[0089] In various embodiments, the cyclic peptide of this disclosure is or comprises a sequence according to SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 as shown herein (e.g., in Table 1), or a variant thereof. In various embodiments, this disclosure provides a cyclic peptide that is or comprises a polypeptide that differs from SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 by one or more amino acid sequence differences.

[0090] In various embodiments, this disclosure provides a cyclic peptide that is or includes a polypeptide that differs from SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 by at least 1, at least 2, at least 3, at least 4, or at least 5 amino acid sequences. In various embodiments, this disclosure provides a cyclic peptide that is or includes a polypeptide that differs from SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 by exactly 1, exactly 2, exactly 3, exactly 4, or exactly 5 amino acid sequences. In various embodiments, this disclosure provides a cyclic peptide that is or includes a polypeptide that differs from SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 by 1 to 5 amino acid differences, 1 to 4 amino acid differences, 1 to 3 amino acid differences, or 1 to 2 amino acid differences.

[0091] In various embodiments, the cyclic peptide of this disclosure comprises the amino acid sequence according to SEQ ID NO: 1. In various embodiments, the cyclic peptide of this disclosure comprises an amino acid sequence differing from SEQ ID NO: 1 by no more than 1, 2, 3, 4, or 5 amino acids. In various embodiments, the cyclic peptide of this disclosure comprises an amino acid sequence having no more than 1, 2, 3, 4, or 5 amino acid substitutions, deletions, and / or insertions compared to the cyclic peptide of SEQ ID NO: 1.

[0092] In various embodiments, the cyclic peptide of this disclosure comprises the amino acid sequence according to SEQ ID NO: 2. In various embodiments, the cyclic peptide of this disclosure comprises an amino acid sequence differing from SEQ ID NO: 2 by no more than 1, 2, 3, 4, or 5 amino acids. In various embodiments, the cyclic peptide of this disclosure comprises an amino acid sequence having no more than 1, 2, 3, 4, or 5 amino acid substitutions, deletions, and / or insertions compared to the cyclic peptide of SEQ ID NO: 2.

[0093] In various embodiments, the cyclic peptide of this disclosure comprises the amino acid sequence according to SEQ ID NO: 3. In various embodiments, the cyclic peptide of this disclosure comprises an amino acid sequence differing from SEQ ID NO: 3 by no more than 1, 2, 3, 4, or 5 amino acids. In various embodiments, the cyclic peptide of this disclosure comprises an amino acid sequence having no more than 1, 2, 3, 4, or 5 amino acid substitutions, deletions, and / or insertions compared to the cyclic peptide of SEQ ID NO: 3.

[0094] In various embodiments, variants of the cyclic peptide sequences disclosed herein (e.g., sequences disclosed in Table 1 and / or SEQ ID NO: 1-161) target adipose tissue. In various embodiments, variants of the cyclic peptide sequences disclosed herein (e.g., sequences disclosed in Table 1 and / or SEQ ID NO: 1-161) target adipose tissue at a level equivalent to a reference sequence (e.g., a reference sequence selected from SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3). Adipose tissue targeting can be determined based on any binding assay or functional activity assay disclosed herein or known in the art. To provide a non-limiting example, adipose targeting can be measured and / or compared based on the delivery and / or function of exemplary inhibitory nucleic acids as shown herein.

[0095] Table 1. Selective cyclic peptide sequences in adipose tissue

[0096] The symbol # indicates a cysteine ​​residue that forms a disulfide bond. The two Cys residues marked with # form a disulfide bond.

[0097] For the avoidance of doubt, this disclosure includes peptide sequences larger than one or more of the cyclic peptides disclosed in SEQ ID NO: 1-161 and / or larger than one or more variants thereof. In various embodiments, the sequence of the cyclic peptide of this disclosure or a variant thereof may be present in a larger polypeptide that is partially or fully cyclic (e.g., the continuous backbone of the larger polypeptide is partially or fully cyclic). In various such embodiments in which a sequence selected from SEQ ID NO: 1-161 or a variant thereof is present in a larger polypeptide that is partially cyclic, a sequence selected from SEQ ID NO: 1-161 or a variant thereof is present in the cyclic portion of the polypeptide. It will be understood from this disclosure that a sequence selected from SEQ ID NO: 1-161 or a variant thereof may be referred to as “cyclic” even when present as part of a larger cyclic sequence. Therefore, this disclosure includes embodiments in which “terminal” amino acids of a sequence selected from SEQ ID NO: 1-161 or a variant thereof are directly linked to each other, and other embodiments in which additional amino acids of the larger cyclic polypeptide are positioned between such “terminal” amino acids as linkers.

[0098] Therefore, this disclosure contemplates and includes embodiments in which the cyclic peptide or the cyclic portion of a peptide of this disclosure may have a length of, for example, 3 to 40 or more amino acids. In some embodiments, the cyclic peptide of this disclosure may have a length of 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40 or more amino acids. In some embodiments, the cyclic peptide or the cyclic portion of a peptide of this disclosure may have a length of, for example, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16 amino acids. In some embodiments, the cyclic peptide or the cyclic portion of the peptide disclosed herein may have a length of, for example, about 7, about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15 or about 16 amino acids.

[0099] In some embodiments, the cyclic peptide or the cyclic portion of the peptide disclosed herein may have a length of 5 to 40 amino acids, 5 to 35 amino acids, 5 to 30 amino acids, 5 to 25 amino acids, 5 to 20 amino acids, 5 to 19 amino acids, 5 to 18 amino acids, 5 to 17 amino acids, 5 to 16 amino acids, 5 to 15 amino acids, 5 to 14 amino acids, 5 to 13 amino acids, 5 to 12 amino acids, 5 to 11 amino acids, 5 to 10 amino acids, 5 to 9 amino acids, or 5 to 8 amino acids. In some embodiments, the cyclic peptide or the cyclic portion of the peptide disclosed herein may have a length of 6 to 40 amino acids, 6 to 35 amino acids, 6 to 30 amino acids, 6 to 25 amino acids, 6 to 20 amino acids, 6 to 19 amino acids, 6 to 18 amino acids, 6 to 17 amino acids, 6 to 16 amino acids, 6 to 15 amino acids, 6 to 14 amino acids, 6 to 13 amino acids, 6 to 12 amino acids, 6 to 11 amino acids, 6 to 10 amino acids, 6 to 9 amino acids, or 6 to 8 amino acids. In some embodiments, the cyclic peptide or the cyclic portion of the peptide disclosed herein may have a length of 7 to 40 amino acids, 7 to 35 amino acids, 7 to 30 amino acids, 7 to 25 amino acids, 7 to 20 amino acids, 7 to 19 amino acids, 7 to 18 amino acids, 7 to 17 amino acids, 7 to 16 amino acids, 7 to 15 amino acids, 7 to 14 amino acids, 7 to 13 amino acids, 7 to 12 amino acids, 7 to 11 amino acids, 7 to 10 amino acids, 7 to 9 amino acids, or 7 to 8 amino acids. In some embodiments, the cyclic peptide or the cyclic portion of the peptide disclosed herein may have a length of 8 to 40 amino acids, 8 to 35 amino acids, 8 to 30 amino acids, 8 to 25 amino acids, 8 to 20 amino acids, 8 to 19 amino acids, 8 to 18 amino acids, 8 to 17 amino acids, 8 to 16 amino acids, 8 to 15 amino acids, 8 to 14 amino acids, 8 to 13 amino acids, 8 to 12 amino acids, 8 to 11 amino acids, 8 to 10 amino acids, 8 to 9 amino acids, or 8 to 8 amino acids.

[0100] In various embodiments, the preparation of cyclic peptides with 40 or more amino acids can be more challenging than the preparation of cyclic peptides with fewer than 40 amino acids. For cyclic peptides with fewer than 40 amino acids, such as those with 4 to 30 or 4 to 20 amino acids, certain advantageous properties of the cyclic peptides disclosed herein can also be more pronounced. In various embodiments, the cyclic peptides disclosed herein have a length of less than 40 amino acids, less than 30 amino acids, or less than 20 amino acids. In various embodiments, the cyclic peptides disclosed herein have a length of less than 15 amino acids. In various embodiments, the cyclic peptides disclosed herein have a length of 9 to 11 amino acids.

[0101] In various embodiments, the cyclic peptides of this disclosure have an amino acid number of 3 to 30 amino acids between two cysteine ​​residues (e.g., designated herein as two cysteine ​​residues forming a disulfide bond). In various embodiments, the cyclic peptides of this disclosure have an amino acid number equal to or about 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 amino acids between two cysteine ​​residues (e.g., designated herein as two cysteine ​​residues forming a disulfide bond). In various embodiments, the cyclic peptide of this disclosure has an amino acid number of 3 to 20, 3 to 15, 3 to 14, 3 to 13, 3 to 12, 3 to 11, 3 to 10, 3 to 9, 3 to 8, 3 to 7, 3 to 6, 3 to 5, and / or 3 to 4 amino acids between two cysteine ​​residues (e.g., designated herein as two cysteine ​​residues forming a disulfide bond). In various embodiments, the cyclic peptide of this disclosure has an amino acid number of 5 to 20, 5 to 15, 5 to 14, 5 to 13, 5 to 12, 5 to 11, 5 to 10, 5 to 9, 5 to 8, 5 to 7, 5 to 6, 5 to 5, and / or 5 to 4 amino acids between two cysteine ​​residues (e.g., designated herein as two cysteine ​​residues forming a disulfide bond). In various embodiments, the cyclic peptides of this disclosure have an amino acid number of 7 to 20, 7 to 15, 7 to 14, 7 to 13, 7 to 12, 7 to 11, 7 to 10, 7 to 9, 7 to 8, 7 to 7, 7 to 6, 7 to 5, and / or 7 to 4 amino acids between two cysteine ​​residues (e.g., designated herein as two cysteine ​​residues forming a disulfide bond). In various embodiments, the cyclic peptides of this disclosure contain 5 to 9 amino acids between the cysteine ​​residues of the cyclic peptides forming one or more disulfide bridges. In various embodiments, the cyclic peptides of this disclosure contain 7 amino acids between the cysteine ​​residues of the cyclic peptides forming one or more disulfide bridges.

[0102] Methods for peptide cyclization and obtaining cyclic peptides are known in the art. Cyclic peptides according to embodiments of the present invention can be prepared by procedures known to those skilled in the art. For example, cyclic peptides can be prepared using standard solid-phase peptide synthesis (SPPS) techniques, as described, for example, in Joo. (2012) Biomol Ther, 20(1):19-26, the contents of which are incorporated herein by reference. In various embodiments, the cyclic peptides disclosed herein can be peptides in which a bridge or link is formed between two amino acids that are part of or constitute a peptide. The bridge can be formed between amino acids having reactive groups (in addition to the amino and carboxyl groups required for the respective amino acids), for example, thiol groups. As understood by those skilled in the art, peptides containing two or more amino acids having such reactive groups can be cyclized. For example, a peptide containing two amino acids having thiol groups can be cyclized under conditions in which a disulfide bridge is formed between the thiol groups of the two amino acids containing thiol groups. Examples of amino acids having thiol groups and thus capable of forming bridges (i.e., disulfide bridges) include, but are not limited to, cysteine. In various embodiments, the cyclic peptides disclosed herein comprise two or more cysteine ​​residues. In various embodiments, the cyclic peptides disclosed herein may comprise 2, 3, 4, 5, 6, 7, or 8 cysteine ​​residues. In various embodiments, the cyclic peptides disclosed herein comprise one or more disulfide bridges formed between thiol groups of amino acids (e.g., cysteine). In various embodiments, the cyclic peptides disclosed herein may comprise one, two, three, or four disulfide bridges formed between thiol groups of amino acids (e.g., cysteine). As understood by those skilled in the art, the cyclic peptides disclosed herein can be cyclized using any bridging, linking, or method system known in the art or described herein. In various embodiments, the cyclic peptides disclosed herein are cyclized due to the presence of naturally occurring amino acids (e.g., cysteine). In various embodiments, the cyclic peptides disclosed herein are cyclized due to the presence of one or more non-naturally occurring amino acids. In various embodiments, the cyclic peptides disclosed herein can be cyclized using lactam bonds.

[0103] In various embodiments, the cyclic peptides or conjugates of this disclosure do not target and / or do not significantly target non-adipose tissues. In various embodiments, the cyclic peptides or conjugates of this disclosure do not target and / or do not significantly target muscle tissue, brain tissue, heart tissue, or kidney tissue. In various embodiments, the cyclic peptides or conjugates of this disclosure do not target and / or do not significantly target one or more of skeletal muscle tissue, cardiac muscle tissue, and / or smooth muscle tissue. In various embodiments, the cyclic peptides or conjugates of this disclosure target adipose tissue at a higher level than non-adipose tissues (e.g., non-adipose tissues disclosed herein). In various embodiments, the levels are about 10-fold, about 9-fold, about 8-fold, about 7-fold, about 6-fold, about 5-fold, about 4-fold, about 3-fold, about 2-fold, about 1.5-fold, about 1.4-fold, about 1.3-fold, about 1.2-fold, and about 1.1-fold. In various embodiments, the cyclic peptides or conjugates of this disclosure do not selectively target one or more of skeletal muscle tissue, smooth muscle tissue, and / or cardiac muscle tissue.

[0104] Conjugation of cyclic peptides with therapeutic and / or diagnostic agents This disclosure includes cyclic peptides that are covalently or non-covalently associated with one or more pharmaceutical agents (such as one or more therapeutic agents and / or diagnostic agents). As disclosed herein, cyclic peptides that are covalently or non-covalently associated with one or more pharmaceutical agents may be referred to, for example, as “conjugated” or its grammatical equivalents.

[0105] Those skilled in the art will understand that this disclosure includes the important finding that the cyclic peptides provided herein (e.g., cyclic peptides that are or comprise sequences selected from SEQ ID NO: 1-161 or variants of such sequences) can be used to target adipose tissue, and thus the cyclic peptides are characterized by certain general utilities, such as general utilities in delivering pharmaceutical agents to adipose tissue. Therefore, this finding regarding the disclosed cyclic peptides can be applied to the delivery of a variety of pharmaceutical agents to adipose tissue when those pharmaceutical agents are conjugated with the cyclic peptides. Furthermore, since techniques for conjugating pharmaceutical agents with peptides are well known in the art, it will be understood that those skilled in the art can readily select from a variety of pharmaceutical agents or classes of pharmaceutical agents to conjugate and / or deliver to adipose tissue. This disclosure provides a wide range of suitable media for delivering such pharmaceutical agents or classes of pharmaceutical agents to adipose tissue, which can be conjugated with cyclic peptides in any of a variety of suitable manners known in the art.

[0106] As disclosed herein and throughout, the cyclic peptides of this disclosure can be conjugated to, for example, up to five agents (each independently selected from therapeutic and / or diagnostic agents), such as 1, 2, 3, 4, or 5 agents (each independently selected from therapeutic and / or diagnostic agents). In various embodiments, the cyclic peptides of this disclosure can be conjugated to 1 or 2, 1 to 3, 1 to 4, or 1 to 5 agents (each independently selected from therapeutic and / or diagnostic agents). In some embodiments, whether in the conjugation reaction or in a group of conjugated cyclic peptides, the molar ratio of the agent to the cyclic peptide is 1:10 to 10:1, 1:5 to 5:1, 1:4 to 4:1, 1:3 to 3:1, 2:3 to 3:2, 2:1 to 1:2, about 4:1, about 3:1, about 1:3, about 2:1, about 1:2, about 3:2, about 2:3, or about 1:1.

[0107] As disclosed herein and throughout, an agent (e.g., ASO or other agents) may be conjugated to, for example, up to five cyclic peptides, each independently selected from the cyclic peptides disclosed herein (e.g., selected from SEQ ID NO: 1-161 and variants thereof). In various embodiments, the agent may be conjugated to, for example, one, two, three, four, or five cyclic peptides, each independently selected from the cyclic peptides disclosed herein (e.g., selected from SEQ ID NO: 1-161 and variants thereof). In various embodiments, the agent may be conjugated to one or two, one to three, one to four, or one to five cyclic peptides, each independently selected from the cyclic peptides disclosed herein (e.g., selected from SEQ ID NO: 1-161 and variants thereof). In various embodiments, the agent may be conjugated to two or three, two to four, or two to five cyclic peptides, each independently selected from the cyclic peptides disclosed herein (e.g., selected from SEQ ID NO: 1-161 and variants thereof). In various embodiments, the agent may be conjugated to 3 or 4, or 3 to 5 cyclic peptides, each independently selected from the cyclic peptides disclosed herein (e.g., selected from SEQ ID NO: 1-161 and variants thereof). In various embodiments, the agent may be conjugated to 4 to 5 cyclic peptides, each independently selected from the cyclic peptides disclosed herein (e.g., selected from SEQ ID NO: 1-161 and variants thereof). In some embodiments, the molar ratio of the agent to the cyclic peptide is 1:1 to 1:10, 1:1 to 1:5, or 1:2 to 1:5. In some embodiments, the molar ratio of the agent to the cyclic peptide is about 1:2. In some embodiments, the molar ratio of the agent to the cyclic peptide is about 1:3.

[0108] In some embodiments, the agent covalently associates with the cyclic peptide. In some embodiments, the agent covalently associates with functional groups on the side chain of the cyclic peptide. In some embodiments, the agent covalently associates with the N-terminus of the cyclic peptide. In some embodiments, the agent covalently associates with the C-terminus of the cyclic peptide.

[0109] In some embodiments, the agent covalently associates directly with the cyclic peptide. In some embodiments, the agent covalently associates indirectly with the cyclic peptide. In some embodiments, the agent covalently associates directly with functional groups on the side chain of the cyclic peptide. In some embodiments, the agent covalently associates indirectly with functional groups on the side chain of the cyclic peptide. In some embodiments, the agent covalently associates directly with the N-terminus of the cyclic peptide. In some embodiments, the agent covalently associates indirectly with the N-terminus of the cyclic peptide. In some embodiments, the agent covalently associates directly with the C-terminus of the cyclic peptide. In some embodiments, the agent covalently associates indirectly with the C-terminus of the cyclic peptide.

[0110] In some embodiments, the pharmaceutical agent is covalently associated with the cyclic peptide via a thioether bond, disulfide bond, oxime, thiazolidinyl ether, hydrazone, amide bond, azido bond, or maleimide bond. In some embodiments, the covalent association between the cyclic peptide and the pharmaceutical agent is formed via a Diels-Alder reaction. In some embodiments, the covalent association between the cyclic peptide and the pharmaceutical agent is formed via click chemistry.

[0111] In some embodiments, the agent is indirectly covalently associated with the cyclic peptide via a linker. In some embodiments, the linker comprises alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl (i.e., heterocyclic), aryl, heteroaryl, aralkyl, heteroaryl, carboxyl, ketone, aldehyde, alkylphosphoryl, alkenyl, heteroalkenyl, cycloalkenyl, cyclohexeneyl, alkynyl, sulfonyl, hydroxyl, cyano, azide, carbonyl, halogen (e.g., –F, –Cl, -Br or –I), haloalkyl, silyl, silyloxy, amine, ether, thioether, phosphine, aminophosphate, formamide, ester, imine ester, amidine, thioester, sulfonamide, carbamate, urea, guanidine, thiourea, disulfide, oxime, hydrazine, acylhydrazine, hydrazone, diazabond, triazole, triazoline, tetrazine, amino acid, or combinations thereof.

[0112] In some embodiments, the connector is a cuttable connector. In some embodiments, the connector is a self-degrading connector. In some embodiments, the connector contains p-aminobenzyl alcohol (PAB). In some embodiments, the connector is bifunctional. In some embodiments, the connector is trifunctional.

[0113] In some embodiments, the connector comprises a thioether bond, a disulfide bond, an oxime, a thiazoline, a hydrazone, an amide bond, an azide bond, or a maleimide bond.

[0114] In some implementations, the connector includes C1-C 30 Alkyl, C2-C 20 Alkyl, C3-C 12 Alkyl, C6-C 12 Alkyl or C6 alkyl. In some embodiments, shorter carbon links can be used to attach the agent, where proximity to the cyclic peptide does not pose a problem. In some embodiments, longer carbon links can be used, where the agent must be appropriately spaced from the cyclic peptide.

[0115] In some embodiments, the connector comprises an alkylamine. In some embodiments, the connector comprises an alkyl phosphate. In some embodiments, the connector comprises an alkylamide. In some embodiments, the connector comprises a C6 alkylamine. In some embodiments, the connector comprises an amino-C6 alkyl phosphate.

[0116] In some embodiments, the connector comprises a product of the Diels-Alder reaction, a product of a copper(I)-catalyzed 1,3-dipolar cycloaddition, or a product of a click chemistry reaction.

[0117] In some embodiments, the agent is an antisense oligonucleotide (ASO), and a linker containing a C6 alkylamine links the cyclic peptide to the 5' end of the ASO.

[0118] In some embodiments, peptide conjugates are prepared by solid-phase synthesis. In other embodiments, peptide conjugates are prepared using an automated synthesizer.

[0119] In some embodiments, the agent does not covalently associate with the cyclic peptide. In some embodiments, the cyclic peptide and the agent associate via electrostatic interactions (e.g., ionic or hydrogen bonds). In some embodiments, the cyclic peptide and the agent associate via hydrophobic interactions.

[0120] In some embodiments, the peptide conjugate comprises an admixture of a cyclic peptide and a pharmaceutical agent. Therefore, in some embodiments, the peptide conjugate is prepared by a method comprising admixture of the cyclic peptide with a pharmaceutical agent.

[0121] Therefore, this disclosure includes covalent and non-covalent conjugation of pharmaceutical agents with the cyclic peptides of this disclosure. Therapeutic and diagnostic agents of this disclosure may include, for example, but not limited to, nucleic acids, peptides, or chemical agents. In various embodiments, the therapeutic or diagnostic agents of this disclosure may diagnose, monitor, or modulate (e.g., upregulate or downregulate) the expression, production, and / or activity of one or more targets (e.g., target nucleic acids, genes, peptides, proteins, or compounds) provided in Table 2 in tissues such as those disclosed herein (e.g., adipose tissue) and / or diseases.

[0122] The therapeutic agents disclosed herein include, for example, therapeutic nucleic acids, peptides, and chemical agents. The nucleic acids disclosed herein include, for example, repressive nucleic acids and nucleic acids encoding expression products (e.g., transgenes). Examples of repressive nucleic acids include, but are not limited to, small interfering RNA (siRNA), microRNA (miRNA), and repressive antisense oligonucleotides (ASO). In some embodiments, the nucleic acids disclosed herein comprise an adenosine deaminase acting on an oligonucleotide that recruits RNA (ADAR). Examples of expression products include, but are not limited to, repressive nucleic acids and proteins. Examples of peptide agents include, for example, bioactive peptides, proteins, protein complexes, and peptide mimics. Examples of chemical agents include, but are not limited to, small molecules, such as small molecules having known biological activity. In various embodiments, the therapeutic agent may be, for example, a chemotherapeutic agent. In various embodiments, the therapeutic agent may be, for example, a radionuclide.

[0123] In some embodiments, the inhibitory nucleic acids of this disclosure inhibit or downregulate, or are able to inhibit or downregulate, gene expression (e.g., expression of genes associated with adipose tissue-related diseases or conditions, or expression of genes according to Table 2, or expression of genes encoding proteins according to Table 2). In some embodiments, the inhibitory nucleic acid of this disclosure inhibits or downregulates, or is capable of inhibiting or downregulating, gene expression (e.g., expression of genes associated with adipose tissue-related diseases or conditions, or expression of genes encoding proteins according to Table 2), by at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100%. In some embodiments, the repressive nucleic acids of this disclosure inhibit or downregulate, or are capable of inhibiting or downregulating, gene expression (e.g., expression of genes associated with adipose tissue-related diseases or conditions, or expression of genes encoding proteins according to Table 2), for example, at least about 40%. In some embodiments, the repressive nucleic acids of this disclosure inhibit or downregulate, or are capable of inhibiting or downregulating, gene expression (e.g., expression of genes associated with adipose tissue-related diseases or conditions, or expression of genes encoding proteins according to Table 2), for example, at least about 60%. In some embodiments, the inhibitory nucleic acids of this disclosure inhibit or downregulate, or are capable of inhibiting or downregulating, gene expression (e.g., expression of genes associated with adipose tissue-related diseases or conditions, or expression of genes encoding proteins according to Table 2), for example, by about 20% to about 40%, about 20% to about 60%, about 20% to about 80%, about 20% to about 100%, about 40% to about 60%, about 40% to about 80%, about 40% to about 100%, about 60% to about 80%, or about 60% to about 100%.In some embodiments, the inhibitory nucleic acid of this disclosure inhibits or downregulates, or is able to inhibit or downregulate, gene expression (e.g., expression of genes associated with adipose tissue-related diseases or conditions, or expression of genes encoding proteins according to Table 2) within a percentage range having a lower bound selected from about 20%, about 30%, about 40%, about 50%, or about 60%, and an upper bound selected from 100%, about 80%, about 90%, about 80%, about 70%, about 60%, about 50%, about 40%, or about 30%).

[0124] In some embodiments, the agent is an antisense oligonucleotide (ASO). In some embodiments, the ASO is covalently associated with a cyclic peptide via its 5' end. In some embodiments, the ASO is covalently associated with a cyclic peptide via its 3' end. In some embodiments, the ASO is covalently associated with a cyclic peptide via a heterocyclic base of the ASO. In various embodiments, the antisense oligonucleotide comprises phosphatidylmorpholinonucleotide (PMO) or peptide nucleic acid (PNA). In various embodiments, the antisense oligonucleotide hybridizes with a target nucleic acid (e.g., a target nucleic acid according to Table 2, or a target nucleic acid encoding a protein or RNA according to Table 2) and achieves regulation of gene expression activity or function (such as transcription, translation, or splicing). Regulation of gene expression can be achieved, for example, by target degradation or occupation-based repression. An example of regulation of RNA target function achieved by degradation is RNase H-based degradation of target RNA upon hybridization with a DNA-like antisense compound. In various embodiments, the antisense oligonucleotide may contain one or more modifications. In various embodiments, the antisense oligonucleotide may contain one or more modifications to reduce degradation. In various embodiments, the modification of the antisense oligonucleotide is a sugar modification. Sugar modifications include, but are not limited to, 2′-O-(2-methoxyethyl), 2′-fluorine, locked nucleic acids, and ethylene-bridged nucleic acids. In various embodiments, the sugar modification is 2′-O-(2-methoxyethyl). In various embodiments, the antisense oligonucleotide also contains a phosphate thioester nucleoside internucleotide bond.

[0125] In various embodiments, the agent is adeno-associated virus (AAV). In some embodiments, AAV is not covalently associated with the cyclic peptide. In some embodiments, AAV is covalently associated with the cyclic peptide. In some embodiments, AAV is directly covalently associated with the cyclic peptide. In some embodiments, the cyclic peptide of this disclosure is present in the protein of AAV (e.g., within the N-terminus or C-terminus of the sequence of the protein of AAV). In some embodiments, the cyclic peptide of this disclosure is present in the structural protein of AAV (e.g., the capsid protein of AAV) (e.g., within the N-terminus or C-terminus of the sequence of the structural protein of AAV). In some embodiments, the cyclic peptide of this disclosure is present in the capsid protein of AAV (selected from VP1, VP2, or VP3) (e.g., within the N-terminus or C-terminus of the sequence of the capsid protein of AAV). In some embodiments, the cyclic peptide of this disclosure is present in the hypervariable region of the AAV capsid. Without wishing to be bound by any particular scientific theory, the AAV capsid comprises 12 hypervariable regions exposed on the capsid surface. In some embodiments, AAV is indirectly covalently associated with the cyclic peptide via a linker. In some embodiments, AAV is covalently associated with the cyclic peptide via a functional group (e.g., a chemical stalk) of the AAV capsid. In some embodiments, AAV is covalently associated with the cyclic peptide via a lysine or arginine residue on the AAV capsid. In some embodiments, the AAV capsid contains a non-natural amino acid. The non-natural amino acid can act as a chemical stalk to facilitate covalent association with the cyclic peptide. Therefore, in some embodiments, AAV is covalently associated with the cyclic peptide via a non-natural amino acid. In some embodiments, the non-natural amino acid contains an azide. In some embodiments, AAV is covalently associated with the cyclic peptide via an azide of the non-natural amino acid. In some embodiments, the cyclic peptide contains an alkyne. In some embodiments, the covalent association between AAV and the cyclic peptide is formed via a click chemistry reaction. In some embodiments, the covalent association between AAV and the cyclic peptide is formed via a click chemistry reaction between an alkyne and an azide. In some embodiments, the covalent association between AAV and the cyclic peptide is formed via a click chemistry reaction between the alkyne of the cyclic peptide and the azide of the AAV capsid. In various embodiments, AAV can be used as a medium, for example, for delivering nucleic acids (e.g., nucleic acids provided in Table 2, or nucleic acids encoding proteins or RNA provided in Table 2). In various embodiments, AAV can be used as a medium, for example, for delivering transgenes (e.g., transgenes provided in Table 2, or transgenes encoding proteins or RNA provided in Table 2).

[0126] In various embodiments, the pharmaceutical agent is a lipid nanoparticle (LNP). The LNP may comprise one or more lipid components. The LNP can be used as a carrier for a therapeutic agent and may comprise synthetic ionizable or cationic lipids, phospholipids, cholesterol, and polyethylene glycol (PEG) lipids. In some embodiments, the LNP is not covalently associated with a cyclic peptide. In some embodiments, the LNP is covalently associated with a cyclic peptide (e.g., via association between the cyclic peptide and an LNP component such as a synthetic ionizable or cationic lipid, phospholipid, cholesterol, or PEG lipid). In some embodiments, the LNP is directly covalently associated with a cyclic peptide. In some embodiments, the LNP is indirectly covalently associated with a cyclic peptide via a linker. In some embodiments, the LNP is covalently associated with a cyclic peptide via a lipid component of the LNP. In some embodiments, the LNP is covalently associated with a cyclic peptide via a carboxylic acid component of the LNP. In some embodiments, the LNP is covalently associated with a cyclic peptide via an amine component of the cyclic peptide. In some embodiments, the LNP is covalently associated with a cyclic peptide via an amide bond. In some embodiments, the covalent association between the LNP and the cyclic peptide is formed via an amidation reaction. In some embodiments, the covalent association between the LNP and the cyclic peptide is formed by the amine of the cyclic peptide and the carboxylic acid of the LNP.

[0127] In various embodiments, the nucleic acid agent is a small non-coding RNA (e.g., non-coding RNAs according to Table 2). In various embodiments, the small non-coding RNA includes, but is not limited to, siRNA or miRNA. In various embodiments, siRNA includes any nucleic acid molecule capable of inhibiting or downregulating gene expression (e.g., the expression of genes according to Table 2, or genes encoding proteins according to Table 2) or viral replication (e.g., by mediating RNA interference or gene silencing in a sequence-specific manner). In various embodiments, miRNA includes any type of interfering RNA, including but not limited to endogenous miRNAs and artificial miRNAs. Endogenous miRNAs are small RNAs naturally present in the genome that are capable of regulating the productive utilization of mRNA. Artificial miRNAs include any type of RNA sequence other than endogenous miRNAs that are capable of regulating the productive utilization of mRNA.

[0128] In various embodiments, the nucleic acid agent is a transgene (e.g., a transgene according to Table 2, or a transgene encoding a protein or RNA according to Table 2). In various embodiments, the transgene comprises a nucleic acid (e.g., DNA or RNA) sequence encoding a protein or RNA (e.g., functional non-coding RNA). In some embodiments, the transgene comprises an open reading frame encoding a protein or RNA (e.g., functional non-coding RNA). The transgene can be isolated from an organism and introduced into different organisms of the same or different species to produce a transgene product (e.g., protein or RNA). As used herein, in the context of DNA encoding a protein, the term transgene may or may not include untranscribed flanking regions, such as RNA transcription initiation signals, polyadenylation addition sites, terminators, promoters, or enhancers.

[0129] In various embodiments, the therapeutic agent comprises a peptide (e.g., peptides provided in Table 2). In various embodiments, the peptide comprises a protein agent or a peptide mimic agent. In various embodiments, the peptide or peptide mimic agent of this disclosure has a defined length. The peptide or peptide mimic agent of this disclosure may have, but is not limited to, a length of up to 10, 20, 30, 40, 50, 100, 150, 200, 250, 300, 400, 500, 600, 700, 800, 900, 1000, or 2000 residues. In various embodiments, the peptide or peptide mimic agent of this disclosure has a length of at least 5, 10, 20, 30, 50, 100, 150, 200, 250, or 300 residues.

[0130] In various embodiments, the therapeutic agents include small molecule agents. In various embodiments, small molecule agents include DNA damaging agents, agents that inhibit DNA synthesis, microtubule and tubulin binding agents, antimetabolites, oxidative damage inducers, anti-angiogenic agents, endocrine therapies, anti-estrogens, immunomodulators (such as Toll-like receptor agonists or antagonists), histone deacetylase inhibitors, signal transduction inhibitors (such as kinase inhibitors), heat shock protein inhibitors, retinoids, growth factor receptor inhibitors, antimitotic compounds, anti-inflammatory agents, cell cycle regulators, transcription factor inhibitors, and apoptosis inducers, as well as any combination thereof.

[0131] In various implementations, the therapeutic agents include chemotherapeutic agents. Examples of chemotherapeutic agents include, but are not limited to, methotrexate, daunomycin, mitomycin, cisplatin (cisplatinum or cis-diaminodichloroplatin(II) (CCDP)), vincristine, epirubicin, fluorouracil, verapamil, cyclophosphamide, cytosine arabinoside, aminopterin, bleomycin, and mitomycin C. C) Democolcine, etoposide, mithramycin, chlorambucil, melphalan, daunorubicin, doxorubicin, tamoxifen, paclitaxel, vincristine, vinblastine, camptothecin, actinomycin D, cytarabine, cobrestatin, and their derivatives.

[0132] The diagnostic agents disclosed herein may include, for example, labeled tissues (e.g., adipose tissue) and / or agents that can be used to diagnose one or more medical conditions. In various embodiments, the agents include imaging agents. The diagnostic agents disclosed herein may include, for example, but not limited to, fluorescent labels, luminescent labels, enzymes, or detectable tags. Examples of imaging agents include, but are not limited to, radioactive isotopes (e.g., 3 H, 14 C 35 S, 125 I, 1311) Fluorescent labels (e.g., FITC, rhodamine, lanthanide phosphors), MRI contrast agents (e.g., gadolinium chelates (Gd)), luminescent labels (such as luminol); enzyme labels (e.g., horseradish peroxidase, β-galactosidase, luciferase, alkaline phosphatase, acetylcholinesterase), biotinylate groups (which can be detected by labeled avidin (e.g., streptavidin containing a fluorescent marker), or enzyme activity detectable by optical or calorimetric methods), and predetermined polypeptide epitopes recognized by secondary reporter molecules (e.g., leucine zipper pairs, binding sites of secondary antibodies, metal-binding domains, epitope tags). In various embodiments, the imaging agent comprises any suitable radionuclide, including but not limited to... 227 Ac、 211 At、 131 Ba、 77 Br、 109 Cd, 51 Cr 67 Cu、 165 Dy、 155 Eu、 153 Gd, 198 Au、 166 Ho、 113m In、 115m In、 123 I, 125 I, 131 I, 189 Ir、 191 Ir、 192 Ir、 194 Ir、 52 Fe、 55 Fe、 59 Fe、 177 Lu、 109 Pd, 32 P, 226 Ra、 186 Re、 188 Re、 153 Sm、 46 Sc、 47 Sc、 72 Se、 75 Se、 105 Ag、 89 Sr、 35 S, 177 Ta、 117 mSn, 121 Sn、 166 Yb、 169 Yb、 90 Y、 212 Bi、 119 Sb,197 Hg, 100 Pd, 101m Rh and 212 Pb. In various embodiments, radionuclides can also be used to deliver therapeutic doses of radiation to tissues or cells.

[0133] Preparation and application of cyclic peptides and cyclic peptide conjugates The cyclic peptides and peptide conjugates disclosed herein can be used in a variety of applications (e.g., therapeutic or diagnostic applications) including, but not limited to, the treatment of diseases (e.g., diseases related to adipose tissue). This disclosure includes the broad understanding that the cyclic peptides and peptide conjugates of this disclosure can be used to deliver pharmaceutical agents to adipose tissue and treat diseases (e.g., diseases related to adipose tissue). Those skilled in the art will further understand from this disclosure that the cyclic peptides and peptide conjugates of this disclosure are not limited to the treatment of diseases related to adipose tissue, but can be used to treat any condition in any tissue to which the cyclic peptide has selectivity.

[0134] This disclosure includes pharmaceutical compositions for delivering one or more pharmaceutical agents to a subject. As disclosed herein, the pharmaceutical compositions may be in any form known in the art, including formulations for administration via any route known in the art. A suitable route of administration may be selected based on the age and condition of the subject.

[0135] The pharmaceutical compositions disclosed herein may be in the form of, for example, liquids, semi-solids, and solid dosage forms. The pharmaceutical compositions disclosed herein may be in the form of, for example, liquid solutions (e.g., solutions for injection and infusion), dispersions or suspensions, tablets, pills, powders, liposomes, and suppositories. The choice or use of any particular form may depend in part on the intended route of administration and therapeutic application. Thus, the compositions may be formulated for administration via parenteral route (e.g., intravenous, subcutaneous, intraperitoneal, or intramuscular injection). As used herein, parenteral administration refers to a route of administration other than enteral and local administration, typically by injection, and includes, but is not limited to, intravenous, intranasal, intraocular, intramuscular, intra-articular, intrathecal, intracapsular, intra-sacral, intraorbital, intracardiac, intradermal, intrapulmonary, intraperitoneal, tracheal, subcutaneous, subepidermal, intra-articular, subcapsular, subarachnoid, intraspinal, epidural, intracerebral, intracranial, carotid, and intrasternal injections and infusions.

[0136] In some embodiments, the compositions provided herein are available in unit dosage forms suitable for self-administration. Such unit dosage forms may be provided in containers (e.g., pills, vials, cartridges, pre-filled syringes, or disposable pens).

[0137] The pharmaceutical compositions disclosed herein may be in the form of injection or infusion. For example, this disclosure includes sterile formulations for injection, which may be formulated according to conventional pharmaceutical practices. Sterile solutions for injection may be prepared by incorporating the desired amount of the composition described herein into a suitable solvent having one or more of the ingredients listed above, followed by filtration sterilization as needed. Solutions for injection may be formulated as aqueous solutions for injection, for example using distilled water, physiological saline, or an isotonic solution containing glucose and other supplements (such as D-sorbitol, D-mannose, D-mannitol, or sodium chloride), optionally in combination with suitable solubilizers (e.g., alcohols (such as ethanol) and / or polyols (such as propylene glycol or polyethylene glycol) and / or nonionic surfactants (such as polysorbate 80™ or HCO-50, etc.). Regarding the sterile powder used to prepare the sterile solutions for injection, preparation methods include vacuum drying and freeze-drying, which produce powders of the compositions described herein, as well as any additional desired components from a previously sterile filtered solution of that powder (see below). Appropriate fluidity of the solution can be maintained, for example, by using coating (such as lecithin), or, in the case of a dispersion, by maintaining the desired particle size, and by using a surfactant. Prolonged absorption of the injectable composition can be achieved by including a delayed-absorption agent (e.g., monostearate and gelatin) in the composition. In certain cases, the pharmaceutical composition can be formulated into a buffer solution, for example, of suitable concentration and suitable for storage, for example, at 2°C–8°C (e.g., 4°C).

[0138] In various embodiments, the pharmaceutical compositions of this disclosure can be formulated as solutions, microemulsions, dispersions, liposomes, or other ordered structures suitable for stable storage at high concentrations. Typically, dispersions are prepared by incorporating the compositions described herein into a sterile medium containing a basic dispersion medium.

[0139] In various cases, pharmaceutical compositions may be formulated to include pharmaceutically acceptable carriers or excipients. Examples of pharmaceutically acceptable carriers include, but are not limited to, any and all physiologically compatible solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic agents, and absorption delay agents. Pharmaceutical compositions of therapeutic agents disclosed herein may include pharmaceutically acceptable salts (e.g., acid addition salts or base addition salts).

[0140] In some embodiments, the composition may be formulated with a carrier that protects the compound from rapid release, such as a controlled-release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid may be used. Many methods for preparing such formulations are known in the art. See, for example, JR Robinson (1978) “Sustained and Controlled Release Drug Delivery Systems,” Marcel Dekker, Inc., New York.

[0141] The route of administration can be parenteral, such as by injection, nasal administration, pulmonary administration, or percutaneous administration. Administration can be by intravenous injection, intramuscular injection, intraperitoneal injection, or subcutaneous injection. Administration can be systemic or local. In some embodiments, the compositions described herein can be therapeutically delivered to a subject via local administration. As used herein, “local administration” or “local delivery” can refer to delivery that does not rely on the transport of the composition or agent to the intended target tissue (e.g., adipose tissue) or site via the vascular system. For example, the composition can be delivered by injection or implantation of the composition or agent or by injection or implantation of a device containing the composition or agent. In some embodiments, after local application near the target tissue or site (e.g., adipose tissue), the composition or agent, or one or more components thereof, can diffuse to the intended target tissue or site outside the application site.

[0142] Pharmaceutical compositions can be administered parenterally in the form of injectable formulations comprising a sterile solution or suspension in water or another pharmaceutically acceptable liquid. For example, a pharmaceutical composition can be formulated by appropriately combining a therapeutic molecule with a pharmaceutically acceptable medium or medium (such as sterile water and saline, vegetable oil, emulsifier, suspension, surfactant, stabilizer, flavoring excipient, diluent, carrier, preservative, binder) and then mixing them in a unit dose form required by generally accepted pharmaceutical practices. Examples of oily liquids include sesame oil and soybean oil, and oily liquids can be combined with benzyl benzoate or benzyl alcohol as a solubilizer. Other items that may be included are buffers (such as phosphate buffers or sodium acetate buffers), soothing agents (such as procaine hydrochloride), stabilizers (such as benzyl alcohol or phenol), and antioxidants. The formulated injectable can be packaged in suitable ampoules.

[0143] In various embodiments, subcutaneous administration can be accomplished by means of a device such as a syringe, a pre-filled syringe, an autoinjector (e.g., a disposable or reusable autoinjector), a pen syringe, a patch syringe, a wearable syringe, a portable syringe infusion pump with a subcutaneous infusion element, or other devices for use in combination with a therapeutic agent administered subcutaneously.

[0144] The injection system disclosed herein can employ a delivery pen as described in U.S. Patent No. 5,308,341. Pen-type devices (most commonly used for self-delivery of insulin to diabetic patients) are well known in the art. Such devices may include at least one injection needle (e.g., a 31-gauge needle of about 5 to 8 mm in length), typically pre-filled with a therapeutic solution containing one or more therapeutic units, and may be used to rapidly deliver the solution to the subject with minimal pain. A drug delivery pen includes a vial holder in which a vial of therapeutic or other medication may be received. The pen may be a fully mechanical device, or it may be combined with electronic circuitry to accurately set and / or indicate the dosage of medication injected into the user. See, for example, U.S. Patent No. 6,192,891. In some embodiments, the needle of the pen-type device is disposable, and the cartridge includes one or more disposable replacement needles. Pen-type devices suitable for delivering compositions of any of the features of this invention are also described, for example, in U.S. Patent Nos. 6,277,099, 6,200,296, and 6,146,361, the disclosures of each of which are incorporated herein by reference in their entirety. Microneedle-based pen-type devices are described, for example, in U.S. Patent No. 7,556,615, the disclosure of which is incorporated herein by reference in its entirety. See also the MOLLY™ precision pen injector (PPI) device manufactured by Scandinavian Health Ltd.

[0145] In some embodiments, the compositions can be formulated to be suitable for intrapulmonary administration (e.g., for administration via inhaler or nebulizer) to mammals (such as humans). Methods for formulating such compositions are well known in the art. Dry powder inhaler formulations and suitable systems for administering the formulations are also known in the art. Lung administration can be oral and / or nasal. Examples of drug delivery devices for the lungs include metered-dose inhalers, dry powder inhalers (DPIs), and nebulizers. For example, the compositions described herein can be administered to the lungs of a subject via a dry powder inhaler. These inhalers are propellant-free devices that deliver a dispersible and stable dry powder formulation to the lungs. DPI devices have been used for the pulmonary administration of peptides such as insulin and growth hormone. In some embodiments, the compositions described herein can be administered intrapulmonaryly via a metered-dose inhaler. These inhalers rely on a propellant to deliver discrete doses of the compound to the lungs.

[0146] In some embodiments, the composition may be formulated and delivered to the eye, for example, in a pharmaceutically acceptable solution, suspension, or ointment. Preparations for treating the eye may be in the form of a sterile aqueous solution containing, for example, additional ingredients such as, but not limited to, preservatives, buffers, tensioning agents, antioxidants and stabilizers, nonionic wetting or clarifying agents, and thickeners. Preparations as described herein may be applied topically to the eye of a subject requiring treatment (e.g., a subject with AMD) by conventional methods, for example, in the form of drops, or by immersion of the eye in a therapeutic solution containing one or more of the compositions.

[0147] In some embodiments, various devices for delivering medication into the vitreous cavity of the eye can be adapted to administer compositions as described herein. For example, U.S. Publication No. 2002 / 0026176 describes a drug-filled plug that can be inserted through the sclera to protrude into the vitreous cavity for delivery of a drug into the vitreous cavity. In another example, U.S. Patent No. 5,443,505 describes an implantable device for introducing into the suprachoroidal space or avascular region for sustained release of a drug into the intraocular space.

[0148] In some embodiments, the compositions described herein can be applied topically to a joint (e.g., a hinged joint). For example, in embodiments where the symptom is arthritis, the therapeutically suitable composition can be applied directly to the joint (e.g., into the joint cavity) or near the joint. Examples of intra-articular joints to which the compositions described herein can be applied topically include, for example, the hip, knee, elbow, wrist, sternoclavicular, temporomandibular, carpal, tarsal, and ankle joints, as well as any other joints with arthritis. The compositions described herein can also be applied to bursae, such as the acromial bursa, biceps brachii radial bursa, elbow radial bursa, deltoid bursa, subpatellar bursa, ischial bursa, and any other bursa known in the medical field.

[0149] In some embodiments, the composition may be formulated for storage at temperatures below 0°C (e.g., -20°C or -80°C). In some embodiments, the composition may be formulated for storage at 2°C–8°C (e.g., 4°C) for up to 2 years (e.g., one month, two months, three months, four months, five months, six months, seven months, eight months, nine months, ten months, eleven months, one year, eleven and a half years, or two years). Therefore, in some embodiments, the composition described herein is stably stored at 2°C–8°C (e.g., 4°C) for at least one year.

[0150] Pharmaceutical compositions may comprise a therapeutically effective amount of the therapeutic agent described herein. This effective amount can be readily determined by those skilled in the art. A therapeutically effective amount may be an amount in which the beneficial therapeutic effect outweighs any toxic or adverse effects of the composition. In some embodiments, the dosage may also be selected to reduce or avoid the production of antibodies or other host immune responses against the therapeutic agent. Those skilled in the art will understand that data obtained from cell culture assays and animal studies can be used to formulate dosage ranges for human use. In various embodiments, the amount of active ingredient contained in the pharmaceutical preparation allows for the administration of an appropriate dose within a specified range to a subject. The dosage and method of administration may vary depending on the patient's weight, age, condition, and other characteristics, and may be appropriately selected by those skilled in the art as needed.

[0151] Pharmaceutical compositions comprising certain therapeutic agents (e.g., cyclic peptide conjugates as disclosed herein) may be administered at a fixed dose or at a dose of milligrams per kilogram (mg / kg). While not intended to be limiting, exemplary single doses of certain pharmaceutical compositions described herein may include amounts equal to, for example, 0.001 to 1000 mg / kg body weight, 1-1000 mg / kg body weight, 1-100 mg / kg body weight, 0.5-50 mg / kg body weight, 0.1-100 mg / kg body weight, 0.5-25 mg / kg body weight, 1-20 mg / kg body weight, and 1-10 mg / kg body weight of certain therapeutic agents as described herein. Exemplary doses of the compositions described herein include, but are not limited to, 0.1 mg / kg, 0.5 mg / kg, 1.0 mg / kg, 2.0 mg / kg, 4 mg / kg, 8 mg / kg, or 20 mg / kg. This disclosure is not limited to such ranges or doses.

[0152] Adipose tissue and its diseases As used in this article, adipose tissue refers to the tissue and cells characterized by the presence of adipocytes. Adipose tissue contributes to biological functions, including energy storage, energy homeostasis, and endocrine function, and may also contribute to certain diseases.

[0153] Examples of adipose tissue include white adipose tissue, brown adipose tissue, visceral adipose tissue, and subcutaneous adipose tissue. In various embodiments, white adipose tissue is characterized by the presence of white adipocytes. In various embodiments, white adipocytes are characterized by the presence of individual lipid droplets of triglycerides within the adipocytes. In various embodiments, white adipose tissue is distributed throughout the subject's body and includes visceral white adipose tissue and subcutaneous white adipose tissue. In various embodiments, visceral white adipose tissue is distributed around organs and provides a protective cushion. In various embodiments, subcutaneous white adipose tissue is located under the skin and provides thermal or cold insulation. In various embodiments, brown adipose tissue is characterized by the presence of brown adipocytes. In various embodiments, brown adipocytes are characterized by the presence of triglycerides in small and multiple vacuoles within the adipocytes. In various embodiments, brown adipose tissue is distributed around the interscapular region (e.g., in the upper back region), axillary region (e.g., below the shoulder joint), paravertebral region, and perirenal region. In various embodiments, adipose tissue may include adipocytes, preadipocytes, macrophages, neutrophils, lymphocytes, blood cells, and endothelial cells. In some embodiments, adipocytes specifically include white adipocytes and brown adipocytes. In some embodiments, adipose tissue specifically includes white adipose tissue.

[0154] Impaired adipose tissue function is associated with a variety of diseases. Delivery of pharmaceutical agents to adipose tissue can be used to treat such diseases. This disclosure specifically includes the use of the cyclic peptides of this disclosure for the delivery of pharmaceutical agents to adipose tissue (e.g., wherein the cyclic peptides are covalently or non-covalently conjugated to the pharmaceutical agent). Delivery of therapeutic or diagnostic agents to tissue can be used, for example, to treat or diagnose diseases associated with adipose tissue. Examples of diseases that can be used to treat or diagnose diseases associated with adipose tissue include, for example, obesity, cachexia, hyperglycemia, insulin resistance, type 2 diabetes, hypertension, cancer, heart disease, immune diseases, arthritis, diseases of the central nervous system, metabolic disorders, and age-related diseases.

[0155] In various embodiments, the disease or symptom can be treated by administration of the compositions, peptide conjugates, or agents targeting nucleic acids or proteins (e.g., disease-related nucleic acids (e.g., genes encoding proteins or RNA), disease-related proteins, or nucleic acids encoding such proteins) (e.g., agents covalently or non-covalently associated with the cyclic peptides of the present disclosure and / or therapeutic agents). In various embodiments, the target of the composition, peptide conjugate, or agent may be a target according to Table 2. In various embodiments, the target nucleic acid (e.g., a gene encoding a protein or RNA) may be a nucleic acid according to Table 2. In various embodiments, the target protein may be, or a protein encoded by the target nucleic acid may be a protein according to Table 2. In various embodiments, the target nucleic acid (e.g., a gene encoding a protein or RNA), the target protein, or the protein encoded by the target nucleic acid may be a nucleic acid or protein according to Table 2, and the compositions, peptide conjugates, or agents of the present disclosure may be used to treat the corresponding diseases indicated in Table 2. In some embodiments, the disease may be obesity, metabolic disorder, and / or cachexia.

[0156] In various embodiments, the diseases of this disclosure (e.g., diseases related to adipose tissue or impairment of adipose tissue function) are associated with the expression, production, and / or activity of one or more targets (e.g., target nucleic acids, genes, peptides, proteins, or compounds) according to Table 2, and / or can be treated by reducing the expression, production, and / or activity of these targets. In various embodiments, the expression, production, and / or activity of one or more targets in Table 2 are diagnosed, monitored, or modulated (e.g., upregulated or downregulated) by delivery of therapeutic or diagnostic agents disclosed herein.

[0157] Table 2. List of targets for adipose tissue-related diseases

[0158] Example Embodiments of the present invention demonstrate that cyclic peptides as shown in this disclosure can be used to deliver therapeutic agents to adipose tissue, wherein exemplary antisense oligonucleotides (ASOs) and small interfering RNA (siRNA) payloads of the present invention are representative. Embodiments of the present invention provide data demonstrating that the cyclic peptides of the present disclosure can effectively deliver therapeutic agents to adipose tissue, as illustrated by the delivery of representative payloads such as ASOs or siRNAs that regulate gene expression in adipose tissue cells. It will be understood from the embodiments of the present invention that those skilled in the art can select payloads that will provide therapeutically beneficial effects in the treatment of a disease.

[0159] Example 1: Delivery of a representative antisense oligonucleotide (ASO) payload to adipose tissue using the cyclic peptides of this disclosure. Embodiments of the present invention demonstrate that the cyclic peptides of this disclosure, conjugated to a representative ASO payload, can deliver the payload to target adipose tissue cells and modulate biological activity, demonstrating efficacy for treating diseases. Specifically, embodiments of the present invention demonstrate that using the cyclic peptides of this disclosure to deliver ASO to target adipose tissue cells achieves downregulation of target gene expression.

[0160] This disclosure utilizes the cyclic peptide (SEQ ID NO: 162) conjugated to an ASO targeting Malat1. Malat1 is a long non-coding RNA transcript whose upregulation is associated with the progression and development of a broad range of indications, including cancer, diabetes, and inflammatory diseases (Biswas, S. et al. (2018) Sci Rep, 8: 6526.). As indicated in Table 3, the Malat1 ASO contains phosphate thioester substitution and 2' sugar modification to inhibit nuclease degradation and promote mediated delivery to cells, and may contain one or more additional modifications.

[0161] The conjugation between exemplary cyclic peptides (SEQ ID NOs: 1-3 as indicated in Table 1) and exemplary antisense oligonucleotides (SEQ ID NO: 162 as indicated in Table 3) was performed using an amino C6 linker comprising an amino group and a six-carbon spacer. The C-terminus of the cyclic peptide was functionalized with an azide, and the 5' terminus of the ASO was functionalized with dibenzocyclooctyl (DBCO). Following copper-free click chemistry, peptide oligonucleotide conjugates were constructed at a cyclic peptide:ASO ratio of 1:1.

[0162] The conjugated cyclic peptides of the present invention, pre-reconstituted in isotonic saline, along with a non-targeted control and a PBS injection control, were delivered in vivo via tail vein injection to male BL6 mice aged 8–10 weeks. Mice received a total of eight injections of 1.5 mg / kg over 24 days. On day 27, three days after the last dose, mice were perfused to collect tissues (including adipose tissue, muscle tissue, brain tissue, heart tissue, and kidney tissue) for RNA or histological processing. For each collected tissue, tissue samples were rapidly frozen and homogenized at maximum speed for 3 minutes with SPEX SamplePrep and steel balls in the presence of BME. White adipose tissue (WAT) was processed using the Qiagen QIAzol kit, brain and kidney tissues using the Qiagen RNeasy kit, and muscle and heart tissues using the Qiagen RNeasy fibrous tissue kit.

[0163] Total RNA aliquots were prepared using Applied Biosystems' high-affinity cDNA reverse transcription mixture. The cDNA samples were then processed for qPCR using Applied Biosystems Taqman reagents. Target ASO gene (MALAT1) and housekeeping gene (GAPDH) were measured based on the drug administration in each mouse group. Applied Biosystems Taqman primers and probes IDs Mm01227912_s1 (MALAT1) and Mm99999915_g1 (GAPDH) were used. All treated tissues were analyzed. Cq analysis. When relevant, the target gene mRNA expression profile is visualized relative to the PBS control group and the non-targeted ASO control group.

[0164] Table 3. Exemplary antisense oligonucleotide sequences

[0165] This indicates the position of the thiophosphate bond. M indicates 2'-O-methylation (adding a methyl group to the 2' hydroxyl group of the ribose portion of the nucleoside). For example, MG indicates 2'-O-methylated guanosine.

[0166] like Figure 1 As shown, compared with the PBS control and the non-targeted control (where Malat1 ASO is conjugated with the control peptide), delivery of Malat1 ASO conjugated with the exemplary targeting cyclic peptide significantly reduced Malat1 expression in adipose tissue. When normalized to the Malat1 expression level of the non-targeted control in the tissue, Figure 2The results demonstrated that conjugation to the exemplary cyclic peptides resulted in an increase in the downregulation of Malat1 in adipose tissue, but not in muscle, brain, heart, or kidney tissue. In summary, these results indicate that the cyclic peptides of this disclosure can be used to deliver therapeutic agents to adipose tissue. The results show that the cyclic peptides disclosed herein, conjugated to a representative ASO payload targeting Malat1, can selectively target adipose tissue to knock down Malat1 expression. Therefore, the targeted cyclic peptides disclosed herein can be used to deliver payloads to target adipose tissue cells and modulate biological activity, demonstrating utility for treating diseases.

[0167] Example 2: Delivery of a representative small interfering RNA (siRNA) payload to adipose tissue using the cyclic peptide of this disclosure. Embodiments of the present invention demonstrate that the cyclic peptides of this disclosure, conjugated to a representative siRNA payload, can deliver the payload to target adipose tissue cells and modulate biological activity, demonstrating efficacy for treating diseases. Specifically, embodiments of the present invention demonstrate that using the cyclic peptides of this disclosure to deliver siRNA to target adipose tissue cells achieves downregulation of target gene expression.

[0168] With target ALDH2 The siRNA-conjugated peptides of this disclosure were tested as described herein. ALDH2 is a mitochondrial enzyme expressed in most human tissues, including adipose tissue, and is present at high levels in liver, heart, kidney, and muscle tissues (Mark J. et al. (1998) BBA, 1399: 181-186.). It belongs to the aldehyde dehydrogenase family of enzymes, and its activity is associated with a variety of indications, including cardiovascular disease, diabetes, neurodegenerative diseases, stroke, and cancer (Chen, CH et al. (2014) Physiol Rev, 94: 1-34.).

[0169] The conjugation between the exemplary cyclic peptide (SEQ ID NO: 3 as indicated in Table 1) and the exemplary siRNA was performed using an amino C6 linker comprising an amino group and a six-carbon spacer, as described in Example 1. The C-terminus of the cyclic peptide was functionalized with an azide, and the 5' terminus of the siRNA was functionalized with a dibenzocyclooctyl (DBCO). Following copper-free click chemistry, peptide-siRNA conjugates were constructed at a cyclic peptide:siRNA ratio of 1:1.

[0170] The conjugated cyclic peptide of the present invention, along with a PBS-injected control, was delivered subcutaneously to female C57BL / 6 (BL6) mice. Mice received a single dose of the conjugated cyclic peptide at 5 mg / kg, or PBS as a control. On day 7 post-injection, mice were perfused to collect tissues (including kidney, spleen, heart, muscle, and adipose tissue) for RNA processing. Muscle tissue included tibialis anterior (TA), triceps, and quadriceps muscles. Adipose tissue included gonadal white adipose tissue (gWAT) and subcutaneous white adipose tissue (scWAT). RNA processing was performed as described in Example 1. The mRNA expression of the target gene in the treated mice was normalized to the expression of the same gene in the PBS-treated control mice.

[0171] like Figure 3 As shown, compared to the PBS control, delivery of siRNA conjugated to the exemplary targeting cyclic peptide significantly reduced target gene expression in adipose tissue (gWAT and scWAT tissues). In mice treated with the conjugate, the percentage reduction in target gene expression in adipose tissue was greater than in other tissues evaluated.

[0172] These results indicate that the cyclic peptides of this disclosure can be used to deliver various types of therapeutic agents to adipose tissue. The results show that the cyclic peptides disclosed herein, conjugated to representative siRNA payloads, can selectively target adipose tissue to knock down expression. Therefore, the data demonstrate that the targeting cyclic peptides disclosed herein can be used to deliver payloads to target adipose tissue cells and modulate biological activity, demonstrating their utility for treating diseases.

[0173] Other implementation plans It should be understood that the scope of this disclosure is defined by what should be understood from this disclosure and the claims, and not by the specific embodiments presented by way of examples. Elements described with respect to one aspect or embodiment of this disclosure should be considered to be covered by other aspects or embodiments of this disclosure. Furthermore, the recitation of claim elements in conjunction with specific independent claims supports the recitation of those elements in conjunction with other independent claims. Throughout the disclosure and claims, when a composition or method is described as having, including, or comprising specific elements, compositions substantially consisting of, consisting of, or not containing the listed elements are also disclosed. All references cited herein are incorporated herein by reference.

Claims

1. A composition comprising a cyclic peptide selectively targeting adipose tissue, wherein the cyclic peptide comprises an amino acid sequence differing from a sequence selected from SEQ ID NO: 1-161 by no more than three amino acids, wherein each amino acid difference is independently selected from an amino acid insertion, deletion, or substitution.

2. The composition of claim 1, wherein the cyclic peptide comprises an amino acid sequence selected from SEQ ID NO: 1-161.

3. The composition of claim 1, wherein the cyclic peptide comprises an amino acid sequence that differs from the sequence selected from SEQ ID NO: 1-3 by no more than three amino acids.

4. The composition of claim 1, wherein the cyclic peptide comprises an amino acid sequence selected from SEQ ID NO: 1-3.

5. The composition of claim 1, wherein the cyclic peptide comprises an amino acid sequence that differs from SEQ ID NO: 1 by no more than three amino acids.

6. The composition of claim 1, wherein the cyclic peptide comprises the amino acid sequence according to SEQ ID NO:

1.

7. The composition of claim 1, wherein the cyclic peptide comprises an amino acid sequence that differs from SEQ ID NO: 2 by no more than three amino acids.

8. The composition of claim 1, wherein the cyclic peptide comprises the amino acid sequence according to SEQ ID NO:

2.

9. The composition of claim 1, wherein the cyclic peptide comprises an amino acid sequence that differs from SEQ ID NO: 3 by no more than three amino acids.

10. The composition of claim 1, wherein the cyclic peptide comprises the amino acid sequence according to SEQ ID NO:

3.

11. The composition of any one of claims 1 to 10, wherein the adipose tissue is white adipose tissue (WAT).

12. A peptide conjugate comprising a cyclic peptide and an agent associated with the cyclic peptide, the cyclic peptide comprising an amino acid sequence differing from a sequence selected from SEQ ID NO: 1-161 by no more than three amino acids, wherein each amino acid difference is independently selected from an amino acid insertion, deletion, or substitution.

13. The peptide conjugate of claim 12, wherein the cyclic peptide comprises an amino acid sequence selected from SEQ ID NO: 1-161.

14. The peptide conjugate of claim 12, wherein the cyclic peptide comprises an amino acid sequence that differs from the sequence selected from SEQ ID NO: 1-3 by no more than three amino acids.

15. The peptide conjugate of claim 12, wherein the cyclic peptide comprises an amino acid sequence selected from SEQ ID NO: 1-3.

16. The peptide conjugate of claim 12, wherein the cyclic peptide comprises an amino acid sequence that differs from SEQ ID NO: 1 by no more than three amino acids.

17. The peptide conjugate of claim 12, wherein the cyclic peptide comprises the amino acid sequence according to SEQ ID NO:

1.

18. The peptide conjugate of claim 12, wherein the cyclic peptide comprises an amino acid sequence that differs from SEQ ID NO: 2 by no more than three amino acids.

19. The peptide conjugate of claim 12, wherein the cyclic peptide comprises the amino acid sequence according to SEQ ID NO:

2.

20. The peptide conjugate of claim 12, wherein the cyclic peptide comprises an amino acid sequence that differs from SEQ ID NO: 3 by no more than three amino acids.

21. The peptide conjugate of claim 12, wherein the cyclic peptide comprises the amino acid sequence according to SEQ ID NO:

3.

22. The peptide conjugate of any one of claims 12 to 21, wherein the pharmaceutical agent is a diagnostic agent or a therapeutic agent, optionally wherein the therapeutic agent targets a target according to Table 2.

23. The peptide conjugate of any one of claims 12 to 22, wherein the agent is an inhibitory nucleic acid.

24. The peptide conjugate of any one of claims 12 to 22, wherein the agent is an antisense oligonucleotide.

25. The peptide conjugate of claim 23, wherein the agent is siRNA or miRNA.

26. The peptide conjugate of claim 24, wherein the antisense oligonucleotide is a phosphorodiamide morpholino oligonucleotide (PMO) or peptide nucleic acid (PNA).

27. The peptide conjugate of any one of claims 12 to 22, wherein the agent is adeno-associated virus (AAV).

28. The peptide conjugate of any one of claims 12 to 22, wherein the pharmaceutical agent is a lipid nanoparticle (LNP).

29. The peptide conjugate of any one of claims 12 to 28, wherein the pharmaceutical agent is non-covalently associated with the cyclic peptide.

30. The peptide conjugate of any one of claims 12 to 28, wherein the pharmaceutical agent is covalently associated with the cyclic peptide.

31. The peptide conjugate of claim 30, wherein the pharmaceutical agent is directly covalently associated with the cyclic peptide.

32. The peptide conjugate of claim 30, wherein the pharmaceutical agent is indirectly covalently associated with the cyclic peptide.

33. The peptide conjugate of claim 32, wherein the pharmaceutical agent is indirectly covalently associated with the cyclic peptide via a linker.

34. The peptide conjugate of claim 33, wherein the linker comprises a thioether bond, a disulfide bond, an oxime, a thiazoline, a hydrazone, an amide bond, an azide bond, or a maleimide bond.

35. The peptide conjugate of claim 33 or 34, wherein the linker comprises C1-C 30 Alkyl, C2-C 20 Alkyl, C3-C 12 Alkyl, C6-C 12 Alkyl or C6 alkyl group.

36. The peptide conjugate of any one of claims 33 to 34, wherein the linker comprises a C6 alkylamine.

37. The peptide conjugate of any one of claims 33 to 36, wherein the linker is a cuttable linker.

38. A pharmaceutical composition comprising a peptide conjugate as described in any one of claims 12 to 37 and a pharmaceutically acceptable carrier.

39. A method of delivering a pharmaceutical agent to the adipose tissue of a subject, the method comprising administering to the subject a peptide conjugate according to any one of claims 12 to 37 or a pharmaceutical composition according to claim 38.

40. A method for diagnosing, preventing, and / or treating adipose tissue-related diseases, the method comprising administering to the subject a peptide conjugate according to any one of claims 12 to 37 or a pharmaceutical composition according to claim 38, optionally wherein the disease is one of the diseases provided in Table 2.

41. The method of claim 40, wherein the disease is selected from obesity, cachexia, hyperglycemia, insulin resistance, type 2 diabetes, hypertension, cancer, heart disease, immune diseases, arthritis, diseases of the central nervous system, metabolic disorders, and age-related diseases.

Images