HK1168602A

HK1168602A - High penetration prodrug compositions of peptides and peptide-related compounds

Info

Publication number: HK1168602A
Application number: HK12106660.2A
Authority: HK
Inventors: 于崇曦; 徐丽娜; 陈玉华; 严彬冰; 涂仕前
Original assignee: 上海泰飞尔生化技术有限公司; 于崇曦
Priority date: 2009-05-08
Filing date: 2010-05-10
Publication date: 2013-01-04

Description

High penetration prodrug compositions of polypeptides and polypeptide-related compounds

Cross reference to related applications

The present invention is a continuation-in-part application of the united states patent application claiming priority of U.S. patent application No. 12/463,374 filed on 8.5.2009, which is incorporated by reference herein. And claim application priority of chinese patent application No. 200910135997.0 filed on 8/5/2009, which is incorporated by reference into this application.

Technical Field

The present invention relates to pharmaceutical compositions that rapidly penetrate one or more biological membrane barriers, and methods of using the compositions for preventing, diagnosing, and/or treating conditions or diseases in humans or animals treatable by polypeptides and related compounds. The invention also relates to a method for screening candidate new drugs and diagnosing biological individual symptoms by using the pharmaceutical composition. And a method for synthesizing the polypeptides HPPs/HPCs from N-terminal to C-terminal was developed.

Technical Field

Polypeptides are polymers composed of amino acids linked by amide bonds. Polypeptides play a variety of roles in biological systems. For example, polypeptide hormones are the largest group of hormones that regulate various biological processes in an organism. Injection of 1 ng of thyroid stimulating hormone releasing hormone into mice increases the relative thyroid action on blood iodideUptake (R.L.Kisliuk, Principles of Medicinal Chemistry, 4th Ed., W.O.Foye, et al Eds., Williams&Wilkins, 4th Ed.1995, p.606). Deglutition promoters (Tuftsin, threonine-lysine-proline-arginine) stimulate phagocytosis and promote antibody-dependent cytotoxicity (v.a. najjar, mol.cell.biochem.41, 1, 1981); methionine enkephalin (tyrosine-glycine-phenylalanine-methionine) was isolated from the brain and small intestine and functions the same as morphine in that it binds to the same receptor and has analgesic effects (j.r. jaffe and w.r. martin, in Pharmacological Basis of therapeutics, a.g. gilman, et al, eds., New York, Pergamon Press, 1990, p.481). Other polypeptide hormones include, but are not limited to, oxytocin (Pierce et al, j.199929, 1952), vasopressin (Kamm et al, j.am. chem. soc.50573, 1928), angiotensin (J.C. Garrison and M.J.Peach, in Pharmacological Basis of Therapeutics, A.G.Gilman, et al, eds., New York, Pergamon Press, 1990, p.749), gastrin (P.C. Emson and B.EB.Sandberg, Annu, Rep.Med.Chem.,1831, 1983), somatostatin (a.v. schally, et al, annu.rev.biochem.,4789, 1978), dynorphin (m.g. weisskopf, et al, Nature,362423, 1993), endothelin (a.m.doherty, j.med.chem.,351493, 1992), secretin (E.Jorper, Gastroenterology,55157, 1968), calcitonin (m.v.l.ray, et al, Biotechnology,1164, 1993), insulin (f.sanger, br.med.bull.,16183, 1960), and performance stimulating peptoids (CSP), and the like.

Another class of polypeptides is the antibacterial class of polypeptides, which are found to be involved in natural immunity in a variety of organisms (Reddy et al 2004). Because of the potential role of such polypeptides, as well as others, in the treatment of infectious diseases, much attention has been drawn, particularly because they are often effective against bacteria that have become resistant to conventional antibiotics. One well-known class of antimicrobial peptides is the tachypleins (tachplesins). Another class of antimicrobial peptides are the group peptides and their derivatives. Another class of antimicrobial peptides is hepcidin (hepcidin), also known as LEAP-1, which is an antimicrobial peptide used for liver expression.

Another class of polypeptides are calcium binding peptides that bind to calcified surfaces. An example of a calcium binding peptide comprises a three amino acid repeat (X-Y-Z)_nWherein X is aspartic acid, glutamic acid, asparaginic acid, alanine or glutamine, Y and Z are alanine, serine, threonine, phosphoserine or phosphothreonine, and n is any of numbers 1 to 40.

Unfortunately, polypeptides and polypeptide-related compounds are rapidly degraded by polypeptide proteases. When polypeptides and polypeptide-related compounds are administered orally, they are broken down within minutes. Other routes of administration of polypeptides and polypeptide-related compounds often result in pain and often require costly and frequent medical visits to treat chronic conditions.

Thus, there is a need for a novel composition that can be delivered to the site of action of a condition (e.g., a disease) quickly and efficiently to prevent, alleviate or treat the condition with minimal side effects.

Disclosure of Invention

In one aspect, the present invention provides a High Penetration pro-drug (HPP) or High Penetration Composition (HPC) comprising a functional unit covalently linked to a transport unit via a linker. The terms "HPP" and "HPC" may be used herein individually or collectively and are interchangeable unless otherwise indicated.

In certain embodiments, the functional unit of an HPP or HPC contains a portion (entity) of an agent (agent) for which rapid and efficient transport to a biological subject and/or transport across one or more biological barriers is desired

In certain embodiments, the functional units may be hydrophilic, lipophilic, or amphiphilic (i.e., both hydrophilic and lipophilic). For example, the lipophilicity of the functional unit may be intrinsic, or obtained by converting the hydrophilic portion of the functional unit to a lipophilic portion.

In certain instances, the functional unit of an HPP or HPC comprises a portion of a polypeptide or polypeptide-related compound. Polypeptide-related compound refers to a compound having a polypeptide structure, a peptide metabolite structure, or an agent that is metabolized into a polypeptide or polypeptide metabolite after HPP or HPC crosses one or more biological membrane barriers; polypeptide-related compounds may further refer to analogs or mimetics of a polypeptide or polypeptide metabolite, or agents that are metabolized into analogs or mimetics of a polypeptide or polypeptide metabolite after the HPP or HPC crosses one or more biological membrane barriers. Examples of polypeptides include, but are not limited to, polypeptide hormones (such as thyroid stimulating hormone releasing hormone, deglutition stimulating hormone (threonine-lysine-proline-arginine), methionine enkephalin (tyrosine-glycine-phenylalanine-methionine), oxytocin, angiotensin, gastrin, somatostatin, dynorphin, endothelin, secretin, calcitonin, insulin), enterostatin (such as valine-proline-aspartic acid-proline-arginine (VPDPR), valine-proline-glycine-proline-arginine (VPGPR), and alanine-proline-glycine-proline-Arginine (APGPR)), melanocortin II (loop (1, 6) -acetyl-norleucine-aspartic acid-histidine-phenylalanine-gpr) alanine-arginine-tryptophan-lysine-OH), opioid peptides (e.g., methionine enkephalin (H-tyrosine-glycine-phenylalanine-methionine-OH)), leucine-enkephalin (H-tyrosine-glycine-phenylalanine-leucine-OH), H-tyrosine-D-alanine-glycine-N-methyl-phenylalanine-methionine (O) -OL, and H-tyrosine-D-alanine-glycine-phenylalanine-leucine-OH), neuropeptides, alkaloids, anti-inflammatory polypeptides, antibacterial polypeptides (e.g., potency-stimulating peptides, naproxen, ghrelin, Group peptides and derivatives thereof), calcium binding-like polypeptides, regulatory-like polypeptides, peptide vaccines, and peptide mimetics (e.g., alpha-helix mimetics and beta-sheet mimetics).

In certain embodiments, the transport unit of the HPP or HPC contains a protonatable amino group, which may facilitate or enhance the transport of the HPP or HPC through or across one or more biological barriers. In certain embodiments, the protonatable amino group may be substantially protonated at the pH conditions of the biological barrier penetrated by the HPP or HPC. In certain embodiments, the amino group may be reversibly protonated or deprotonated.

In certain embodiments, a linker may covalently link the functional unit and the transport unit of an HPP or HPC, the linker comprising a bond that can be cleaved upon passage of the HPP or HPC through one or more biological barriers. The cleavable bond includes, for example, a covalent bond, an ether bond, a thioether bond, an amide bond, an ester bond, a thioester bond, a carbonate bond, a carbamate bond, a phosphate bond, or an oxime bond.

Another aspect of the invention relates to a pharmaceutical composition comprising an HPP or HPC of at least one polypeptide or polypeptide-related compound and a pharmaceutically acceptable carrier.

Another aspect of the invention relates to a method of crossing a biological barrier using an HPP or HPC of a polypeptide or polypeptide-related compound.

Another aspect of the invention relates to methods of diagnosing the onset, development, or slowing of a condition in a biological subject using HPPs or HPCs of a polypeptide or polypeptide-related compound. In some embodiments, the HPP (or HPC) or functional unit thereof may be tested. In certain embodiments, the HPP (or HPC) or functional unit thereof may itself be detectable, or labeled or conjugated with a detectable label.

Another aspect of the invention relates to methods of screening functional units, linkers, or transport units for a desired property.

Another aspect of the invention relates to a method of preventing, alleviating, or treating a condition in a biological subject by administering to the biological subject a composition of the invention. In certain embodiments, the methods relate to methods of treating a condition treatable by a polypeptide or polypeptide-related compound in a biological subject by administering to the subject a therapeutically effective dose of an HPP/HPC of the polypeptide or polypeptide-related compound, or a pharmaceutical composition thereof. In certain embodiments, conditions treatable by the method include, but are not limited to, pain, injury, inflammation-related conditions, microorganism-related conditions, neuropeptide-related conditions, hormone-related conditions, tumors, blood pressure abnormalities, obesity, brain damage, allergies, male and female sexual dysfunction, metastasis, and other conditions associated with deglutition, prenatal, postpartum, anti-AD activity, anti-diuretic activity, calcium balance, melanocytes, hormone release, platelet aggregation, central nervous system activity, and phagocytosis.

In certain embodiments, the pharmaceutical composition of HPP or HPC may be administered to a biological subject by a variety of routes, including, but not limited to, oral, intestinal, oral, nasal, topical, rectal, vaginal, aerosol, transmucosal, epithelial, transdermal, ocular, pulmonary, subcutaneous, and/or injectable routes of administration. In certain preferred embodiments, the pharmaceutical composition of HPP may be administered by oral, transdermal, topical, subcutaneous and/or injectable routes.

Consistent with the advantages of the present invention, and not intended to be limited by any particular mechanism, a therapeutically effective dose of HPP or HPC may be administered locally at the site of symptoms at a lower dose and to achieve a higher concentration. Advantages of the present invention also include, for example, avoidance of systemic administration, reduction of side effects (e.g., injection pain, gastrointestinal or renal reactions, or other side effects), and potential new therapeutic modalities due to high local concentrations of HPP, HPC, or active agents. Advantages of the present invention further include, for example, systemic administration of HPP or HPC to a biological subject can achieve bioavailability more quickly and efficiently, penetration of difficult to cross biological barriers (e.g., the blood-brain barrier), and new therapeutic approaches due to crossing biological barriers.

Description of the figures

FIG. 1: structural formula 2, structural formula 3, structural formula 4, structural formula 5, structural formula 6, structural formula 7, structural formula 8, structural formula 9, structural formula 10, structural formula 11, structural formula 12, structural formula 13, structural formula 14, structural formula 15, structural formula 16, structural formula 17, structural formula 18, structural formula 19, structural formula 20, structural formula 21, structural formula 22, structural formula 23, structural formula 24, structural formula 25, structural formula 26, structural formula 27, structural formula 28, structural formula 29, structural formula 30, structural formula 31, structural formula 32, structural formula 33, structural formula 34, structural formula 35, structural formula 36, structural formula 37, structural formula 38, structural formula 39, structural formula 40, structural formula 41, structural formula 42, structural formula 43, structural formula 45, structural formula 46, structural formula 47, structural formula 48, structural formula 49, structural formula 50, structural formula 51, structural formula 52, structural formula 53, structural formula 54, The structural formula 55, the structural formula 56, the structural formula 57, the structural formula 58, the structural formula 59, the structural formula 60, the structural formula 61, the structural formula 62, the structural formula 63, the structural formula 64, the structural formula 65, the structural formula 66, the structural formula 67, the structural formula 68, the structural formula 69, the structural formula 70, the structural formula 71, the structural formula 72, the structural formula 73, the structural formula 74, the structural formula 75, the structural formula 76, the structural formula 77, the structural formula 78, the structural formula 79, the structural formula 80, the structural formula 81, the structural formula 82, the structural formula 83, the structural formula 84, the structural formula 85, the structural formula 86, the structural formula 87, the structural formula 88, the structural formula 89, the structural formula 90, the structural formula 91, the structural formula 92, the structural formula 93, the structural formula 94, the structural formula 95, the structural, The structural formula 107, the structural formula 108, the structural formula 109, the structural formula 110, the structural formula 111, the structural formula 112, the structural formula 113, the structural formula 114, the structural formula 115, the structural formula 116, the structural formula 117, the structural formula 118, the structural formula 119, the structural formula 120, the structural formula 121, the structural formula 122, the structural formula 123, the structural formula 124, the structural formula 125, the structural formula 126, the structural formula 127, the structural formula 128, the structural formula 129, the structural formula 130, the structural formula 131, the structural formula 132, the structural formula 133, the structural formula 134, the structural formula 135, the structural formula 136, the structural formula 137, the structural formula 138, the structural formula 139, the structural formula 140, the structural formula 141, the structural formula 142, the structural formula 143, the structural formula 144, the structural formula 145, the structural formula 146, the structural formula 147, the structural, Formula 159, formula 160, formula 161, formula 162, formula 163, formula 164, formula 165, formula 166, formula 167, formula 168, formula 169, formula 170, formula 171, formula 172, formula 173, formula 174, formula 175, formula 176, formula 177, formula 178, formula 179, formula 180, formula 181, formula 182, formula 183, formula 184, formula 185, formula 186, formula 187, formula 188, formula 189, formula 190, formula 191, formula 192, formula 193, formula 194, formula 195, formula 196, formula 197, formula 198, formula 199, formula 200, formula 201, formula 202, formula 203, formula 204, formula 205, formula 206, formula 207, formula 208, formula 209, formula 210, formula 165, formula 166, formula, Structural formula 211, structural formula 212, structural formula 213, structural formula 214, structural formula 215, structural formula 216, structural formula 217, structural formula 218, structural formula 219, structural formula 220, structural formula 221, structural formula 222, structural formula 223, structural formula 224, structural formula 225, structural formula 226, structural formula 227, structural formula 228, structural formula 229, structural formula 230, structural formula 231, structural formula 232, structural formula 233, structural formula 234, structural formula 235, structural formula 236, structural formula 237, structural formula 238, structural formula 239, structural formula 240, structural formula 241, structural formula 242, structural formula 243, structural formula 244, structural formula 245, structural formula 246, structural formula 247, structural formula 248, structural formula 249, structural formula 250, structural formula 251, structural formula 252, structural formula 253, structural formula 254, structural formula 255, structural formula 256, structural formula 257, structural formula 258, structural formula 259, structural formula 260, structural formula 261, structural formula 262, Structural formula 263, structural formula 264, structural formula 265, structural formula 266, structural formula 267, structural formula 268, structural formula 269, structural formula 270, structural formula 271, structural formula 272, structural formula 273, structural formula 274, structural formula 275, structural formula 276, structural formula 277, structural formula 278, structural formula 279, structural formula 280, structural formula 281, structural formula 282, structural formula 283, structural formula 284, structural formula 285, structural formula 286, structural formula 287, structural formula 288, structural formula 289, structural formula 290, structural formula 291, structural formula 292, structural formula 293, structural formula 294, structural formula 295, structural formula 296, structural formula 297, structural formula 298, structural formula 299, structural formula 300, structural formula 301, structural formula 302, structural formula 303, structural formula 304, structural formula 305, structural formula 306, structural formula 307, structural formula 308, structural formula 309, structural formula 310, structural formula 311, structural formula 312, structural formula 313, structural formula 314, The structural formula 315, the structural formula 316, the structural formula 317, the structural formula 318, the structural formula 319, the structural formula 320, the structural formula 321, the structural formula 322, the structural formula 323, the structural formula 324, the structural formula 325, the structural formula 326, the structural formula 327, the structural formula 328, the structural formula 329, the structural formula 330, the structural formula 331, the structural formula 332, the structural formula 333, the structural formula 334 and the structural formula 335.

FIG. 2: acetyl-tyrosine (acetyl) -glycine-phenylalanine-methionine-diethylaminoethyl ester hydrochloride, N-dimethylaminobutylcarbonyl-tyrosine (acetyl) -glycine-phenylalanine-methionine-butyl ester hydrochloride, cyclo (1, 6) -acetyl-norleucine-aspartic acid-histidine-phenylalanine-arginine (diacetyl) -tryptophan-lysine-diethylaminoethyl ester hydrochloride, cyclo (1, 6) -norleucine-aspartic acid-histidine-D-phenylalanine (4-iodo) -arginine (acetyl) -tryptophan-lysinamide hydrochloride, arginine hydrochloride, glycine-phenylalanine-methionine-methyl ester hydrochloride, arginine hydrochloride, glycine-phenylalanine-methionine-arginine hydrochloride, and salts thereof, Cyclo (1, 6) -norleucine-aspartic acid-histidine-D-alanine (2-naphthyl) -arginine (nitro) -tryptophan-lysinamide hydrochloride, acetyl-valine-proline-glycine-proline-arginine (diacetyl) -diethylaminoethyl ester hydrochloride, acetyl-tyrosine-glycine-phenylalanine-methionine, cyclo (1, 6) -acetyl-norleucine-aspartic acid-histidine-phenylalanine-arginine-tryptophan-lysine, and cyclo (1, 6) -acetyl-norleucine-aspartic acid-histidine-D-phenylalanine (4-iodo) -arginine-tryptophan-lysine And the cumulative amount of valine-proline-glycine-proline-arginine that passes through human skin tissue in Franz cells (n ═ 5). Wherein each group of media is phosphate buffer (0.2M) with pH 7.4.

Disclosure of Invention

I. Of High Penetration Prodrugs (HPPs) or High Penetration Compositions (HPCs) of polypeptides or polypeptide-related compounds And (5) structure.

One aspect of the present invention relates to a High Penetration Prodrug (HPP) or a High Penetration Composition (HPC). "high penetrating prodrug" or "HPP" or "high penetrating composition" or "HPC" refers herein to a composition that contains a functional unit and a transport unit covalently linked by a linker.

The functional unit of an HPP or HPC contains a portion of the parent drug (motif), which has the following properties: 1) it is desirable to deliver the parent drug or HPP/HPC into a biological subject and/or to transport the parent drug across a biological barrier; 2) HPP/HPC can penetrate or cross biological barriers; and 3) HPP/HPC may be cleaved such that a portion of the parent drug is converted to the parent drug or a metabolite of the parent drug.

In certain embodiments, the functional units may be hydrophilic, lipophilic, or amphiphilic (i.e., both hydrophilic and lipophilic). The lipophilic part of the functional unit may be intrinsic or obtained by converting one or more hydrophilic parts of the functional unit into a lipophilic part. For example, the lipophilic portion of the functional unit can be converted to a lipophilic group by organic synthesis by converting one or more hydrophilic groups of the functional unit. Examples of hydrophilic groups include, but are not limited to, carboxyl, hydroxyl, thiol, amino, phosphate/phosphonate, and carbonyl. Lipophilic moieties generated by modification of these hydrophilic groups include, but are not limited to, ethers, thioethers, esters, thioesters, carbonates, carbamates, amides, phosphates, and oximes. In certain embodiments, the functional unit may be lipophilic by acetylation. In certain embodiments, the functional units may be lipophilic by esterification.

In certain embodiments, the parent drug for HPP or HPC is selected from the group consisting of polypeptides and polypeptide-related compounds. A portion of the polypeptide and related compounds may be further converted to lipophilic as previously described. In the present invention, the term "polypeptide HPP/HPC" refers to a High Penetration Prodrug (HPP) or High Penetration Composition (HPC) of a polypeptide or polypeptide-related compound.

Polypeptides are well known and used for a variety of conditions. In the present invention, a polypeptide refers to an amino acid sequence having a length of about 2 to 50 amino acids. For example, a polypeptide may consist of 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 or more amino acids. The polypeptide may also comprise D-amino acids and/or L-amino acids.

Amino acids are a class of compounds containing both amino and carboxyl functional groups. The carbon atom immediately adjacent to the carbonyl group of the carboxyl function is referred to as the alpha carbon. Amino acids with a branch linked to an alpha carbon are referred to as alpha amino acids. Where the amino acid has an amino group and a carbon chain attached to the alpha carbon, the carbon atoms may be designated as alpha, beta, and gamma carbons in that order from the carbonyl carbon. Amino acids in which the amino group is linked to the beta or gamma carbon are referred to as beta or gamma amino acids, respectively, and so on.

Alpha amino acids are a class of amino acids in which the amino group and the carboxylic acid group are attached to the same carbon atom (alpha carbon). The alpha carbon is the first carbon atom from the carboxylic acid group. The alpha amino acid has the structure of formula 1:

H₂NCHR′COOH

structural formula 1

Including stereoisomers and pharmaceutically acceptable salts thereof, wherein R is selected from the group consisting of substituted and unsubstituted imidazolyl, substituted and unsubstituted guanidino, substituted and unsubstituted carboxy, substituted and unsubstituted amide, substituted and unsubstituted alkyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocycloalkyl, substituted and unsubstituted alkoxy, substituted and unsubstituted alkylthio, substituted and unsubstituted alkylamino, substituted and unsubstituted alkylcarbonyl, substituted and unsubstituted perfluoroalkyl, substituted and unsubstituted halocarbon, substituted and unsubstituted aryl, and substituted and unsubstituted heteroaryl.

In certain embodiments, the amino acid comprises a structure represented by structural formula 1, including stereoisomers and pharmaceutically acceptable salts thereof, wherein R' is selected from the group consisting of: H. CH (CH)₃-、HN＝C(NH₂)-NH-(CH₂)₃-、H₂N-CO-CH₂-、HOOC-CH₂-、HS-CH₂-、H₂N-CO-(CH₂)₂-、HS-(CH₂)₂-、HOOC-CH₂-、CH₃-CH₂-CH(CH₃)-、(CH₃)₂-CH-CH₂-、H₂N-(CH₂)₄-、CH₃-S-(CH₂)₂-、Phenyl-CH₂-、HO-CH₂-、CH₃-CH(OH)-、4-OH-Phenyl-CH₂-、CH₃-CH(CH₃)-、And derivatives thereof.

Examples of alpha amino acids include, but are not limited to, alanine (Ala), arginine (Arg), asparagine (Asn), aspartic acid (Asp), cysteine (Cys), glutamic acid (Glu), glutamine (Gln), glycine (Gly), histidine (His), homocysteine (Hcy), homoserine (Hse), isoleucine (Ile), leucine (Leu), lysine (Lys), methionine (Met), norleucine (Nle), norvaline (Nva), ornithine (Orn), penicillamine (Pen), phenylalanine (Phe), proline (Pro), serine (Ser), tyrosine (Thr), threonine (Trp), tryptophan (Tyr), valine (Val), pyroglutamic acid (pGLU), dinitrobenzyl lysine (dnp-LYS }, threonine phosphate (pTHR }, serine phosphate (pSER), tyrosine phosphate (pTYR) Citrulline (CIT), N-methylalanine (nme-ALA), N-methylisoleucine (nme-ILE), N-methylleucine (nme-LEU), N-methylphenylalanine (nme-PHE), N-methylvaline (nme-VAL), N-methylserine (nme-SER), N-methylthreonine (nme-THR), N-methyltyrosine (nme-TYR), alpha-aminobutyric acid (alpha-ABA), iso-aspartic acid (iso-ASP), acetyl lysine (Ac-LYS), 2-methylalanine (2-Me-ALA), and oxamic acid (OXA).

Beta amino acids are a class of amino acids in which the amino group is attached to the beta carbon (the second carbon atom from the carboxylic acid group), and include, but are not limited to, beta-alanine (beta-Ala), beta-arginine (beta-Arg), beta-asparagine (beta-Asn), beta-aspartic acid (beta-Asp), beta-cysteine (beta-Cys), beta-

Gamma amino acids are a class of amino acids in which the amino group is attached to the gamma carbon (the third carbon atom from the carboxylic acid group). Examples of gamma amino acids include, but are not limited to, gamma glutamic acid (gamma-GLU).

Polypeptide-related compounds are compounds that contain a polypeptide structure, a polypeptide metabolite structure, or are agents that are metabolized into a polypeptide or polypeptide metabolite when the polypeptide HPP or HPC crosses one or more biological barriers. Polypeptide-related compounds further include analogs or mimetics of the polypeptide or polypeptide metabolite, or agents that are metabolized to an analog or mimetic of the polypeptide or polypeptide metabolite upon crossing one or more biological barriers by the polypeptide HPP or HPC.

Examples of polypeptides and polypeptide-related compounds include, but are not limited to, polypeptide hormones, neuropeptides, alkaloids, antibacterial peptides, anti-inflammatory peptides, polypeptide toxins, regulatory peptides, calcium binding peptides, polypeptide vaccines and polypeptide mimetics.

Polypeptide hormones are a class of polypeptides that have endocrine function in living animals. Polypeptide hormones also play an important role in plant cell-to-cell communication and plant defense. Polypeptide hormones are produced by a variety of organs and tissues, such as the heart (atrial natriuretic peptide (ANP), atrial natriuretic peptide (ANF)), the pancreas (e.g., insulin, enterostatin, somatostatin), the gastrointestinal tract (cholecystokinin, gastrins (e.g., gastrin-34, gastrin-17, and gastrin-14), opioids (e.g., methionine-enkephalin, leucine-enkephalin, tyrosine-D-alanine-glycine-N-methyl-phenylalanine (oxy) -OL, and tyrosine-D-alanine-glycine-phenylalanine-leucine), cholecystokinin, secreted hormones, involuntains, vasoactive intestinal peptides, and glucagon), adipose tissue depots (e.g., leptin), luteinizing hormones, follitropin, prolactin, adrenocorticotropic hormone (ACTH), growth hormone, antidiuretic hormone, oxytocin, melanocortins (e.g., melanocortin II)), thyroid (e.g., calcitonin), spleen (deglutition promoter), brain (e.g., oxytocin, dynorphin), liver (e.g., angiotensin I and angiotensin II), and endothelial tissue (e.g., endothelin). Other examples of polypeptide hormones include, but are not limited to, thyroid-releasing hormone (TRH) and bradykinin.

Neuropeptides refer to polypeptides found in neural tissue that are involved in the control and signaling processes. Examples of neuropeptides include, but are not limited to, neurotransmitters (e.g. N-acetyl glutamate, gastrins, cholecystokinin, neuropeptide Y, posterior pituitary hormones, oxytocin, secretagogues, substance P, somatostatin, Vasoactive Intestinal Peptide (VIP), opioid peptides (e.g. enkephalin, dynorphin, endorphin), somatotropin, neurotensin, TRH, atrial peptides.

Alkaloids refer to polypeptides that are typically isolated from plants, fungi, and some animals such as crustaceans. Alkaloids participate in the defense of an organism against destruction by another organism. Examples of alkaloids include, but are not limited to, ergotamine, climax, dynorphin A- (1-8) -octapeptide, N beta- (D-leucine-D-arginine-D-leucine-D-phenylalanine) -cyprohexamine (naltrexamine).

Antimicrobial peptides refer to polypeptides that inhibit the growth of microorganisms, such as bacterial cells and topical fungi, as well as protozoa. Examples of antimicrobial peptides include, but are not limited to bacitracin, gramicidin, valinomycin, proficient peptides, tachypleins, histones and derivatives thereof.

Examples of anti-inflammatory peptides are the sequence Seq ID: 48, Seq ID: 49, and Seq ID: (Table A)

Polypeptide toxins refer to toxic polypeptides. Examples of polypeptide toxins are palutoxins, american spider toxins, and curratoxins.

Regulatory peptides refer to polypeptides that regulate one or more biological processes. Examples of regulatory polypeptides include, but are not limited to anserine (anserine) and carnosine.

Other examples of polypeptides and polypeptide-related compounds include calcium-binding peptides, polypeptide vaccines (e.g., p45(IEIGLEGKGFEPTLE ALFGK) and p210(KTTKQSFDLS VKAQY KKNKH)), and polypeptide mimetics (e.g., alpha-helix mimetics and beta-sheet mimetics).

In certain embodiments, the functional units of HPP/HPC of polypeptides and related compounds comprise the structure of structural formula F-1:

structural formula F-1

Including stereoisomers and pharmaceutically acceptable salts thereof, wherein:

each A₁-A_mEach independently selected from the group consisting of: 2-naphthylalanine, substituted and unsubstituted alkyl groups, substituted and unsubstituted cycloalkyl groups, substituted and unsubstituted heterocycloalkyl groups, substituted and unsubstituted alkoxy groups, substituted and unsubstituted alkenyl groups, substituted and unsubstituted alkynyl groups, substituted and unsubstituted aryl groups, substituted and unsubstituted heteroaryl groups, and structural formula a;

structural formula A

Each A₁-A_mP in (A) is independently selected from integers;

each A₁-A_mZ on each carbon atom of_A-1Each A₁-A_mZ on each carbon atom of_A-2，Z_NT，Z_CT-1And Z_CT-2Each independently selected from the group consisting of: h, CH₃，C₂H₅，C₃H₇，CF₃，C₂F₅，C₃F₇Substituted and unsubstituted hydrocarbon groups, substituted and unsubstituted polyfluoroalkyl groups, and substituted and unsubstituted halogenated hydrocarbon groups;

each A₁-A_mR on each carbon atom of_A，R_NTAnd R_CTEach independently selected from the group consisting of: h, substituted and unsubstituted imidazolyl, substituted and unsubstituted quaternidino, substituted and unsubstituted carboxy, substituted and unsubstituted carboxamide, substituted and unsubstituted alkyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocycloalkyl, substituted and unsubstituted alkoxy, substituted and unsubstituted alkylthio, substituted and unsubstituted alkylamino, substituted and unsubstituted hydrocarbonyl, substituted and unsubstituted polyfluoroalkyl, substituted and unsubstituted haloalkyl, substituted and unsubstituted aryl, and substituted and unsubstituted heteroaryl;

when a is one₁-A_mR on each carbon atom when p is an integer of not less than 2_AZ, which may be identical or different, on each carbon atom_A-1May be the same or different;

the amino and carboxyl groups on the polypeptide chain may further form lactam bridges; and

thiols can further form disulfide bridges.

In certain embodiments, the functional units of the polypeptide HPP/HPC comprise a structure represented by structural formula F-1 as previously defined, including stereoisomers and pharmaceutically acceptable salts thereof, wherein A₁-A_mR of (A) to (B)_AMay be further lipophillized by acetylation or esterification.

In certain embodiments, the functional unit of the polypeptide HPP/HPC comprises a structure represented by structural formula F-1 as previously defined, including stereoisomers and pharmaceutically acceptable salts thereof, wherein m is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12.

In certain embodiments, the functional units of HPPs of the polypeptides and polypeptide-related compounds comprise a structure represented by structural formula F-1 as previously defined, including stereoisomers and pharmaceutically acceptable salts thereof, wherein p is 1, 2 or 3.

In certain embodiments, the functional units of HPPs of the polypeptides and polypeptide-related compounds comprise a structure represented by structural formula F-1 as previously defined, including stereoisomers and pharmaceutically acceptable salts thereof,

wherein:

p is 1, 2 or 3;

each A₁-A_mZ on each carbon atom of_A-1Each A₁-A_mZ on each carbon atom of_A-2，Z_NT，Z_CT-1And Z_CT-2Each independently selected from the group consisting of: h, CH₃，C₂H₅，C₃H₇，CF₃，C₂F₅，C₃F₇Substituted and unsubstituted hydrocarbon groups, substituted and unsubstituted cycloalkyl groups, substituted and unsubstituted heterocycloalkyl groups, substituted and unsubstituted polyfluoroalkyl groups, and substituted and unsubstituted halogenated hydrocarbon groups;

each A₁-A_mR on each carbon atom of_A，R_NTAnd R_CTEach independently selected from the group consisting of: h, substituted and unsubstituted imidazolyl, substituted and unsubstituted quinodino, substituted and unsubstituted carboxyl, substituted and unsubstituted carboxamide, substituted and unsubstituted alkyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocycloalkyl, substituted and unsubstituted alkoxy, and substituted and unsubstituted alkoxySubstituted and unsubstituted hydrocarbylthio, substituted and unsubstituted hydrocarbylamino, substituted and unsubstituted hydrocarbylcarbonyl, substituted and unsubstituted polyfluoroalkyl, substituted and unsubstituted haloalkyl, substituted and unsubstituted aryl, and substituted and unsubstituted heteroaryl;

thiols can further form disulfide bridges.

In the present invention, "pharmaceutically acceptable salt" refers to a salt of the compound of the present invention which can be safely used in a subject. Pharmaceutically acceptable salts include salts of acidic or basic groups present in the compounds of the invention. Pharmaceutically acceptable acid addition salts include, but are not limited to, hydrochloride, hydrobromide, hydrogen iodide, nitrate, sulfate, hydrogen sulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, citrate, tartrate, pantothenate, acid tartrate, ascorbate, succinate, maleate, gentiopiconate, fumarate, gluconate, glucuronate, gluconate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and pamoate (e.g., 1, 11-methylene-bis- (2-hydroxy-3-naphthoate)). Certain compounds of the invention may form pharmaceutically acceptable salts with a variety of amino acids. Suitable basic salts include, but are not limited to, aluminum, calcium, lithium, magnesium, potassium, sodium, zinc, and diethanolamine salts. For an overview of pharmaceutically acceptable salts, see BERGEET AL., 66J. PHARM. SCI.1-19(1977), incorporated herein by reference.

In the present invention, unless otherwise specified, the term "hydrocarbon group" means a branched or unbranched, saturated or unsaturated, monovalent or polyvalent hydrocarbon group, and includes saturated hydrocarbon groups, alkenyl groups, and alkynyl groups. Examples of hydrocarbyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, vinyl, propenyl, butenyl, isobutenyl, pentenyl, hexenyl, heptenyl, octenyl, nonenyl, decenyl, undecenyl, dodecenyl, ethynyl, propynyl, butynyl, isobutenyl, pentynyl, hexynyl, heptynyl, octynyl, nonynyl, decynyl, undecenyl, dodecenyl, methylene, ethylene (ethylene), propylene (propylene), isopropylene (isopropylene), butylene (butylene), isobutylene (isobutylene), t-butylene (t-butylene), pentylene (pentylene), hexylene (heptylene), heptylene (heptylene), octylene (octylene), nonylene (nonylene), Decylene (decylene), undecylene (undecylene), and dodecylene (dodecylene). In certain embodiments, the hydrocarbon group contains from 1 to 30 carbon atoms. In certain embodiments, the hydrocarbon group contains 1 to 20 carbon atoms. In certain embodiments, the hydrocarbon group contains 1 to 12 carbon atoms.

In the present invention, unless otherwise specified, the term "cyclic hydrocarbon group" means a hydrocarbon group containing at least one ring and not containing an aromatic ring. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl, and cyclododecyl. In certain embodiments, the hydrocarbon group contains from 1 to 30 carbon atoms. In certain embodiments, the hydrocarbon group contains 1 to 20 carbon atoms. In certain embodiments, the hydrocarbon group contains 1 to 12 carbon atoms.

In the present invention, unless otherwise specified, the term "heterocyclic hydrocarbon group" means a cyclic hydrocarbon group in which at least one of the ring atoms is not a carbon atom. Non-carbon ring atoms include, but are not limited to, sulfur, oxygen, and nitrogen.

In the present invention, unless otherwise specified, the term "hydrocarbyloxy" refers to a hydrocarbyl, cycloalkyl or heterocycloalkyi group containing one or more oxygen atoms. Examples of hydrocarbyloxy groups include, but are not limited to, -CH₂-OH、-OCH₃-O-hydrocarbyl, -hydrocarbyl-OH, -hydrocarbyl-O-hydrocarbyl-, wherein the two hydrocarbyl groups may be the same or different.

In the present invention, unless otherwise specified, the term "halogenated hydrocarbon group" means a hydrocarbon group, a cycloalkyl group or a heterocyclic hydrocarbon group containing one or more halogen atoms which may be the same or different. The term "halogen" refers to fluorine, chlorine, bromine or iodine. Examples of halogenated hydrocarbyl groups include, but are not limited to, -hydrocarbyl-F, -hydrocarbyl-Cl, -hydrocarbyl-Br, -hydrocarbyl-I, -hydrocarbyl (F) -, -hydrocarbyl (Cl) -, -hydrocarbyl (Br) -, and-hydrocarbyl (I) -.

In the present invention, unless otherwise specified, the term "alkylthio" refers to a hydrocarbon, cycloalkyl or heterocyclic hydrocarbon group containing one or more sulfur atoms. Examples of hydrocarbylthio groups include, but are not limited to, -CH₂-SH、-SCH₃-S-hydrocarbyl, -hydrocarbyl-SH, -hydrocarbyl-S-hydrocarbyl-, wherein the two hydrocarbyl groups may be the same or different.

In the present invention, unless otherwise specified, the term "hydrocarbylamino" refers to a hydrocarbyl, cycloalkyl or heterocycloalkyl group containing one or more nitrogen atoms. Examples of hydrocarbylamino groups include, but are not limited to, -CH₂-NH，-NCH₃-N (hydrocarbyl) -hydrocarbyl, -N-hydrocarbyl, -hydrocarbyl-NH₂-hydrocarbyl-N-hydrocarbyl and-hydrocarbyl-N (hydrocarbyl) -hydrocarbyl, wherein the hydrocarbyl groups may be the same or different.

In the present invention, unless otherwise specified, the term "alkyl (alkyl)" refers to a hydrocarbon, cycloalkyl or heterocyclic hydrocarbon group containing one or more carbonyl groups. Examples of hydrocarbon carbonyl groups include, but are not limited to, aldehyde groups (R-C (O) -H), ketone groups (-R-C (O) -R '), carboxylic acid groups (R-COOH), ester groups (-R-COO-R'), carboxamide groups, (-R-COO-N (R ') R'), enol groups (-R-C (O) -C (R ') - [ C (R') R '), acyl halide groups (-R-C (O) -X), and anhydride groups (-R-C (O) - [ O ] (-C (O)) -R'), wherein R, R 'and R' may be the same or different hydrocarbon, cyclic or heterocyclic hydrocarbon groups.

In the present invention, unless otherwise specified, the term "polyfluoroalkyl" refers to a hydrocarbon, cycloalkyl or heterocyclic hydrocarbon group containing one or more fluoro groups, including, but not limited to, polyfluoromethyl, polyfluoroethyl, polyfluoropropyl.

In the present invention, the term "aryl" refers to a chemical structure containing one or more aromatic rings, unless otherwise specified. In certain embodiments, the ring atoms are all carbon atoms. In certain embodiments, one or more ring atoms are non-carbon atoms, such as oxygen atoms, nitrogen atoms, or sulfur atoms ("heteroaryl"). Examples of aryl groups include, but are not limited to, phenyl, benzyl, naphthyl, anthracenyl, pyridinyl, quinolinyl, isoquinolinyl, pyrazinyl, quinoxalinyl, acridinyl, pyrimidinyl, quinazolinyl, pyridazinyl, pyrimidinyl, pyridazinyl, cinnamyl, cinn,Cinnoline base(cinnolinyl), imidazolyl, benzimidazolyl, purinyl, indolyl, furyl, benzofuryl, isobenzofuryl, pyrrolyl, indolyl, isoindolyl, thienyl, benzothienyl, pyrazolyl, indazolyl, oxazolyl, benzoxazolyl, isoxazolyl, benzisoxazolyl, thiazolyl (thiaxolyl), guanidino (quaternino), and benzothiazolyl.

In certain embodiments, the transport unit of the HPP/HPC contains a protonatable amino group that facilitates the transport or crossing of the HPP/HPC across one or more biological barriers (e.g., at least about 10-fold, about 50-fold, about 100-fold, about 300-fold, about 500-fold, about 1000-fold higher than the parent drug). In certain embodiments, the protonatable amino group is largely protonated at physiological pH. In certain embodiments, the amino group can be reversibly protonated. In certain embodiments, the transport unit may or may not be cleaved from the functional unit after the HPP/HPC has crossed one or more biological barriers. In certain embodiments, the transport unit may be obtained from a functional unit, in particular a polypeptide or polypeptide-related compound containing at least one free amino group. In certain embodiments, when a polypeptide or polypeptide-related compound contains more than one protonatable group, the polypeptide or polypeptide-related compound is modified to leave only one or two protonatable amino groups, while all other protonatable groups are protected.

In certain embodiments, the protonatable amino group is selected from the group consisting of pharmaceutically acceptable substituted and unsubstituted primary amino groups, pharmaceutically acceptable substituted and unsubstituted secondary amino groups, and pharmaceutically acceptable substituted and unsubstituted tertiary amino groups.

In certain embodiments, the protonatable amino group is selected from the group consisting of formula Na, formula Nb, formula Nc, formula Nd, formula Ne, formula Nf, formula Ng, formula Nh, formula Ni, formula Nj, formula Nk, formula Nl, formula Nm, formula Nn, formula No, formula Np, formula Nq, and formula Nr:

structural formula Na structural formula Nb

Formula Nc formula Nd

Structural formula Ne structural formula Nf

Structural formula Ng structural formula Nh

Structural formula Ni structural formula Nj

Structural formula Nk structural formula Nl

Formula Nm formula Nn

Structural formula No structural formula Np

Structural formula Nq structural formula Nr

Including stereoisomers and pharmaceutically acceptable salts thereof

In the present invention, unless otherwise specified, each R is₁₁-R₁₆Each independently selected from the group consisting of: none, H, CH₂COOR₁₁Substituted and unsubstituted hydrocarbyl, substituted andunsubstituted cycloalkyl, substituted and unsubstituted heterocycloalkyl, substituted and unsubstituted aryl, substituted and unsubstituted heteroaryl, substituted and unsubstituted alkoxy, substituted and unsubstituted alkylthio, substituted and unsubstituted alkylamino, substituted and unsubstituted perfluoroalkyl, substituted and unsubstituted haloalkyl, wherein any carbon or hydrogen atom may be further replaced by O, S, P, NR₁₁Or other pharmaceutically acceptable groups;

in certain embodiments, the linking moiety covalently linking the functional unit and the transport unit of the HPP/HPC comprises a bond that can be cleaved after the HPP/HPC crosses one or more biological barriers. The linkage that can be cleaved includes, for example, a covalent bond, an ether linkage, a thioether linkage, an amide linkage, an ester linkage, a thioester linkage, a carbonate linkage, a carbamate linkage, a phosphate linkage or an oxime linkage.

In certain embodiments, the HPP/HPC of the polypeptide or polypeptide-related compound has the structure of structural formula L-1:

structural formula L-1

Including stereoisomers and pharmaceutically acceptable salts thereof, wherein:

f is a functional unit of HPP/HPC, a polypeptide and polypeptide-related compound. Examples of F include structural formula F-1 as previously defined;

T_cand T_NIndependently selected from the group consisting of none, H, substituted and unsubstituted hydrocarbyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocycloalkyl, substituted and unsubstituted hydrocarbyloxy, substituted and unsubstituted alkenyl, substituted and unsubstituted alkynyl, substituted and unsubstituted aryl, substituted and unsubstituted heteroaryl, as defined previously, formula Na, formula Nb, formula Nc, formula Nd, formula Ne, formula Nf, formula Ng, formula Nh, formula Ni, formula Nj, formula Nk, formula Nl, formula Nm, formula Nn, formula No, formula Np, formula Nq, and formula Nr;

L_1Cand L_1NAre independently selected from O, S, -N (L) and₃)-，-N(L₃)-CH₂-O，-N(L₃)-CH₂-N(L₅)-，-O-CH₂-O-，-O-CH(L₃) -O and-S-CH (L)₃) -O-composition set;

L_2Cand L_2NAre independently selected from O, S, -N (L) and₃)-，-N(L₃)-CH₂-O，-N(L₃)-CH₂-N(L₅)-，-O-CH₂-O-，-O-CH(L₃)-O，-S-CH(L₃)-O-，-O-L₃-，-N-L₃-，-S-L₃-，-N(L₃)-L₅-and L₃A set of compositions;

L_4Cand L_4NAre each independently selected from the following groups C ═ O, C ═ S,a set of compositions;

each L_1C，L_1N，L_2C，L_2N，L_4CAnd L_4N，L₃And L₅Each independently selected from the group consisting of: zero (no atom), H, CH₂COOL₆Substituted and unsubstituted alkyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocycloalkyl, substituted and unsubstituted aryl, substituted and unsubstituted heteroaryl, substituted and unsubstituted alkoxy, substituted and unsubstituted alkylthio, substituted and unsubstituted alkylamino, substituted and unsubstituted perfluoroalkyl, substituted and unsubstituted haloalkyl, wherein any carbon or hydrogen atom may be further replaced by O, S, P, NL₃Or other pharmaceutically acceptable groups;

L₆is independently selected from the group consisting of: h, OH, Cl, F, Br, I, substituted and unsubstitutedSubstituted hydrocarbyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocycloalkyl, substituted and unsubstituted aryl, substituted and unsubstituted heteroaryl, substituted and unsubstituted alkoxy, substituted and unsubstituted alkylthio, substituted and unsubstituted alkylamino, substituted and unsubstituted perfluoroalkyl, substituted and unsubstituted haloalkyl, wherein any carbon or hydrogen atom may be further replaced by O, S, N, P (O) OL₆，CH＝CH，C≡C，CHL₆，CL₆L₇Aryl, heteroaryl or cyclyl substituted; and

L₇is selected from the group consisting of: h, OH, Cl, F, Br, I, substituted and unsubstituted hydrocarbyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocycloalkyl, substituted and unsubstituted aryl, substituted and unsubstituted heteroaryl, substituted and unsubstituted hydrocarbyloxy, substituted and unsubstituted hydrocarbylthio, substituted and unsubstituted hydrocarbylamino, substituted and unsubstituted perfluoroalkyl, substituted and unsubstituted halohydrocarbyl, wherein any carbon or hydrogen atom may be further replaced by O, S, N, P (O) OL₆，CH＝CH，C≡C，CHL₆，CL₆L₇Aryl, heteroaryl or cyclyl substituted.

In certain embodiments, the HPP or HPC of a polypeptide or polypeptide-related compound comprises a structure represented by structural formula L-1, including stereoisomers and pharmaceutically acceptable salts thereof, wherein:

F，L_1C，L_1N，L_2C，L_2N，T_Cand T_NAs previously defined; and

L_4Cand/or L_4NIs C ═ O.

F，L_1C，L_1N，L_2C，L_2N，L_4Cand L_4NAs previously defined;

T_Cis the transport unit for the polypeptide HPP/HPC. E.g. T_CIs selected from the group consisting of: structural formula Na, structural formula Nb, structural formula Nc, structural formula Nd, structural formula Ne, structural formula Nf, structural formula Ng, structural formula Nh, structural formula Ni, structural formula Nj, structural formula Nk, structural formula Nl, structural formula Nm, structural formula Nn, structural formula No, structural formula Np, structural formula Nq and structural formula Nr;

T_Nis selected from the group consisting of absent, H, substituted and unsubstituted alkyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocycloalkyl, substituted and unsubstituted alkoxy, substituted and unsubstituted alkenyl, substituted and unsubstituted alkynyl, substituted and unsubstituted aryl, and substituted and unsubstituted heteroaryl.

In certain embodiments, the HPP or HPC of a polypeptide or related compound comprises a structure represented by structural formula L-1, including stereoisomers and pharmaceutically acceptable salts thereof, wherein:

F，L_1C，L_1N，L_2C，L_2N，L_4Cand L_4NAs previously defined;

T_Nis the transport unit for the polypeptide HPP/HPC. For example, TN is selected from the formula Na, formula Nb, formula Nc, formula Nd, formula Ne, formula Nf, formula Ng, formula Nh, formula Ni, formula Nj, formula Nk, formula Nl, formula Nm, formula Nn, formula No, formula Np, formula Nq and formula Nr as defined above; and

T_Cselected from the group consisting of none, H, substituted and unsubstituted alkyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocycloalkyl, substituted and unsubstituted alkoxy, substituted and unsubstituted alkenyl, substituted and unsubstituted alkynyl, substituted and unsubstituted aryl, and substituted and unsubstituted heteroaryl.

In certain embodiments, the polypeptide HPP/HPC has the following structural formula L-2:

structural formula L-2

Including stereoisomers and pharmaceutically acceptable salts thereof, wherein:

each A₁-A_mEach independently selected from the group consisting of: 2-naphthylalanine, substituted and unsubstituted alkyl groups, substituted and unsubstituted cycloalkyl groups, substituted and unsubstituted heterocycloalkyl groups, substituted and unsubstituted alkoxy groups, substituted and unsubstituted alkenyl groups, substituted and unsubstituted alkynyl groups, substituted and unsubstituted aryl groups, substituted and unsubstituted heteroaryl groups, structural formula a and structural formula B;

structural formula A structural formula B

Each A₁-A_mEach p of (a) is independently selected from integers;

each A₁-A_mT of_B，T_CAnd T_NIndependently selected from the group consisting of n, H, substituted and unsubstituted hydrocarbyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocycloalkyl, substituted and unsubstituted hydrocarbyloxy, substituted and unsubstituted alkenyl, substituted and unsubstituted alkynyl, substituted and unsubstituted aryl, substituted and unsubstituted heteroaryl, formula Na, formula Nb, formula Nc, formula Nd, formula Ne, formula Nf, formula Ng, formula Nh, formula Ni, formula Nj, formula Nk, formula Nl, formula Nm, formula Nn, formula No, formula Np, formula Nq, and formula Nr;

each A₁-A_mL of_1B，L_1CAnd L_1NEach independently selected from the group consisting ofAnd (3) gathering: none, O, S, -N (L)₃)-，-N(L₃)-CH₂-O，-N(L₃)-CH₂-N(L₅)-，-O-CH₂-O-，-O-CH(L₃) -O, and-S-CH (L)₃)-O；

Each A₁-A_mL of_2B，L_2CAnd L_2NEach independently selected from the group consisting of: none, O, S, -N (L)₃)-，-N(L₃)-CH₂-O，-N(L₃)-CH₂-N(L₅)-，-O-CH₂-O-，-O-CH(L₃)-O，-S-CH(L₃)-O，-O-L₃-，-N-L₃-，-S-L₃-，-N(L₃)-L₅-and L₃；

Each A₁-A_mL of_4B，L_4CAnd L_4NAre independently selected from C ═ O, C ═ S,a set of compositions;

L₃and L₅As previously defined;

each A₁-A_mZ on each carbon atom of_A-1Each A₁-A_mZ of (A)_A-2，Z_NT，Z_CT-1And Z_CT-2Each independently selected from the group consisting of: h, CH₃，C₂H₅，C₃H₇，CF₃，C₂F₅，C₃F₇Substituted and unsubstituted hydrocarbon groups, substituted and unsubstituted perfluoroalkyl groups, and substituted and unsubstituted halogenated hydrocarbon groups;

each A₁-A_mR on each carbon atom of_AEach A₁-A_mR on each carbon atom of_B，R_NTAnd R_CTEach independently selected from the group consisting of: substituted and unsubstituted imidazolyl, substituted and unsubstituted quaternidino, substituted and unsubstituted carboxy, substituted and unsubstituted carboxamido, substituted and unsubstituted hydrocarbyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocycloalkyl, substituted and unsubstituted hydrocarbyloxy, substituted and unsubstituted hydrocarbylthio, substituted and unsubstituted hydrocarbylamino, substituted and unsubstituted hydrocarbonyl, substituted and unsubstituted perfluoroalkyl, substituted and unsubstituted halocarbyl, substituted and unsubstituted aryl, and substituted and unsubstituted heteroaryl;

when a is one₁-A_mR on each carbon atom when p is an integer of not less than 2_AOr R_BZ, which may be identical or different, on each carbon atom_A-1May be the same or different;

thiols can further form disulfide bridges.

Examples of Polypeptides HPP/HPC

In certain embodiments, the polypeptide HPP/HPC comprises a structure having a formula selected from the group consisting of: the structural formula 2, the structural formula 3, the structural formula 4, the structural formula 5, the structural formula 6, the structural formula 7, the structural formula 8, the structural formula 9, the structural formula 10, the structural formula 11, the structural formula 12, the structural formula 13, the structural formula 14, the structural formula 15, the structural formula 16, the structural formula 17, the structural formula 18, the structural formula 19, the structural formula 20, the structural formula 21, the structural formula 22, the structural formula 23, the structural formula 24, the structural formula 25, the structural formula 26, the structural formula 27, the structural formula 28, the structural formula 29, the structural formula 30, the structural formula 31, the structural formula 32, the structural formula 33, the structural formula 34, the structural

R is selected from H, substituted and unsubstituted alkyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocycloalkyl, substituted and unsubstituted alkoxy, substituted and unsubstituted alkylthio, substituted and unsubstituted alkylamino, substituted and unsubstituted aryl, substituted and unsubstituted heteroaryl;

X，X₄，X₅，X₆，X₇，X₈，X₉，X₁₀，X₂₁，X₂₂，X₂₃，X₂₄，X₂₅，X₂₆and X₂₇Each independently selected from C ═ O, COO and CH₂OCO，CSO，CH₂OCO，COOCH₂OCO，COCH₂OCO，CH₂-O-CH(CH₂OR₄)₂，CH₂-O-CH(CH₂OCOR₄)₂，SO₂，PO(OR)，NO₂，NO，O，S，NR₅And none;

R₁，R₂，R₄，R₅，R₆，R₇，R₈，R₉，R₁₀，R₂₁，R₂₂，R₂₃，R₂₄，R₂₅，R₂₆and R₂₇Each independently selected from H, O, NO₂Substituted and unsubstituted alkyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocycloalkyl, substituted and unsubstituted alkoxy, substituted and unsubstituted alkylthio, substituted and unsubstituted alkylamino, substituted and unsubstituted alkenyl, substituted and unsubstituted alkynyl, substituted and unsubstituted aryl, and substituted and unsubstituted heteroaryl;

unless otherwise noted, the term "HA" as used herein means no acid or pharmaceutically acceptable acid, such as hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, sulfuric acid, sulfurous acid, phosphoric acid, phosphorous acid, phosphonic acid, isonicotinic acid, acetic acid, lactic acid, salicylic acid, citric acid, tartaric acid, pantothenic acid, acid tartaric acid, ascorbic acid, succinic acid, maleic acid, gentisic acid (gentisic acid), fumaric acid, gluconic acid, glucuronic acid (glucuronic acid), saccharic acid, formic acid, benzoic acid, glutamic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid or pamoic acid; and

ar is selected from the group consisting of: phenyl, 2' -naphthyl, 4-iodophenyl, substituted and unsubstituted aryl, and substituted and unsubstituted heteroaryl.

The corresponding parent drugs for the polypeptide HPP/HPC having the structure represented by structural formulae 2-345 are set forth in tables A (I) and (II):

TABLE A parent peptide (I) of the polypeptide HPP/HPC having the structure represented by structural formulae 2-345

(II)

[0073] The structures of unusual amino acids appearing in SEQ ID nos. 1 to 172 (Xaa1, Xaa2, Xaa3, Xaa4, Xaa5, Xaa7, Xaa9, Xaa13, Xaa14, Xaa15, Xaa17, Xaa19, Xaa21, Xaa22, Xaa23, Xaa24, Xaa25, and Xaa26) are listed in table B:

unusual amino acids found in SEQ ID Nos. 1-172

In certain examples, the polypeptide HPP/HPC has a structure selected from the group consisting of compounds represented by structural formulae 2-345 as previously defined, including stereoisomers and pharmaceutically acceptable salts thereof, wherein;

HA，Ar，X，X₄，X₅，X₆，X₇，X₈，X₉，X₁₀，X₂₁，X₂₂，X₂₃，X₂₄，X₂₅，X₂₆and X₂₇As previously defined;

r is selected from the group consisting of H, substituted and unsubstituted hydrocarbyl of 1 to 20 carbon atoms, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocycloalkenyl, substituted and unsubstituted hydrocarbyloxy of 1 to 20 carbon atoms, substituted and unsubstituted hydrocarbylthio of 1 to 20 carbon atoms, substituted and unsubstituted hydrocarbylamino of 1 to 20 carbon atoms, substituted and unsubstituted aryl, and substituted and unsubstituted heteroaryl;

R₁，R₂，R₄，R₅，R₆，R₇，R₈，R₉，R₁₀，R₂₁，R₂₂，R₂₃，R₂₄，R₂₅，R₂₆and R₂₇Each independently selected from the group consisting of H, O, substituted and unsubstituted hydrocarbyl of 1 to 20 carbon atoms, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocycloalkenyl, substituted and unsubstituted hydrocarbyloxy of 1 to 20 carbon atoms, substituted and unsubstituted hydrocarbylthio of 1 to 20 carbon atoms, substituted and unsubstituted hydrocarbylamino of 1 to 20 carbon atoms, substituted and unsubstituted alkenyl of 1 to 20 carbon atoms, substituted and unsubstituted alkynyl of 1 to 20 carbon atoms, substituted and unsubstituted aryl, and substituted and unsubstituted heteroaryl;

in some examples, the polypeptide HPP/HPC comprises a structure selected from the group consisting of formula 1a, formula 1b, formula 1c, formula 1d, formula 1e, formula 1f, formula 1g, and formula 1 h:

structural formula 1a

Structural formula 1b

Structural formula 1c

Structural formula 1d

Structural formula 1e

Structural formula 1f

Structural formula 1g

Structural formula 1h

Including stereoisomers and pharmaceutically acceptable salts thereof, wherein:

X₄，X₅，X₆，X₇，X₈，X₉，R₄，R₅，R₆，R₇，R₈，R₉and HA is as previously defined.

Solid phase synthesis of polypeptide HPP/HPC

Shorter polypeptides (< 10 amino acids) can be synthesized by liquid phase, but longer polypeptides are difficult to synthesize by liquid phase. The polypeptide HPP/HPC disclosed herein is a modified polypeptide that cannot be synthesized (from C-terminus to N-terminus) by standard solid phase synthesis methods.

In certain examples, the polypeptide HPP/HPC is synthesized from N-terminus to C-terminus, wherein the method of synthesis comprises the steps of:

the method for preparing polypeptide HPC by solid phase synthesis comprises:

a) providing chemically modified resins (such as triphenylchloride resins and carbonate resins);

b) protection of the carboxyl groups of all natural or modified amino acid fragments with protecting groups such as 2- (4-nitrophenylsulfonyl) ethyl and 9-fluorenylmethyl according to polypeptide HPC to form a COOH-protected amino acid fragment;

c) attaching the N-terminal amino acid residue of the COOH-protected polypeptide HPC to a chemically modified resin to form an immobilized COOH-protected polypeptide HPC precursor having one amino acid residue;

d) deprotecting the immobilized COOH-protected polypeptide HPC precursor with a deprotection reagent (e.g., 1% DBU/20% pyridine/79% DMF, or other reagent capable of deprotecting the protected carboxyl group); obtaining an immobilized COOH-unprotected polypeptide HPC precursor having an amino acid residue;

e) repeating steps c) and c) with a COOH-protected amino acid fragment of the polypeptide HPC

d) Until an immobilized C-unprotected precursor of the polypeptide HPC is formed having all but the C-terminal amino acid residue of the amino acid fragment;

f) linking the C-terminal amino acid residue and R by covalent linkage_TTo form a modified C-terminal amino acid, wherein R_TA transport unit selected from the group consisting of substituted and unsubstituted hydrocarbyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocycloalkyl, substituted and unsubstituted hydrocarbyloxy, substituted and unsubstituted alkenyl, substituted and unsubstituted alkynyl, substituted and unsubstituted aryl, substituted and unsubstituted heteroaryl, and from the group consisting of formula Na, formula Nb, formula Nc, formula Nd, formula Ne, formula Nf, formula Ng, formula Nh, formula Ni, formula Nj, formula Nk, formula Nl, formula Nm, formula Nn, formula No, formula Np, formula Nq, and formula Nr as previously defined;

g) attaching the modified C-terminal amino acid to the immobilized C-unprotected polypeptide HPC precursor from step e) to form an immobilized polypeptide HPC; and

h) the immobilized polypeptide HPC is released from the resin to yield the polypeptide HPC.

In certain examples, the chemically modified resin may be directly linked to the protected amino acid, or the chemically modified resin may be first further modified and then linked to the COOH-protected amino acid;

in certain embodiments, the polypeptide HPP/HPC comprising a transport unit at the C-terminal amino acid is prepared by linking a C-terminal amino acid to a polypeptide chain, wherein the carboxyl group is linked to a transport unit as defined above. For example, the transport unit is selected from the group consisting of formula Na, formula Nb, formula Nc, formula Nd, formula Ne, formula Nf, formula Ng, formula Nh, formula Ni, formula Nj, formula Nk, formula Nl, formula Nm, formula Nn, formula No, formula Np, formula Nq, and formula Nr as previously defined.

In certain embodiments, the polypeptide HPP/HPC having a transport unit at the C-terminal amino acid is prepared by linking the C-terminal amino acid to a polypeptide chain, wherein the carboxyl group of the C-terminal amino acid is protected, then deprotected, and finally linked to a transport unit. For example, the transport unit is selected from the group consisting of formula Na, formula Nb, formula Nc, formula Nd, formula Ne, formula Nf, formula Ng, formula Nh, formula Ni, formula Nj, formula Nk, formula Nl, formula Nm, formula Nn, formula No, formula Np, formula Nq, and formula Nr as previously defined.

In certain embodiments, the yield of the released polypeptide HPP/HPC is high. For example, in certain embodiments, the resin is a triphenylchloride resin wherein the liberating step can be accomplished by reaction with TFA/DCM (e.g., 5%). In certain embodiments, the resin is a carbonate resin and the liberating step can be accomplished by reaction with Pd — C (e.g., a 10% solution of methanol) and hydrogen.

In certain embodiments, the coupling reaction is carried out in the presence of a coupling agent. Examples of coupling agents include, but are not limited to, HBTU/DIPEA/HOBt, TBTU/DIPEA/HOBt, BOP/DIPEA/HOBt, HATU/DIPEA/HOBt, and DIC/HOB, and combinations thereof.

Unless otherwise noted, herein:

"DBU" means 1, 8-diazabicyclo [5, 4, 0] undec-7-ene;

"DMF" refers to dimethylformamide;

"DIPEA" refers to N, N-diisopropylethylamine;

"HBTU" means O-benzotriazole-N, N, N ', N' -tetramethyl-urea-hexafluoro-phosphate;

"HOBt" means 1-hydroxybenzotriazole;

"TBTU" refers to O-benzotriazole-N, N, N ', N' -tetramethyl-urea-tetrafluoroborate;

"BOP" means benzotriazol-1-yl-N-oxy-tris (pyrrolidine) phosphonium hexafluorophosphate salt;

"HATU" refers to N- [ (dimethylamino) -1H-1, 2, 3-triazole [4, 5-b ] -pyrido-1-ylmethylene ] -N-methylmethanaminium hexafluorophosphate N-oxide salt;

"DIC" refers to diisopropylcarbodiimide; and

"TFA" refers to trifluoroacetic acid.

Pharmaceutical compositions containing HPP/HPC

Another aspect of the invention relates to a pharmaceutical composition comprising at least one polypeptide HPP/HPC and a pharmaceutically acceptable carrier.

The term "pharmaceutically acceptable carrier" as used herein refers to a pharmaceutically acceptable material, composition or medium, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material, involved in carrying or transporting HPP/HPC from one location, body fluid, tissue, organ (internal or external), or part of the body to another location, body fluid, tissue, organ (internal or external), or part of the body.

Each carrier is "pharmaceutically acceptable" in that it is compatible with the other ingredients of the formulation (e.g., HPP/HPC), and is suitable for use in contact with the tissues or organs of a biological system without undue toxicity, irritation, allergic response, immunogenicity, or other problems or complications commensurate with a reasonable benefit/risk ratio.

Examples of materials that may serve as pharmaceutically acceptable carriers include: 1) sugars such as lactose, glucose and sucrose; 2) starches, such as corn starch and potato starch; 3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; 4) powdered gum tragacanth; 5) malt; 6) gelatin; 7) talc powder; 8) adjuvants, such as cocoa butter and suppository waxes; 9) oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; 10) glycols, such as propylene glycol; 11) polyhydroxy compounds such as glycerol, sorbitol, mannitol and polyethylene glycol; 12) esters, such as ethyl oleate and ethyl laurate; 13) agar; 14) buffering agents such as magnesium hydroxide and aluminum hydroxide; 15) alginic acid; 16) pyrogen-free water; 17) isotonic saline; 18) ringer's solution; 19) alcohols such as ethanol and propanol; 20) a phosphate buffer solution; and 21) other non-toxic compatible substances such as acetone for use in pharmaceutical formulations.

The pharmaceutical compositions may contain pharmaceutically acceptable auxiliary substances as required to achieve near physiological conditions such as pH adjusting and buffering agents, toxicity adjusting agents and the like, for example, sodium acetate, sodium chloride, potassium chloride, calcium chloride, sodium lactate and the like.

In one embodiment, the pharmaceutically acceptable carrier is an aqueous carrier, such as a buffered saline solution and the like. In certain embodiments, the pharmaceutically acceptable carrier is a polar solvent, such as water, acetone, and alcohols.

The concentration of HPP/HPC in these formulations can vary over a wide range and can be selected based primarily on the volume, viscosity, body weight and the like of the fluid, depending on the particular mode of administration selected and the requirements of the biological system. For example, the concentration (by weight) may be 0.0001% to 100%, 0.001% to 50%, 0.01% to 30%, 0.1% to 10%.

The compositions of the present invention may be administered for prophylactic, therapeutic, and/or hygienic uses. Such administration may be by topical, mucosal, such as buccal, nasal, vaginal, rectal, injection, transdermal, subcutaneous, intramuscular, intravenous, inhalation, ocular, and other suitable routes. The pharmaceutical compositions can be administered in a variety of unit dosage forms depending on the method of administration. For example, unit dosage forms suitable for oral administration include powders, tablets, pills, capsules, and lozenges.

Thus, a typical pharmaceutical composition for intravenous administration may be about 10 days per subject^-10Grams to about 100 grams, about 10^-10To about 10^-3Gram, about 10^-9Gram to about 10^-6Gram, about 10^-6From about 0.001 to about 100 grams, from about 0.01 to about 10 grams, or from about 0.01 to about 1 gram. Each timeThe daily dosage for each subject may range from about 0.01 mg to about 50 g. The actual methods for preparing compositions for injection administration are well known in the art or will be apparent to those skilled in the art, and are described in more detail in publications such as Remington's Pharmaceutical Science, 15th ed., Mack Publishing Company, Easton, Pa. (1980).

Use of HPP/HPC

i) A method of penetrating a biological barrier.

Another aspect of the invention relates to a method of penetrating one or more biological barriers in a biological subject using a composition of the invention. The method comprises the step of administering to the biological subject an HPP/HPC or a polypeptide or polypeptide-related compound, or a pharmaceutical composition thereof. In certain embodiments, HPP/HPC exhibits a penetration rate of one or more biological barriers that is greater than about 10-fold or greater, 50-fold or greater, greater than about 100-fold or greater, greater than about 200-fold or greater, greater than about 300-fold or greater, greater than about 500-fold or greater, greater than about 1000-fold or greater than its parent drug.

The term "biological barrier" refers herein to a biological layer that separates an environment into distinct spatial regions or compartments (compositions) that regulates (e.g., restricts, limits, enhances, or does not act upon) the passage, penetration, or transport of a substance (substance) or substance (matter) from one compartment/region to another. The different spatial regions or compartments referred to herein may have the same or different chemical or biological environments. Reference herein to a biological layer includes, but is not limited to, a biological membrane, a cellular layer, a biological structure, an object, an organism, an inner surface of an organ or a body cavity, an outer surface of an object, an organism, an organ or a body cavity, or any combination or plurality thereof.

Examples of biological membranes include lipid bilayer structures, eukaryotic cell membranes, prokaryotic cell membranes, and intracellular membranes (e.g., nuclear or organelle membranes such as golgi membrane or envelope, rough and smooth endoplasmic reticulum membrane or envelope, ribosomal membrane or envelope, vacuolar membrane or envelope, vesicle membrane or envelope, liposome membrane or envelope, mitochondrial membrane or envelope, lysosomal membrane or envelope, nuclear membrane or envelope, chloroplast membrane or envelope, plastid (plastic) membrane or envelope, peroxisomal membrane or envelope, or microbody membrane or envelope).

The lipid bilayer structure of the present invention is a bilayer structure of lipid molecules, including, but not limited to, phospholipids and cholesterol. In a particular embodiment, the bilayer structured lipid is an amphiphilic molecule consisting of a polar end group and a non-polar fatty acid tail chain. The bilayer structure is composed of two lipid layers, and the hydrocarbon tail chains of the lipid layers are arranged to face each other and gather together to form an oily core through a hydrophobic effect, and the charged end groups of the lipid layers face aqueous phase solutions on two sides of the membrane. In another particular embodiment, the lipid bilayer structure may contain one or more protein and/or carbohydrate molecules embedded therein.

Examples of cell layers include eukaryotic cell layers (e.g., epithelium, lamina propria (lamina propria), and smooth muscle or mucosal muscularis (in the gastrointestinal tract)), prokaryotic cell layers (e.g., surface layer or S layer, which refers to a two-dimensional structural monolayer composed of the same proteins or glycoproteins, in particular, S layer refers to a portion of the cell envelope that is commonly present in bacteria and archaea (archaea)), biofilms (a structured microbial community encapsulated in a spontaneously-forming polymer matrix that adheres to active or inert surfaces), and plant cell layers (e.g., epidermis). The cell can be a normal cell or a diseased cell (e.g., a disease cell, a cancer cell).

Examples of biological structures include structures sealed by tight or closed junctions, which provide a barrier to the entry of toxins, bacteria and viruses, such as the blood milk barrier and the Blood Brain Barrier (BBB). In particular, the BBB consists of a class of impermeable endothelium that forms a physical barrier in conjunction with adjacent endothelial cells through tight junctions and a transport barrier containing efflux transport pumps. Biological structures may also include mixtures of cells, proteins, and carbohydrates (e.g., thrombi).

Examples of the inner surface of the subject, organism, organ or body cavity include buccal mucosa (buccal mucosa), esophageal mucosa, gastric mucosa, intestinal mucosa, olfactory mucosa, oral mucosa, bronchial mucosa, uterine mucosa and endometrium (mucosa of uterus, cell wall lining of pollen grains, or lining wall of spores), or a combination or plura thereof.

Examples of the external surface of a subject, organism, organ or body cavity include capillaries (e.g., capillaries of heart tissue), the membrane of the mucosa associated with the skin (e.g., in the nostrils, lips, ears, reproductive area, and anus), the external surface of an organ (e.g., liver, lung, stomach, brain, kidney, heart, ears, eyes, nose, mouth, tongue, colon, pancreas, gall bladder, duodenum, rectum, stomach, large intestine (colonrectum), intestine, vein, respiratory system, blood vessels, anorectum, and anal itch), the skin, the stratum corneum (e.g., dead cell layer of epithelial cells or keratinocytes or the superficial layer of overlapping cells overlying hair shafts of animals (superficial layer), the multi-layered structure outside the epidermis of various invertebrates, the stratum corneum or polymer cutin (cutin) and/or the glue film (cutin)), the outer layer of cell walls of pollen grains, or the outer wall of spores, and combinations or pluralities thereof.

In addition, the biological barrier further comprises a carbohydrate layer, a protein layer or any other biological layer, or a combination or plurality thereof. For example, skin is a biological barrier with multiple biological layers. The skin contains an epithelial layer (outer surface), a cortex layer and a hypodermal layer. The epithelial layer contains a multi-layered structure including a basal cell layer, a spinocellular layer, a granular cell layer, and a horny layer. Cells in the epithelium are called keratinocytes. The stratum corneum (also called "horny layer") is the outermost layer of the epithelium, where the cells are flat in shape and shaped like scales (scaly). These cells contain a large amount of keratin and are arranged in layers to impart toughness, oil repellency and water repellency to the skin surface.

ii) methods of diagnosing symptoms in biological systems

Another aspect of the invention relates to methods of using the compositions of the invention for diagnosing a condition in a biological system. The method comprises the following steps:

1) administering to a biological subject a composition comprising a polypeptide or a polypeptide-related compound HPP/HPC;

2) detecting the presence, location or amount of the HPP/HPC, the functional unit of the HPP/HPC or a metabolite thereof in the biological subject; and

3) determining a symptom in the biological subject.

In certain embodiments, the HPPs/HPCs (or agents sheared from the HPPs) aggregate at the site of action where symptoms appear. In certain embodiments, the presence, location, or content of functional units of the HPP/HPC is also detected. In certain embodiments, the occurrence, progression, or slowing associated with a symptom (e.g., cancer) is also determined.

In certain embodiments, the HPP/HPC is labeled or conjugated to a detectable substance. Alternatively, HPP/HPC is prepared to contain radioisotopes for detection. A variety of detectable substances are available, roughly classified into the following categories:

a) radioisotopes, e.g.³⁵S、¹⁴C、¹³C、¹⁵N、¹²⁵I、³H. And¹³¹I. the diagnostic substance may be labelled with a radioisotope by techniques well known in the art, and radioactivity may be measured by scintillation counting; in addition, the diagnostic substance can detect carbon and nitrogen labels by electron paramagnetic resonance with spin labeling.

b) Fluorescent substances such as BODIPY, BODIPY analogs, rare earth chelates (europium chelates), fluorescein and its derivatives, FITC, 5, 6 carboxyfluorescein, rhodamine and its derivatives, dansyl, Lissamine, phycoerythrin, green fluorescent protein, yellow fluorescent protein, Red fluorescent protein, and Texas Red (Texas Red). Fluorescence can be quantified by a fluorometer.

c) Various enzyme-substrate substances, such as luciferases (e.g., firefly luciferase and bacterial luciferases), luciferin, 2, 3-dihydrophthalazinediones (2, 3-dihydrophthalazinediones), malate dehydrogenase, urease, peroxidases such as horseradish peroxidase (HRPO), alkaline phosphatase, β -galactosidase, glucoamylase, lysozyme, polysaccharide oxidases (e.g., glucose oxidase, galactose oxidase and glucose-6-phosphate dehydrogenase), heterocyclic oxidases (e.g., uricase and xanthine oxidase), lactose peroxidase, microperoxidase, and the like. Enzyme-substrate combinations include, for example: (i) horseradish peroxidase (HRPO) with catalase as a substrate, wherein the catalase oxidizes a dye precursor (e.g., o-phenylenediamine (OPD) or 3,3 ', 5, 5' -tetramethylbenzidine hydrochloride (TMB), Alkaline Phosphatase (AP) with p-nitrophenyl phosphate as a chromogenic substrate, and (iii) β -D-galactosidase (. beta. -D-Gal) with either a chromogenic substrate (e.g., p-nitrophenyl-. beta. -D-galactosidase) or a fluorogenic substrate 4-methylumbelliferyl-. beta. -D-galactosidase).

In certain embodiments, the detectable substance need not be conjugated to the diagnostic substance, but the presence of the diagnostic substance can be identified and the diagnostic substance can be detected.

In certain embodiments, the HPP/HPC of a polypeptide may be provided by way of a kit (kit), i.e., a predetermined set of reagents packaged together with instructions for conducting a diagnostic assay. When the HPP/HPC is labeled with an enzyme, the kit may include a substrate and cofactors required for the enzyme (e.g., a substrate precursor that provides a detectable chromophore or fluorophore). In addition, other additives such as stabilizers, buffers (e.g., blocking buffers or lysis buffers), and the like may also be included. The relative amounts of the various reagents may be varied over a wide range to provide various concentrations of the reagent solutions which substantially significantly optimize the sensitivity of the assay. In particular, these reagents may be provided in the form of a dry powder (usually freeze-dried) comprising adjuvants which, once dissolved, provide a solution of the reagents in the appropriate concentration.

iii) methods for screening substances with desired characteristics (desired characteristics)

Another aspect of the invention relates to a method for screening HPPs/HPCs for desirable characteristics.

In certain embodiments, the method comprises:

1) covalently linking a test functional unit to a transport unit via a linker to form a test composition (or covalently linking a functional unit to a test transport unit via a linker, or covalently linking a functional unit to a transport unit via a test linker)

2) Administering the test composition to a biological system; and

3) determining whether the test composition has the desired property or characteristic.

In one embodiment, the desired characteristics may include, for example, 1) the ability of the test functional unit to form a high penetration composition or its conversion back to the parent drug; 2) testing the penetration capacity and/or speed of the composition; 3) testing the efficiency and/or effectiveness of the composition; 4) testing the transport capacity of the transport unit; and 5) testing the shearability of the linker

iv) methods for treating a condition in a biological subject

Another aspect of the invention relates to a method of using the composition of the invention for treating a condition of a biological system. The method comprises administering the pharmaceutical composition to the biological system.

The term "treating" as used herein refers to curing, alleviating, inhibiting, or preventing. The term "treating" or "treatment" as used herein refers to curing, alleviating, inhibiting, or preventing. The term "treat" as used herein refers to cure, alleviate, inhibit, or prevent.

The terms "biological system," "biological subject," or "subject" as used herein refer to an organ, a group of organs, organisms, or a group of organisms that work together to accomplish a particular task. The term "organism" refers herein to a collection of molecules that function more or less as a stable whole and have living properties, such as animals, plants, fungi, or microorganisms.

The term "animal" as used herein refers to a eukaryotic organism characterized by active activity. Examples of animals include, but are not limited to, vertebrates (e.g., humans, mammals, birds, reptiles, amphibians, fish, gills, and leptospira), tunicates (e.g., gobies, urothecas, deepwater ecthyma, ascidioides), somites (e.g., entomophytes, polypodas, malaapada, arachnids, acropoda, crustaceans, and annelids), athymia (antarea), and worms (e.g., rotifers).

The term "plant" as used herein refers to an organism belonging to the kingdom Plantae. Examples of plants include, but are not limited to, spermatophytes, bryophytes, ferns, and pteridopsis. Examples of seed plants include, but are not limited to, cycads (cycads), ginsengs (ginkgo), conifers (conifers), gnetophylls (gnetophylls), angiosperms. Examples of bryophytes include, but are not limited to, lichen (liverworts), hornworts, and mosses. Examples of ferns include, but are not limited to, the order chrysosporium (ophthalles) (e.g., paradoxus-tongues, agriophyta), the family Synechocystidae, and phyllanthus gracilis (leptosporangium ferns). Examples of pseudopteridophytes include, but are not limited to, the class of the pinopidae (e.g., lycopodium, selaginella (spikemoses), and leek (quillworts)), the family of the conidioidea (e.g., phylum lycopodium and order conidioidea), and the family of the equisetum (e.g., class equisetum).

The term "fungus" as used herein refers to a eukaryotic organism that is a member of the kingdom fungi. Examples of fungi include, but are not limited to, Chytridiomycota, Blastocladiomycota, Neocallimastigomycota, zygomycota, Synechomycota, Ascomycota, and Basidiomycota.

The term "microorganism" as used herein refers to microscopic organisms (e.g., on the order of microns in length). Examples of microorganisms include, but are not limited to, bacteria, fungi, archaea, protozoa, and microscopic plants (e.g., green algae) and microscopic animals (e.g., plankton, trichuris, and amoebas).

Some examples of conditions treatable by the present methods include conditions treatable by prodrugs of HPP/HPC.

v) Polypeptides and Polypeptides HPP/HPC and methods of use of pharmaceutical compositions thereof in therapy

Another aspect of the invention relates to a method of treating a condition in a biological system or a subject using a HPP/HPC of a polypeptide, or a pharmaceutical composition thereof, by administering the polypeptide HPP/HPC, or a pharmaceutical composition thereof, to the biological system or the subject.

Polypeptides and polypeptide-related compounds are useful for the broad regulation of a variety of biological processes in biological systems. Symptoms associated with these biological processes can be treated by the corresponding polypeptide or polypeptide-related compound, and thus can also be treated with the polypeptide HPP or HPC, and pharmaceutical compositions thereof.

These symptoms include, but are not limited to: aging, angina, antithrombin deficiency, arrhythmia, atherosclerosis, atrial fibrillation, atrial flutter, thrombosis, cardiac ischemia, cardiac surgery, cardiomyopathy, cardiovascular abnormalities, carotid artery disease, chest pain, circulatory disorders, claudication, collagen vascular disease, congenital heart disease, congestive heart failure, coronary heart disease, diabetes and hypertension, hyperlipidemia, dysrhythmia, elevated triglycerides, heart defects, heart disease, heart failure, valvular heart disease, hemangioma, high cholesterol, hypertriglyceridemia, intermittent claudication, hypertension, kawasaki disease, heart attack, myocardial ischemia, postural hypotension, peripheral arterial disease, peripheral arterial occlusive disease, peripheral vascular disease, Raynaud's disease, smoking cessation, tachycardia, thrombosis, lower limb varicose veins, vascular disease, venous ulcer, gingivitis, angina, cardiac ischemia, cardiac arrhythmia, cardiac insufficiency, cardiac, Gum disease, halitosis, oral cancer, periodontal disease, temporomandibular joint disorder, temporomandibular joint syndrome, sunburn, acne, skin aging, alopecia, anemia, athlete's foot, atopic dermatitis, bedsore (decubitus ulcer), bunion, burn infection, herpes labialis (herpes labialis infection), congenital skin disease, contact dermatitis, cutaneous lupus erythematosus, diabetic foot ulcer, eczema, excessive sweating, Fabry's disease, fungal infection, genital herpes, genital warts, hair loss, hand dermatitis, head louse, hemangioma, hereditary angioedema, herpes simplex infection, herpes zoster neuralgia, urticaria, ichthyosis, ischemic foot ulcer, keratosis, lupus, male pattern baldness, malignant melanoma, benign melanoma, molluscum contagiosum, mycosis fungoides, onychomycosis, herpes vulgaris, herpes zoster, psoriasis, tinea pedis, atopic dermatitis, skin ulcer, skin, Postherpetic neuralgia, pressure sores, psoriasis and psoriasis, psoriatic arthritis, razor jolt, acne rosacea, sarcoidosis, scalp disorders, scar tissue, scleroderma, seborrhea, seborrheic dermatitis, herpes zoster, skin cancer, skin infections, skin lipomas, skin wounds, freckles, sporotrichosis, staphylococcal skin infections, blood stasis dermatitis, stretch marks, systemic fungal infections, sun-toxicity, tinea capitis, tinea versicolor, urticaria, vitiligo, warts, wounds, acromegaly, adrenal cancer, congenital adrenal cortical hyperplasia, diabetes mellitus (types I and II), diabetes mellitus (type I), diabetes mellitus (type II), diabetic gastroparesis, diabetic kidney disorders, diabetic macular edema, diabetic neuropathy, diabetic retinopathy, diabetic vitreorrhagia, Dyslipidemia, female hormone deficiency/abnormality, fredrickson type III hyperlipidemia, growth hormone deficiency/abnormality, gynecomastia, alopecia, hyperlipidemia, hormone deficiency, hot flashes, hyperparathyroidism, idiopathic short stature, indications: type II diabetes, male hormone deficiency/abnormality, McCune-Albright syndrome (multiple fiber dystrophy), menopausal disorders, metabolic syndrome, obesity, ovarian cancer, pancreatic disease, pancreatitis, parathyroid cancer, parathyroid disease, parathyroid disorder (parathyroid disorders), peri-menopause, pituitary disease, polycystic ovary syndrome, post-menopause disease, post-menopause osteoporosis, precocious puberty, small hypersecretion of insulin, severe stature dwarfism, sexual dysfunction, thyroid disease, thyroid disorder, Telner's syndrome, wilson's disease, abdominal cancer, cardiac achalasia, alpha 1 antitrypsin deficiency, anal fissure, appendicitis, Barrett's esophagus, biliary tract cancer, intestinal tract dysfunction, abdominal cavity disease, chronic diarrhea, Clostridium difficile-associated diarrhea, colon cancer, pancreatic, Colonic polyps, large intestine cancer, constipation, crohn's disease, diabetic gastroparesis, tumors of the digestive system, duodenal ulcers, fabry's disease, fecal incontinence, functional dyspepsia, gallbladder diseases, stomach cancer, gastric ulcers, gastroenteritis, gastroesophageal reflux disease, gastrointestinal diseases and disorders, gastroparesis, heartburn, helicobacter pylori, hemorrhoids, hepatic encephalopathy, hepatitis, intestinal obstruction, infectious enteritis, inflammatory bowel disease, intra-abdominal infection, irritable bowel syndrome, liver diseases, liver disorders, non-erosive reflux disease, non-ulcerative dyspepsia, organ transplantation for organ rejection, postoperative nausea and vomiting, rectal cancer, rectal disorders, recurrent diarrhea, stomach cancer, stomach discomfort, ulcerative colitis, abnormal blood vessels, acute myeloid leukemia, anemia (non-hodgkin's lymphoma), non-small cell lung cancer, Anemia of cancer, aneurysm, antiphospholipid antibody syndrome, antithrombin deficiency, aplastic anemia, thrombosis, candidiasis, chronic renal anemia, gaucher's disease, hematologic cancer, hematologic disease, paroxysmal hemoglobinuria, bleeding, hypercalcemia, hypogammaglobulinemia, hyponatremia, idiopathic thrombocytopenic purpura, islet cell carcinoma, leukemia, B cell lymphoma, multiple myeloma, myelodysplastic syndrome, myocardial ischemia, occlusion, platelet deficiency, platelet disorders, erythrocyte abnormalities, renal anemia, Zalciley syndrome, sickle cell disease, cell lymphoma, thalassemia, thrombocytopenia, hemophilia, leukocyte abnormalities, acquired immunodeficiency syndrome (AIDS), AIDS-related diseases, acute rhinitis, allergy, asthma, anal dysplasia, Acquired Immune Deficiency Syndrome (AIDS), AIDS-related diseases, acute rhinitis, allergy, and asthma, Bacterial infections, thrush, celiac disease, cervical hyperplasia, chickenpox, chronic fatigue syndrome, the common cold, common variant immunodeficiency disease, bacterial conjunctivitis, chronic obstructive pulmonary disease, cutaneous candidiasis, cutaneous T-cell lymphoma, cytomegalovirus infection, dermatomyositis, fever, graft-versus-host disease, hepatitis B, hepatitis C, aids virus infection, aids virus/aids, human papilloma virus infection, hypogammaglobulinemia, idiopathic inflammatory myopathy, influenza, intra-abdominal infection, kaposi's sarcoma, lupus, lyme disease tick, mycobacterium avium complex infection, meningitis, onychomycosis, oral candidiasis, pneumonia, polymyositis (inflammatory muscle disease), postherpetic neuralgia (postherpetic neuralgias), primary immunodeficiency, respiratory syncytial virus infection, rheumatic fever, chronic inflammation of the lungs, chronic inflammatory disease, chronic inflammation of the skin, chronic inflammation of, Allergic rhinitis, rotavirus infection, sarcoidosis, sepsis and septicemia, sexually transmitted diseases, herpes zoster, sjogren's syndrome, smallpox, soft tissue infection, staphylococcal skin infection, streptococcal laryngitis, systemic candida, systemic lupus erythematosus, throat and tonsil infections, urticaria, vancomycin-resistant enterococci, west nile virus infection, acromegaly, ankylosing spondylitis, bone loss, sports trauma, bone disease, bone metastasis, breast pain, bunions, bursitis, carpal tunnel syndrome, cartilage injury, chest pain, chronic back pain, chronic low back and leg pain, chronic shoulder pain, claudication, congenital lactic acidosis, connective tissue disease, dermatomyositis, telun contracture, fibromyalgia, frozen shoulder, adhesive capsulitis, gout (hyperuricemia), diabetes mellitus, and other diseases, Idiopathic inflammatory myopathy, intermittent claudication, joint injury, knee injury, multiple sclerosis, muscle pain, muscular dystrophy, musculoskeletal disease, myasthenia gravis, generalized myasthenia gravis, orthopedics, osteoarthritis, osteomyelitis, osteoporosis, osteosarcoma, pekia, medial meniscus resections, parathyroid disease, postmenopausal osteopenia, postmenopausal osteoporosis, reflex sympathetic dystrophy syndrome, rheumatoid arthritis, sciatica, spinal cord disease, spinal cord tumors, spinal cord arthroplasty, sprains, tendon injury, tennis elbow, tic disorders, anal dysplasia, benign prostatic hyperplasia, bladder cancer, bladder disorder, hematological cancer, catheter complications, chronic pelvic pain, diabetic kidney disease, enuresis, erectile dysfunction, fabry's disease, chronic pelvic pain, renal failure, chronic pelvic pain, kidney disease, muscle pain, muscle soreness, muscle wasting, muscle weakness, muscle and muscle weakness, Nocturia, urogenital prolapse, glomerulonephritis, glomerulosclerosis, idiopathic membranous nephropathy, impotence, interstitial cystitis, kidney cancer, kidney disease, renal failure, kidney stones, liver cancer, low testosterone levels, mastectomy, prosthetics, kidney disease, peyronie's disease, premature ejaculation, prostate cancer, prostate disease, prostatic epithelial sarcoma, proteinuria, reiter's syndrome, renal artery disease, renal cell carcinoma, renal failure, testicular cancer, tyrosinemia, urethral stricture, urinary incontinence, urinary tract infection, urothelial cancer, male erectile dysfunction and female sexual dysfunction, systemic blood pressure, artificial abortion, hypotension control, inhibition of platelet coagulation, lung disease, gastrointestinal disease, inflammation, shock, reproductive disease, infertility, obesity, and the like.

Symptoms associated with platelet aggregation include, for example, post-operative, post-carotid endarterectomy thromboembolism, restenosis after coronary angioplasty, thromboembolic syndrome in chronic atrial fibrillation, aortic coronary bypass circulatory system obstruction, myocardial infarction, stroke, multi-embolic dementia, arterial embolic complications in hemodialysis and prosthetic valve patients.

Conditions that may be treated using methods that include the use of the polypeptide HPP/HPC, or pharmaceutical compositions thereof, include, but are not limited to, polypeptide hormone-related conditions, inflammation and related conditions, platelet aggregation-related conditions, neuropeptide-related conditions, microorganism-related conditions, and other conditions modulated by a polypeptide or polypeptide-related compound.

In certain embodiments, the method of treating a condition treatable by a polypeptide comprises administering to a biological system HPP/HPC of a polypeptide or polypeptide-related compound such as angiotensin, angiotensin II antagonist, angiotensin II AT2 receptor, antimicrobial peptide, oxytocin, hormone, antidiuretic hormone, corticotropin, antimicrobial peptide, anti-inflammatory peptide, bradykinin antagonist, endothelin peptide antagonist, gastrin, calcitonin, melanoma-related antigenic peptide, fibronectin peptide, fibrinogen peptide, EAE-inducing peptide, growth factor, growth hormone releasing peptide, somatostatin, hormone releasing hormone, luteinizing hormone releasing hormone, neuropeptide, melanocyte stimulating peptide, sleep inducing peptide, amyloid peptide, deglutition promoting hormone, HPP/HPC of retro-inverso-phagocytosines, enterostatin, melanocortin II, and opioid peptide polypeptides and mimetics

In certain embodiments, a method of treating a polypeptide hormone-related condition comprises administering to a biological system an HPP/HPC of a polypeptide hormone or polypeptide hormone-related compound, or a pharmaceutical composition thereof. In biological systems, hormones regulate a wide range of processes such as energy levels, reproduction, growth and development, homeostasis, and responses to environment, stress, and injury. Polypeptide hormone-related conditions include, but are not limited to

a) In menopause

b) Bone diseases, such as osteoporosis, paget's disease and bone metastases;

c) growth hormone deficiency;

d) hyperthyroidism or hypothyroidism;

e) metabolic diseases, such as obesity, abnormal blood glucose levels, abnormal blood lipid levels, diabetes (type I or/and type II) and complications resulting from diabetes, including diabetic retinopathy, necrotic ulcers and diabetic proteinuria;

f) blood pressure abnormalities, such as hypertension and hypotension;

g) skin conditions such as psoriasis and psoriasis, acne, cystic acne, running sores, abscesses or redness, comedones, papules, pustules, nodules, epidermoid cysts, hair keratinization, vascular skin damage, birthmarks, moles, sagging, scleroderma, vitiligo and related diseases, or age spots (liver spots);

h) autoimmune diseases such as discoid lupus erythematosus, Systemic Lupus Erythematosus (SLE), autoimmune hepatitis, cleroderma, sjogren's syndrome, rheumatoid arthritis, polymyositis, scleroderma, hashimoto's thyroiditis, juvenile diabetes, edison's disease, vitiligo, pernicious anemia, glomerulonephritis, pulmonary fibrosis, Multiple Sclerosis (MS) and crohn's disease;

i) eye diseases such as glaucoma, ocular hypertension, blindness after ophthalmic surgery, cystoid macular edema and impaired vision in warm-blooded animals caused by cataracts;

j) pre-eclamptic toxemia in high risk women;

k) male and female sexual dysfunction;

j) allergies and asthma;

k) insomnia;

l) depression and related diseases;

m) cardiovascular diseases, such as myocardial infarction, unstable angina, peripheral arterial occlusive disease and stroke;

n) tumors, such as benign tumors, breast cancer, colon cancer, oral cancer, lung or other respiratory cancer, skin cancer, uterine cancer, pancreatic cancer, prostate cancer, vaginal cancer, urinary organ cancer, leukemia or other cancers of the blood and lymphoid tissues; and

o) metastases.

In certain embodiments, the method of treating a microbial-related disease comprises administering HPP/HPC of an antimicrobial peptide or antimicrobial peptide-related compound, or a pharmaceutical composition thereof, to a biological system. Examples of microorganism-associated symptoms include, but are not limited to, inflammation and related symptoms

a) Pain;

b) (ii) damage;

c) a microorganism-associated symptom;

d) inflammation-related symptoms, such as prostatitis, prostacystitis, prostatic hypertrophy and fibrosis, hemorrhoids, Kawasaki disease, gastroenteritis, type I membranous glomerulonephritis, Batt's syndrome, chronic uveitis, ankylosing spondylitis, hemophilia arthritis, inflammatory hemorrhoids, radiation proctitis, chronic ulcerative colitis, inflammatory bowel disease, pouchitis, periodontitis, arthritis, and inflammatory diseases of the liver, lung, stomach, brain, kidney, heart, ear, eye, nose, mouth, tongue, colon, pancreas, gall bladder, duodenum, stomach and rectum, colorectal, intestinal, venous, respiratory, vascular, anal and anal itch of the following organs.

In certain embodiments, a method of treating a neuropeptide-related condition comprises administering an HPP/HPC of a neuropeptide or neuropeptide-related compound, or a pharmaceutical composition, to a biological system. Examples of neuropeptide-related conditions include, but are not limited to, pain, and neurodegenerative diseases such as senile dementia and parkinson's disease.

Other symptoms that can be treated by the HPP/HPC or pharmaceutical composition include, but are not limited to, symptoms relating to: symptoms associated with platelet aggregation, such as thromboembolism after surgery, carotid endarterectomy, restenosis after coronary angioplasty, thromboembolic syndrome in chronic atrial fibrillation, aortic coronary bypass circulatory system obstruction, myocardial infarction, stroke, multiembolic dementia, arterial embolic complications in hemodialysis and in patients with prosthetic valves; prenatal, postpartum, anti-AD activity, anti-diuretic activity, brain damage, calcium balance, melanocyte, central nervous system activity and phagocytosis.

In certain embodiments, a method of treating a condition in a subject that is ameliorated or treatable by a polypeptide or polypeptide-related compound comprises administering to the subject a therapeutically effective amount of the polypeptide HPP/HPC, or a pharmaceutical composition.

HPP/HPC or pharmaceutical compositions thereof may be administered to a biological system by any route of administration known in the art, including, but not limited to, oral, intestinal, oral, nasal, topical, rectal, vaginal, aerosol, transmucosal, epithelial, transdermal, dermal, ocular, pulmonary, subcutaneous, and/or injectable routes of administration. The pharmaceutical composition can be administered in a variety of unit dosage forms depending on the mode of administration.

By injection is meant a route of administration commonly associated with injection, including, but not limited to, intravenous, intramuscular, arterial, intrathoracic, intracapsular (intracapsule), intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, sublingual, sub-cuticular, intraarticular (intraarticular), subcapsular, subarachnoid, intraspinal, and/or intrasternal injection and/or instillation.

The HPP/HPC or pharmaceutical composition thereof may be administered to a subject in a formulation or preparation suitable for each route of administration. Formulations useful in the methods of the invention include one or more HPPs/HPCs, one or more pharmaceutically acceptable carriers, and optionally other therapeutic ingredients. The formulations may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy. The active ingredient may be combined with a carrier material to form a single dosage form, the amount of which may vary depending upon the subject being treated and the particular mode of administration. The HPP/HPC may be combined with a carrier material to form a pharmaceutically effective dose, typically in an amount such that a therapeutic effect is produced. Generally, the amount of HPP/HPC may vary from about 1% to about 99% of HPP/HPC, preferably from about 20% to about 70%, based on 100%.

Methods of making these formulations or compositions include the step of associating HPP/HPC with one or more pharmaceutically acceptable carriers and, optionally, one or more accessory ingredients. In general, the formulations can be prepared by uniformly and intimately bringing the HPP/HPC into association with a liquid carrier, a finely divided solid carrier, or both, and then, if necessary, shaping the product.

Dosage forms suitable for oral administration may be capsules, cachets (cachets), pills, tablets, lozenges (using stirred base ingredients, usually sucrose, acacia or tragacanth), powders, granules, or solutions or suspensions in aqueous or non-aqueous liquids, or oil-in-water or water-in-oil liquid emulsions, or elixirs (elixir) or syrups, or lozenges (using an inert base such as gelatin and glycerol, or sucrose and acacia) and/or mouthwashes and the like, each containing a predetermined amount of HPP/HPC as the active ingredient. The compounds may also be administered by bolus (bolus), electuary (electuary), or paste.

In solid dosage forms for oral administration (e.g., capsules, tablets, pills, dragees, powders, granules and the like), HPP/HPC is mixed with one or more pharmaceutically acceptable carriers, such as sodium citrate or dicalcium phosphate, and/or optionally from: (1) fillers or additives, such as starch, lactose, sucrose, glucose, mannitol, and/or silicic acid; (2) binders, such as, for example, hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and/or acacia; (3) humectants, such as glycerol; (4) disintegrants, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; (5) dissolution retarding agents (dissolution reducing agents), such as paraffin; (6) absorption promoters, such as quaternary ammonium compounds; (7) wetting agents, such as acetyl alcohol (acetyl alcohol) and glyceryl monostearate; (8) absorbents such as kaolin and bentonite clay; (9) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and combinations thereof; and (10) a colorant. In the case of capsules, tablets, and pills, the pharmaceutical compositions may also include buffering agents. Solid compositions of a similar type may also be employed as fillers in soft-filled and hard-filled gelatin capsules using, for example, lactose or milk sugar (milk sugars) as well as high molecular weight polyethylene glycols and the like as excipients.

Tablets may be prepared by compression or molding, optionally with the addition of one or more accessory ingredients. Compressed tablets may be prepared by binding agents (for example, gelatin or hydroxypropyl cellulose), lubricating agents, inert diluents, preservatives, disintegrating agents (for example, sodium starch glycolate or cross-linked sodium carboxymethyl cellulose), surface active agents or dispersing agents. Compression molded tablets may be made by compression molding in a suitable machine a powdered beta-lactam antibiotic or a peptidomimetic composition moistened with an inert liquid diluent. Tablets, or other solid dosage forms such as dragees, capsules, pills and granules, optionally scored or otherwise prepared with coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical formulating art. They may also provide sustained or controlled release of HPP/HPC by formulation, e.g., using hydroxypropylmethylcellulose in various ratios to provide a desired release profile, using other polymer matrices, using liposomes, and/or using microspheres. They may be sterilized, for example, by filtration through a bacterial-retaining filter, or by incorporating sterilizing agents into sterile solid compositions which may be dissolved in sterile water, or other sterile injectable vehicles, immediately prior to use. These compositions may optionally also contain soothing agents (pacifying agents) which may release HPP/HPC only, or preferentially, at a particular site in the gastrointestinal tract, optionally by delayed release. Examples of useful embedding compositions include polymeric substances and waxes. HPP/HPC may also be in the form of microcapsules, where appropriate with one or more of the excipients mentioned above.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to HPP/HPC, the liquid dosage forms may contain inert diluents commonly used in the art, such as, for example, water or other solvents, solubilizing agents and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate (ethyl lcarbonate), ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1, 3-butylene glycol, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol (tetrahydrofurfuryl alcohol), polyethylene glycols and fatty acids of sorbitan, and combinations thereof. In addition to inert diluents, oral compositions can also contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming (perfuming) agents, and preserving agents.

Suspensions, in addition to HPP/HPC, may contain suspending agents, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitol esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar, and tragacanth, and combinations thereof.

Formulations for rectal or vaginal administration may be presented as a suppository, which may be prepared by mixing one or more HPPs/HPCs with one or more suitable non-irritating excipients or carriers comprising, for example, cocoa butter, polyethylene glycol, a suppository wax or salicylate, which is solid at room temperature but liquid at body temperature and therefore will melt in the rectum or vaginal cavity and release the active substance. Formulations suitable for vaginal administration also include pessaries, tampons, creams, gels, pastes, foams or spray formulations containing such carriers as are known in the art to be appropriate.

Formulations which may be used for transdermal or epicutaneous or dermal administration of the HPP composition include powders, sprays, ointments, pastes, creams, lotions (lottations), gels, solutions, patches (patches) and inhalants. The active ingredient may be mixed under sterile conditions with a pharmaceutically acceptable carrier, and with any desired preservatives, buffers, or propellants (propellants). Ointments, pastes, creams and gels may contain, in addition to the HPP composition, excipients such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or combinations thereof. Powders and sprays can contain, in addition to the HPP composition, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamides, or combinations of these substances. Sprays can additionally contain propellants such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons such as butane and propane.

The HPP/HPC or pharmaceutical composition thereof may alternatively be administered by aerosol. By making an aqueous aerosol, liposome formulation or solid particles containing HPP/HPC. Non-aqueous (e.g., fluorocarbon propellant) suspensions may also be used. Ultrasonic atomizers may also be used. Aqueous aerosols may be prepared by co-formulating an aqueous solution or suspension of the drug with conventional pharmaceutically acceptable carriers and stabilizers. The carrier and stabilizer may vary according to the requirements of the particular compound, but may generally include non-dissociating surfactants (tweens, Pluronics, or polyethylene glycols), non-toxic proteins such as serum globulins, sorbitol esters, oleates, lecithins, amino acids such as glycine, buffers, salts, sugars or sugar alcohols. Aerosols can generally be prepared from isotonic solutions.

Transdermal patches may also be used to deliver HPP compositions to the tumor site. Such formulations may be prepared by dissolving or dispersing the drug in a suitable vehicle. Pro-absorbant agents may also be used to increase the flux (flux) of the drug across the skin. The flow rate can be controlled by using a rate controlling membrane or by dispersing the drug in a polymer matrix or gel.

In certain embodiments, the transdermal therapeutic system comprises the polypeptide HPP/HPC as an active substance for treating a condition that its parent drug can treat, wherein the system is a spray or paint further comprising a solution in which the polypeptide HPP/HPC can be dissolved. Examples of such solutions include, but are not limited to, organic and inorganic solutions, such as water, ethanol, isopropanol, acetone, DMSO, DMF, and mixtures thereof.

In certain embodiments, a transdermal therapeutic system comprising the polypeptide HPP/HPC as an active agent for treating a condition treatable by its parent agent further comprises an article comprising an active agent-containing matrix layer and an impermeable backing layer. In certain embodiments, such an article is a patch or bandage. In certain embodiments, such articles comprise a permeable substrate facing the skin for the active agent storage compartment, wherein by controlling the rate of release, the system allows the active agent or metabolite thereof to continue to achieve optimal therapeutic blood levels to increase therapeutic efficacy and reduce side effects of the active agent or metabolite thereof.

Ophthalmic formulations, ocular ointments, powders, solutions, and the like are also included within the scope of the present invention.

Formulations useful for injectable administration comprise HPP/HPC and one or more pharmaceutically acceptable sterile isotonic aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, or sterile powders which may be reconstituted with sterile injectable solutions or dispersions just prior to use, which may contain antioxidants, buffers, bacteriostats, solutes which render the formulation isotonic with the blood of the intended subject, or suspending or thickening agents.

Examples of aqueous and non-aqueous carriers that can be used in formulations suitable for injectable administration include water, ethanol, polyols (e.g., glycerol, propylene glycol, polyethylene glycol, and the like), and suitable combinations thereof, vegetable oils, such as olive oil, and injectable organic esters, such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of coating materials such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.

Formulations for injectable administration may also contain adjuvants such as preserving, wetting, emulsifying, and dispersing agents. Prevention of the action of microorganisms can be ensured by the addition of various antibacterial and antifungal substances, for example, paraben, chlorobutanol, phenol sorbic acid, and the like. Isotonic substances, such as sugars, sodium chloride, and the like, may also preferably be included in the compositions. In addition, prolonged absorption of the injectable pharmaceutical form may be brought about by the addition of agents delaying absorption, such as aluminum monostearate and gelatin.

Injectable depot formulations (depot forms) can be made by forming microencapsulated matrices of HPPs or formed in biodegradable polymers such as polylactic-polyglycolide. Control of the rate of drug release may depend on the ratio of HPP/HPC to polymer, as well as the nature of the particular polymer used. Examples of other biodegradable polymers include poly (orthoesters) and poly (anhydrides). Depot injectable formulations can also be prepared by encapsulating HPP/HPC in liposomes or microemulsions compatible with body tissues.

In certain embodiments, a therapeutically effective dose of HPP/HPC of the polypeptide, or a pharmaceutical composition thereof, may be delivered to the site of the disease or tumor. It is well known in the pharmacological arts that precise amounts of a pharmaceutically effective dose of HPP, which depend, for example, on the activity, specific properties, pharmacokinetic properties, and bioavailability of the particular HPP/HPC, the physiological symptoms of the subject (including race, age, sex, body weight, diet, disease type and stage, overall physical symptoms, responsiveness to the particular dose and type of treatment), the nature of the pharmaceutically acceptable carrier in the formulation, the route and frequency of administration, the severity or predisposition of the disease to the pathogenic microorganism of interest, and the like, may provide the most effective result in terms of therapeutic efficacy for a particular patient, to name a few. However, the above guidelines may serve as a basis for fine-tuning treatment, e.g., determining the optimal dosage to administer, which does not exceed routine experimentation consisting of monitoring the subject and adjusting the dosage. For reference, Remington: the Science and Practice of Pharmacy (Gennaro ed.20.sup.th edition, Williams & Wilkins PA, USA) (2000).

Advantages of

Polypeptides and related compounds are generally relatively hydrophilic and thus difficult to penetrate the skin membrane barrier. When polypeptides are administered orally, polypeptides and polypeptide-related compounds are hydrolyzed by proteases in the gastrointestinal tract within minutes. In the case of injection, administration of polypeptide drugs causes pain, and in many cases, frequent hospitalization is required to treat chronic symptoms, which is costly.

In certain embodiments, because the polypeptide HPP or HPC can penetrate one or more biological barriers, the HPP or HPC can reach the site of symptom development by topical administration (e.g., topical or transdermal) without systemic administration (e.g., oral or injection). Local administration and penetration of HPP or HPC such that local concentrations of therapeutic substance (agent) or drug achievable by HPP or HPC are at the same level as when the parent drug or drug is administered systemically, but the amount or dose of HPP or HPC is much lower than the amount of parent drug or drug administered systemically; furthermore, systemic administration may not achieve higher local concentrations or require very high doses of therapeutic substances, if possible. The high local concentration of HPP or HPC or its sheared parent drug renders it more effective in treating the condition or more rapid than systemic administration of the parent drug and renders new conditions treatable or not previously treatable or observable treatable. Local administration of HPP or HPC may reduce the potential suffering of a biological subject from systemic administration, such as side reactions, gastrointestinal reactions or renal reactions associated with systemic exposure to therapeutic substances. In addition, topical administration allows HPP or HPC to pass through various biological barriers and through, for example, the general circulation, to the system, thus eliminating the need for systemic administration (e.g., injection) and avoiding the pain associated with injection administration.

In certain embodiments, the HPP or HPC or pharmaceutical composition of the present invention may be administered systemically (e.g., orally or by injection). HPPs or HPCs or the active therapeutic substances (e.g. drugs or metabolites) of HPPs or HPCs can enter the general circulation and reach the site of action of a condition at a faster rate than the parent drug. Furthermore, HPP or HPC can pass through biological barriers (e.g. the blood brain barrier) which are not penetrated if the parent drug alone is administered, and thus HPP or HPC can provide a new treatment for symptoms that were previously untreatable or not observed.

For example, the polypeptide HPP/HPC may exhibit a high penetration rate into a biological barrier (e.g., at least about 10-fold, about 50-fold, about 100-fold, about 200-fold, about 300-fold, about 1000-fold higher than when the polypeptide or polypeptide-related compound is administered alone). No or hardly any side effects (e.g. nausea, hair loss, and increased susceptibility to infection) were observed in subjects using the polypeptide HPP or HPC, whereas side effects were observed in subjects using the polypeptide at the same dose.

V. examples

V.EXAMPLES

The following examples are intended to better illustrate the claimed invention and should not be construed as limiting the invention in any way. All of the specific compositions, materials and methods described below, in whole or in part, are intended to be included within the scope of the present invention. These specific compositions, materials, and methods are not intended to limit the invention, but merely to illustrate specific examples within the scope of the invention. Those skilled in the art may develop equivalent compositions, materials, and methods without the exercise of inventive faculty, and without departing from the scope of the invention. It should be understood that many variations can be made using the methods described herein while still being encompassed within the scope of the invention. Such variations are considered by the inventors to be included within the scope of the invention.

Example 1 Process for making prodrugs from parent drugs

Preparation of HPP from parent drug containing at least one carboxyl group

In certain embodiments, a parent drug having the structure of the following structural formula F-C:

structural formula F-C

HPP which can be converted to a structure having the structure of formula L-1:

structural formula L-1

Including stereoisomers and pharmaceutically acceptable salts thereof, wherein:

F，L_1C，L_1N，L_2C，L_2N，L_4Cand L_4NAs defined above;

T_Cis a transport unit for the polypeptide HPP/HPC, e.g. T_CCan be selected from the group consisting of formula Na, formula Nb, formula Nc, formula Nd, formula Ne, formula Nf, formula Ng, formula Nh, formula Ni, formula Nj, formula Nk, formula Nl, formula Nm, formula Nn, formula No, formula Np, formula Nq, and formula Nr;

T_Nselected from the group consisting of zero (no group), H, substituted and unsubstituted alkyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocycloalkenyl, substituted and unsubstituted alkoxy, substituted and unsubstituted alkenyl, substituted and unsubstituted alkynyl, substituted and unsubstituted aryl, and substituted and unsubstituted heteroaryl.

In certain embodiments of the invention, the HPP/HPC of formula L-1 may be prepared by reacting a parent drug or derivative of a parent drug (e.g., an acid halide of the parent drug, a mixed anhydride, etc.):

structure of the product

Formula D

With a compound of structural formula E (scheme 1):

T_C-L_2C-H

structural formula E

Prepared by organic synthesis. Wherein Wc is selected from the group consisting of OH, halogen, hydrocarbonyl, and substituted hydrocarbonyloxy; F. l is_1C、L_1N、L_2C、L_2N、L_4C、L_4N、T_CAnd T_NAs defined above.

Scheme 1 preparation of HPP/HPC from parent drug (I)

In certain embodiments, HPP/HPC of formula L-1 is prepared by scheme 1 below, wherein L_4CIs C ═ O.

In certain embodiments, the parent drug having the following structural formula F:

structural formula F-N

With a compound having the following structural formula W:

structural formula W

To give HPP/HPC of formula L:

structural formula L-1

Including stereoisomers and pharmaceutically acceptable salts thereof, wherein:

F，L_1C，L_1N，L_2C，L_2N，L_4Cand L_4NAs defined above;

T_Nis a transport unit for the polypeptide HPP/HPC, e.g. T_NCan be selected from the group consisting of formula Na, formula Nb, formula Nc, formula Nd, formula Ne, formula Nf, formula Ng, formula Nh, formula Ni, formula Nj, formula Nk, formula Nl, formula Nm, formula Nn, formula No, formula Np, formula Nq, and formula Nr;

T_Cselected from the group consisting of zero (no group), H, substituted and unsubstituted alkyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocycloalkyl, substituted and unsubstituted alkoxy, substituted and unsubstituted alkenyl, substituted and unsubstituted alkynyl, substituted and unsubstituted aryl, and substituted and unsubstituted heteroaryl;

W_Ncan be selected from OH, halogen, hydrocarbyloxycarbonyl and substituted aryloxycarbonyloxy. (scheme 2)

Scheme 2 preparation from parent drug (II)

HPP/HPC

In certain embodiments, the HPP/HPC of formula L-2 may be prepared by organic synthesis, wherein unwanted reactive sites such as-COOH, -NH, may be attached to the functional unit prior to attachment of the delivery unit thereto₂-OH, or-SH. In certain embodiments, the obtained protected HPP/HPC may be further partially or fully deprotected to obtain a protected HPP/HPC or an unprotected HPP/HPC, respectively. Acetyl-valine-proline-aspartic acid (ethyl ester) -prolinePreparation of acid-arginine (diacetyl) -diethylaminoethyl ester hydrochloride

Preparation of arginine (diacetyl) -diethylaminoethyl ester: 30.8g Z-arginine was dissolved in 500ml acetone. 200ml of 20% sodium hydroxide solution was added to the reaction mixture solution. 40g of acetic anhydride was added dropwise to the reaction mixture. The mixture was stirred at room temperature for 2 h. The solvent was evaporated to dryness. The remaining mixture was extracted with 500ml of ethyl acetate. The ethyl acetate solution was washed with water (3 × 100 ml). The ethyl acetate layer was then dried over anhydrous sodium sulfate, the ethyl acetate solution was evaporated to dryness, and the residue (Z-Arg (diac) -OH, 30g) was dissolved in 300ml of acetonitrile. The mixture was cooled to 0 ℃ with an ice bath. 12g N, N-diethylaminoethanol, 2g 4-dimethylaminopyridine and 22g 1, 3-dicyclohexylcarbodiimide were added to the reaction mixture. The reaction mixture was stirred for 1h at 0 ℃ and then overnight at room temperature. The solid was removed by filtration and the solution was evaporated to dryness. The residue was extracted with ethyl acetate (2 × 250 ml). The ethyl acetate solution was washed with 5% sodium bicarbonate solution (1x500ml) and water (3x100 ml). The ethyl acetate solution was dried over anhydrous sodium sulfate, and the solution was evaporated to dryness. The residue (Z-Arg (diac) -OCH₂CH₂N(CH₂CH₃)₂28g) was dissolved in 300ml of methanol, and 2g of 10% Pd/C was added to the solution. The mixture was stirred at room temperature under hydrogen for 10 h. Filtering to remove Pd/C, evaporating the solution to dryness to obtain 22g H-Arg (diac) -OCH₂CH₂N(CH₂CH₃)₂。

Preparation of Boc-Asp (OEt) -Pro-OSu: 15g L-proline was dissolved in 300ml of 10% sodium bicarbonate solution, and 150ml of acetone and 36g of Boc-Asp (OEt) -OSu were added to the reaction mixture. The mixture was stirred at room temperature for 5 h. The mixture was washed with diethyl ether (1X300ml) and the aqueous layer was charged with 500ml ethyl acetate. The pH of the mixture was adjusted to 2.4-2.5 with ice-cooled 3N HCl. The ethyl acetate layer was collected and washed with water (3x300 ml). The organic solution was dried over anhydrous sodium sulfate, and then the solution was evaporated to dryness. The residue (Boc-Asp (OEt) -Pro-OH)25g and 11g N-hydroxysuccinimide were dissolved in 300ml of dichloromethane. The mixture was cooled to 0 ℃ and 16g of 1, 3-dicyclohexylcarbodiimide were added to the reaction mixture. The mixture was stirred at 0 ℃ for 1h and then filtered to remove the solids. The dichloromethane solution was washed with 5% sodium bicarbonate solution (1x200ml) and water (3x200 ml). The organic solution was dried over anhydrous sodium sulfate, and the solution was evaporated to dryness to give 28g of Boc-Asp (OEt) -Pro-OSu.

H-Asp(OEt)-Pro-Arg(diAc)-OCH₂CH₂N(CH₂CH₃)₂Preparation of TFA: 22gof H-Arg (diac) -OCH₂CH₂N(CH₂CH₃)₂Dissolved in 300ml of 5% NaHCO₃And (3) solution. 24g Boc-Asp (OEt) -Pro-OSu dissolved in 150ml acetone was added to the reaction mixture. The mixture was stirred at room temperature for 5 h. To the mixture was added 500ml ethyl acetate and the ethyl acetate solution was washed with water (3 × 100 ml). The organic solution was dried over anhydrous sodium sulfate, and the solution was evaporated to dryness. The residue was dissolved in 250ml of dichloromethane, and 200ml of trifluoroacetic acid was added thereto and stirred for 30min. The mixture is evaporated to dryness to obtain 32gH-Asp (OEt) -Pro-Arg (diac) -OCH₂CH₂N(CH₂CH₃)₂.2TFA。

Preparation of Ac-Val-Pro-OSu: 15g of L-proline are dissolved in 300ml of 10% aqueous sodium bicarbonate solution. 150ml of acetone and 26g of Ac-Val-OSu were added to the reaction mixture. The mixture was stirred at room temperature for 5 h. The mixture was washed with diethyl ether (1X300ml) and 500ml ethyl acetate was added to the aqueous layer. The pH of the mixture was adjusted to 2.4-2.5 with ice-cooled 3N HCl. The ethyl acetate layer was collected and washed with water (3x300 ml). The organic solution was dried over anhydrous sodium sulfate, and then the solution was evaporated to dryness. 20g of the residue (Ac-Val-Pro-OH) and 11g N-hydroxysuccinimide were dissolved in 300ml of methylene chloride. The mixture was cooled to 0 ℃ and 16g of 1, 3-dicyclohexylcarbodiimide were added to the reaction mixture. The mixture was stirred at 0 ℃ for 1h and then filtered to remove the solids. The dichloromethane solution was washed with 5% sodium bicarbonate solution (1x200ml) and water (3x200 ml). The organic solution was dried over anhydrous sodium sulfate, and the solution was evaporated to dryness to obtain 20g of Boc-Ac-Val-Pro-OSu.

Ac-Val-Pro-Asp(OEt)-Pro-Arg(diAc)-OCH₂CH₂N(CH₂CH₃)₂Preparation of HCl: 31g H-Asp (OEt) -Pro-Arg (diac) -OCH₂CH₂N(CH₂CH₃)₂2TFA dissolved in 300ml 10% NaHCO₃And (3) solution. 150ml of acetone and 15g of Ac-Val-Pro-OSu were added to the reaction mixture. The mixture was stirred at room temperature for 5 h. To the mixture was added 500ml of ethyl acetate, and the organic layer solution was washed with distilled water (3X100 ml). After drying the ethyl acetate over anhydrous sodium sulfate, the anhydrous sulfuric acid solid was removed by filtration. Dioxane (200ml) dissolved with 15g of HCl gas was added to the solution and the solid was collected and washed with diethyl ether (3X50 ml). After drying, 20g of the objective product (hygroscopic solid) was obtained. Elemental analysis: c₃₉H₆₆ClN₉O₁₁(ii) a Molecular weight: 872.45. theoretical value% C: 53.69; h: 7.62; cl: 4.06; n: 14.45 of; o: 20.17; found% C: 53.61, respectively; h: 7.67; cl: 4.10; n: 14.40, O: 20.22. MS: m/e: 836.4; m/e + 1: 836.4.

preparation of acetyl-tyrosine (acetyl) -glycine-phenylalanine-methionine diethylaminoethyl ester hydrochloride.

Methionine diethylaminoethyl ester trifluoroacetate (H-Met-OCH)₂CH₂N(CH₂CH₃)₂TFA). 25g of tert-butoxycarbonyl-methionine were dissolved in 300ml of dichloromethane. The mixture was cooled to 0 ℃ with an ice water bath. To the reaction mixture was added 12g N, N-diethylaminoethanol, 2g 4-dimethylaminopyridine and 22g 1, 3-dicyclohexylcarbodiimide. The reaction mixture was stirred at 0 ℃ for 1 hour and then at room temperature overnight. The solids were removed by filtration and the dichloromethane solution washed with 5% sodium bicarbonate (1X500ml) and water (3X100 ml). The ethyl acetate solution was dried over anhydrous sodium sulfate. The solution was evaporated to dryness. The residue [ tert-butoxycarbonyl-methionine diethylaminoethyl ester, 30g]Dissolved in 250ml of dichloromethane. 250ml of trifluoroacetic acid was added to the mixture, and the mixture was stirred for 30 minutes. The solution was evaporated to dryness. 26g of methionine diethylaminoethyl ester trifluoroacetate were obtained.

Preparation of t-butoxycarbonyl-glycine-phenylalanine-N-succinimide. 20g L-phenylalanine was dissolved in 300ml of 10% aqueous sodium bicarbonate solution. 150ml of acetone and 28g of t-butoxycarbonyl-glycine-N-succinimide were added to the reaction mixture. The mixture was stirred at room temperature for 5 hours. The mixture was washed once with 300ml of ether. 500ml of ethyl acetate were added to the aqueous layer. The pH of the mixture was adjusted to 2.4-2.5 with ice-cooled 3N HCl. The ethyl acetate layer was collected and washed three times with 300ml of water each time. The organic solution was dried over anhydrous sodium sulfate. The solution was evaporated to dryness. 22g of the residue (tert-butoxycarbonyl-glycine-phenylalanine) and 10g N-hydroxysuccinimide were dissolved in 300ml of dichloromethane. The mixture was cooled to 0 ℃.15 g of 1, 3-dicyclohexylcarbodiimide were added to the reaction mixture. The mixture was stirred at 0 ℃ for 1 hour. The solid was removed by filtration. The dichloromethane solution was washed once with 200ml of 5% sodium bicarbonate solution and three times with 200ml of water each time. The organic solution was dried over anhydrous sodium sulfate. The solution was evaporated to dryness. 25g of t-butoxycarbonyl-glycine-phenylalanine-N-succinimide were obtained.

Preparation of glycine-phenylalanine-methionine-diethylaminoethyl ester trifluoroacetate. 25g methionine-diethylaminoethyl trifluoroacetate was dissolved in 300ml of 5% sodium hydrogencarbonate solution. To the reaction mixture was added 150ml of an acetone solution containing 22g of t-butoxycarbonyl-glycine-phenylalanine-N-hydroxysuccinimide. The mixed solution was stirred at room temperature for 5 hours. 500ml of ethyl acetate were added to the mixture. The ethyl acetate solution was washed three times with 100ml of water each time. The organic solution was dried over anhydrous sodium sulfate. The solution was evaporated to dryness. The residue was dissolved in 250ml of dichloromethane. 200ml of trifluoroacetic acid was added to the mixture and stirred for 30 minutes. The mixture was evaporated to dryness. 25g of glycine-phenylalanine-methionine-diethylaminoethyl ester trifluoroacetate were obtained.

Preparation of acetyl-tyrosine (acetyl) -glycine-N-succinimide. 11g L-Glycine was dissolved in 300ml of 10% sodium bicarbonate solution. 150ml of acetone and 36g of acetyl-tyrosine (acetyl) -N-hydroxysuccinimide are added to the reaction mixture. The mixed solution was stirred at room temperature for 5 hours. The mixed solution was washed once with 300ml of ether. 500ml of ethyl acetate were added to the aqueous layer. The pH of the mixed solution was adjusted to 2.4-2.5 with ice-cooled 3N HCl. The ethyl acetate layer was collected and washed three times with 300ml of water each time. The organic solution was dried over anhydrous sodium sulfate. The organic solution was evaporated to dryness. 28g of the residue (acetyl-tyrosine (acetyl) -glycine) and 13g N-hydroxysuccinimide were dissolved in 300ml of dichloromethane. The mixed solution was cooled to 0 ℃. 18g of 1, 3-dicyclohexylcarbodiimide were added to the reaction mixture solution. The mixture was stirred at 0 ℃ for 1 hour. The solid was removed by filtration. The dichloromethane solution was washed once with 200ml of 5% sodium bicarbonate and three times with 200ml of water each time. The organic solution was dried over anhydrous sodium sulfate. The solution was evaporated to dryness. 20g of acetyl-tyrosine (acetyl) -glycine-N-succinimide are obtained.

Synthesis of acetyl-tyrosine (acetyl) -glycine-phenylalanine-methionine diethylaminoethyl ester hydrochloride. 24g of glycine-phenylalanine-methionine diethylaminoethyl trifluoroacetate were dissolved in 300ml of a 10% sodium hydrogencarbonate solution. 150ml of acetone and 15g of acetyl-tyrosine (acetyl) -glycine-N-succinimide were added to the reaction mixture solution. The mixture was stirred at room temperature for 5 hours. 500ml of ethyl acetate was added to the mixed solution. The organic layer was washed three times with 100ml of water each time. The ethyl acetate layer was dried over sodium sulfate. The sodium sulfate was removed by filtration. Dioxane solution (200ml) containing 15g of hydrochloric acid gas was added to the ethyl acetate solution. The solid was collected and washed three times with 50ml of ether each time. After drying, 18g of the target product which is hygroscopic is obtained. Solubility in water: 200 mg/ml; elemental analysis: c₃₇H₅₃ClN₆O₉S; molecular weight: 793.37. theoretical value (%): c: 56.01, respectively; h: 6.73; cl: 4.47; n: 10.59; o: 18.15 of; s: 4.04. found (%): c: 55.96, respectively; h: 6.76; cl: 4.52; n: 10.54, O: 18.19; s: 4.03. mass spectrum: m/e: 757.4, respectively; m/e + 1: 758.4.

preparation of acetyl-valine-proline-glycine-proline-arginine (diacetyl) diethylaminoethyl ester hydrochloride Preparation of preaparation of Ac-Val-Pro-Gly-Pro-Arg (diac) -OCH₂CH₂N(CH₂CH₃)₂·HCl

Preparation of t-butoxycarbonyl-glycine-proline-N-succinimide: 15g L-proline was dissolved in 300ml of 10% sodium bicarbonate solution. 150ml of acetone and 27.2g of t-butoxycarbonyl-glycine-N-succinimide were added to the reaction mixture solution. The mixture was stirred at room temperature for 5 hours. The mixture was washed once with 300ml of ether. 500ml of ethyl acetate were added to the aqueous layer. The pH of the mixed solution was adjusted to 2.4-2.5 with ice-cooled 3N HCl. The ethyl acetate layer was collected and washed three times with 300ml of water each time. The organic solution was dried over anhydrous sodium sulfate. The solvent was evaporated to dryness. 21g of the residue (tert-butoxycarbonyl-glycine-proline) and 11g N-hydroxysuccinimide were dissolved in 300ml of dichloromethane. The mixture was cooled to 0 ℃.17 g of 1, 3-dicyclohexylcarbodiimide were added to the reaction solution. The mixture was stirred at 0 ℃ for 1 hour. The solid was removed by filtration. The dichloromethane solution was washed once with 200ml of 5% aqueous sodium bicarbonate solution and three times with 200ml of water each time. The organic solution was dried over anhydrous sodium sulfate. The solvent was evaporated to dryness. 23g of tert-butoxycarbonyl-glycine-proline-N-succinimide were obtained.

Preparation of glycine-proline-arginine (diacetyl) -diethylaminoethyl ester bistrifluoroacetate salt: 22g of arginine (diacetyl) diethylaminoethyl ester are dissolved in 300ml of 5% sodium bicarbonate solution. To the reaction mixture was added 150ml of an acetone solution containing 20g of t-butoxycarbonyl-glycine-proline-N-succinimide. The mixed solution was stirred at room temperature for 5 hours. To the mixed solution was added 500ml of ethyl acetate. The ethyl acetate solution was washed three times with 100ml of water each time. The organic solution was dried over anhydrous sodium sulfate. The solvent was evaporated to dryness. The residue was dissolved in 250ml of dichloromethane. 200ml of trifluoroacetic acid was added to the mixed solution and stirred for 30 minutes. The mixed solution was evaporated to dryness. 28g of glycine-proline-arginine (diacetyl) diethylaminoethyl ester bistrifluoroacetate are obtained.

Preparation of acetyl-valine-proline-glycine-proline-arginine (diacetyl) -diethylaminoethyl ester hydrochloride: 26g of glycine-proline-arginine (diacetyl) diethylaminoethyl ester bistrifluoroacetate are dissolved in 300ml of a 10% sodium bicarbonate solution. 150ml of acetone and 15g of acetyl-valine-proline-N-succinimide are added to the reaction mixture. Mixed solutionThe solution was stirred at room temperature for 5 hours. 500ml of ethyl acetate was added to the mixed solution. The organic layer was washed three times with 100ml of water each time. The ethyl acetate layer was then dried over anhydrous sodium sulfate. The sodium sulfate was removed by filtration. Dioxane (200ml) containing 15g of hydrochloric acid gas was added to the ethyl acetate solution. The solid was collected and washed three times with 50ml of ether each time. After drying, 18g of the target product which is hygroscopic is obtained. Elemental analysis: c₃₅H₆₀ClN₉O₉(ii) a Molecular weight: 786.36. theoretical value (%) C: 53.46; h: 7.69; cl: 4.51; n: 16.03; o: 18.31 of the total weight of the alloy; found value (%) C: 53.43; h: 7.73; cl: 4.55; n: 16.01, O: 18.29. mass spectrum: m/e: 750.4; m/e + 1: 751.4.

preparation of cyclo (1, 6) -acetyl-norleucine-aspartic acid-histidine-phenylalanine-arginine (diacetyl) -tryptophan-lysine diethylaminoethyl ester hydrochloride

(Cyclo(1，6)-Ac-Nle-Asp-His-Phe-Arg(diAc)-Trp-Lys-OCH₂CH₂N(CH₂CH₃)₂·HCl)

Preparation of acetyl-norleucine-aspartic acid (9-fluorenylmethanol ester): 43g of aspartic acid (9-fluorenylmethanol) trifluoroacetate (H-Asp (OFm) -OH.TFA) and 27g of acetyl-norleucine-N-succinimide were suspended in 300ml of acetone. 300ml of 5% sodium bicarbonate solution was added to the reaction mixture. The mixed solution was stirred at room temperature overnight. The mixed solution was washed once with 300ml of ether. 500ml of ethyl acetate were added to the aqueous layer. The pH of the mixed solution was adjusted to 2.4-2.5 with ice-cooled 3N HCl. The ethyl acetate layer was collected and washed three times with 300ml of water each time. The organic solution was dried over anhydrous sodium sulfate. The solution was evaporated to dryness. 42g of acetyl-norleucine-aspartic acid (9-fluorenylmethanol ester) [ Ac-Nle-Asp (OFm) -OH ] were obtained.

Synthesis of 9-fluorenylmethoxycarbonyl-tryptophan-lysine (4-Pyoc). According to the reference (h.kunz and s.birnbach, Tetrahedron lett.,25，3567，1984；H.Kunz and R.Barthels，Angew.Chem.，Int.Ed.Engl.，22783, 1983) preparation of lysine (4-Pyoc) [ H-Lys (4-Pyoc) -OH]. 33g lysine (4-Pyoc)Suspended in 300ml of 5% sodium bicarbonate solution. 300ml of acetone and 52g of fluorenylmethoxycarbonyl-tryptophan-N-succinimide were added to the reaction mixture solution. The mixture was stirred at room temperature overnight. The mixed solution was washed once with 500ml of ether. 500ml of ethyl acetate was added to the mixture, and the pH of the mixture was adjusted to 2.2 to 2.3 with ice-cooled 3N hydrochloric acid. The ethyl acetate layer was collected and washed with water. The organic solution was dried over anhydrous sodium sulfate. The organic solution was evaporated to dryness. 55g fluorenylmethoxycarbonyl-tryptophan-lysine (4-Pyoc) [ Fmoc-Trp-Lys (4-Pyoc) -OH was obtained]。

Preparation of cyclo (1, 6) -acetyl-norleucine-aspartic acid-histidine-phenylalanine-arginine (diacetyl) -tryptophan-lysine: 100g of Wang-resin was suspended in 700ml of dimethylformamide. 50g fluorenylmethoxycarbonyl-tryptophan-lysine (4-Pyoc), 13g 1-hydroxybenzotriazole, 2g 4-dimethylaminopyridine and 12g N, N-diisopropylcarbodiimide were added. The mixed solution was stirred at room temperature overnight. The resin was collected by filtration and washed three times with dimethylformamide, 400ml each time, three times with methanol, 400ml each time, three times with dichloromethane, 400ml each time. 700ml of 20% piperidine was added to the resin. The mixture was stirred for 30 minutes. The resin was collected by filtration and washed three times with dimethylformamide, 400ml each time, three times with methanol, 400ml each time and three times with dichloromethane, 400ml each time. To the resin was added 700ml of dimethylformamide, 48g of fluorenylmethoxycarbonyl-arginine (diacetyl), 13g of 1-hydroxybenzotriazole, 35ml of triethylamine and 38g O- (benzotriazol-1-yl) -N, N, N ', N' -tetramethylurea. The mixed solution was stirred at room temperature for 2 h. The resin was collected by filtration and washed three times with dimethylformamide, 400ml each time, three times with methanol, 400ml each time, three times with dichloromethane, 400ml each time. 700ml of 20% piperidine were added to the resin. The mixture was stirred for 30 minutes. The resin was collected by filtration and washed three times with dimethylformamide, 400ml each time, three times with methanol, 400ml each time and three times with dichloromethane, 400ml each time. 700ml dimethylformamide, 39g fluorenylmethoxycarbonyl-phenylalanine, 13g 1-hydroxybenzotriazole, 35ml triethylamine and 38g O- (benzotriazol-1-yl) -N, N, N ', N' -tetramethylurea were added to the resin. The mixed solution was stirred at room temperature for 2 hours. The resin was collected by filtration and washed three times with dimethylformamide, 400ml each time, three times with methanol, 400ml each time, three times with dichloromethane, 400ml each time. 700ml of 20% piperidine was added to the resin. The mixture was stirred for 30 minutes. The resin was collected by filtration and washed three times with dimethylformamide, 400ml each time, three times with methanol, 400ml each time and three times with dichloromethane, 400ml each time. 700ml dimethylformamide, 60g fluorenylmethoxycarbonyl-histidine (fluorenylmethoxycarbonyl), 13g 1-hydroxybenzotriazole, 35ml triethylamine and 38g O- (benzotriazol-1-yl) -N, N, N ', N' -tetramethylurea were added to the resin. The mixed solution was stirred at room temperature for 2 hours. The resin was collected by filtration and washed three times with dimethylformamide, 400ml each time, three times with methanol, 400ml each time, three times with dichloromethane, 400ml each time. 700ml of 20% piperidine was added to the resin. The mixture was stirred for 30 minutes. The resin was collected by filtration and washed three times with dimethylformamide, 400ml each time, three times with methanol, 400ml each time and three times with dichloromethane, 400ml each time. 700ml of dimethylformamide, 60g of acetyl-norleucine-aspartic acid (9-fluorenylmethanol ester), 13g of 1-hydroxybenzotriazole, 35ml of triethylamine and 38g O- (benzotriazol-1-yl) -N, N, N ', N' -tetramethylurea were added to the resin. The mixed solution was stirred at room temperature for 2 hours. The resin was collected by filtration and washed three times with dimethylformamide, 400ml each time, three times with methanol, 400ml each time, three times with dichloromethane, 400ml each time. The resin with the peptide was suspended in 700ml of dimethylformamide. To the reaction mixture was added 50g of methyl iodide (Mel). The mixed solution was stirred at room temperature for 1 hour and at 50 ℃ for 1 hour. The resin was collected by filtration and washed three times with 400ml of dimethylformamide and three times with 400ml of methanol and three times with 400ml of dichloromethane and 400ml of dichloromethane. 700ml of 30% piperidine was added to the resin. The mixture was stirred for 60 minutes. The resin was collected by filtration and washed three times with dimethylformamide, 400ml each time, three times with methanol, 400ml each time and three times with dichloromethane, 400ml each time. 700ml dimethylformamide, 13g 1-hydroxybenzotriazole, 35ml triethylamine and 38g O- (benzotriazol-1-yl) -N, N, N ', N' -tetramethylurea were added to the resin. The mixed solution was stirred at room temperature for 10 hours. The resin was collected by filtration and washed three times with dimethylformamide, 400ml each time, three times with methanol, 400ml each time, three times with dichloromethane, 400ml each time. 500ml of trifluoroacetic acid was added to the resin, and the mixed solution was stirred at room temperature for 1 hour. The resin was removed by filtration and the organic solution was evaporated to dryness. The residue was washed three times with 100ml of ether each time.

Preparation of cyclo (1, 6) -acetyl-norleucine-aspartic acid-histidine-phenylalanine-arginine (diacetyl) -tryptophan-lysine-diethylaminoethyl ester hydrochloride. 10g of cyclo (1, 6) -acetyl-norleucine-aspartic acid-histidine-phenylalanine-arginine (diacetyl) -tryptophan-lysine were dissolved in 300ml of dimethylformamide. The mixed solution was cooled to 0 ℃ with an ice water bath. 12g N, N-dimethylaminoethanol, 2g 4-dimethylaminopyridine and 22g 1, 3-dicyclohexylcarbodiimide were added to the reaction mixture solution. The reaction solution was stirred at 0 ℃ for 1 hour and at room temperature overnight. The solids were removed by filtration and the dichloromethane solution was washed once with 500ml of 5% sodium bicarbonate solution and three times with 100ml of water. The ethyl acetate solution was dried over anhydrous sodium sulfate. Dioxane (20ml) containing 2g of hydrochloric acid gas was added to the ethyl acetate solution. The solid was collected and washed three times with 100ml of ether each time. 8g of cyclo (1, 6) -acetyl-norleucine-aspartic acid-histidine-phenylalanine-arginine (diacetyl) -tryptophan-lysine-diethylaminoethyl ester hydrochloride are obtained.

Solid phase synthesis method of HPP/HPC of polypeptide

HPP/HPC of polypeptides was synthesized using chlorinated triphenyl-based resins (2-chloro-trityl Chloride resin, 4-methyl trityl Chloride resin et al.) and 2- (4-nitrobenzenesulfonyl) ethyl as protecting groups for carboxyl groups (scheme 3), where:

is a resin;

X¹selected from H, Cl, methyl and AA collection of oxy groups;

R¹，R²，---Rⁿthe side chains at positions 1, 2, -N of the amino acids in the polypeptide HPP/HPC are counted from N-to C-terminus, respectively.

The coupling agent is selected from the group consisting of HBTU/DIPEA/HOBt, TBTU/DIPEA/HOBt, BOP/DIPEA/HOBt, HATU/DIPEA/HOBt, DIC/HOB, and mixtures thereof; and is

R_TAs previously described, selected from the group consisting of substituted and unsubstituted hydrocarbyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocycloalkyl, substituted and unsubstituted hydrocarbyloxy, substituted and unsubstituted alkenyl, substituted and unsubstituted alkynyl, substituted and unsubstituted aryl, and substituted and unsubstituted heteroaryl, and the transport unit is as previously described selected from the group consisting of formula Na, formula Nb, formula Nc, formula Nd, formula Ne, formula Nf, formula Ng, formula Nh, formula Ni, formula Nj, formula Nk, formula Nl, formula Nm, formula Nn, formula No, formula Np, formula Nq, and formula Nr.

Scheme 3: solid phase synthesis using triphenylchloro resin and 2- (4-nitrobenzenesulfonyl) ethyl as protecting group for carboxyl

Unless otherwise noted, "DCM" herein refers to dichloromethane.

In certain embodiments, the polypeptide HPP/HPC is prepared using triphenylchloro resin and 9-fluorenylmethanol as protecting groups for carboxyl groups (scheme 4), whereinX¹，R¹，R²，---Rⁿ，R_TAnd the coupling reagent is as previously described.

Polypeptide HPP/HPC

Scheme 4: synthesis of polypeptide HPP/HPC with triphenylchloro resin and 9-fluorenylmethanol as carboxyl protecting groups

In certain embodiments, the polypeptide HPP/HPC is prepared with a carbonate resin and 9-fluorenylmethanol as protecting groups for the carboxyl groups (scheme 5), whereinX¹，R¹，R²，---Rⁿ，R_TAnd the coupling reagent is as previously described.

Polypeptide HPP/HPC

Scheme 5: synthesis of polypeptide HPP/HPC Using carbonate resin and 9-Fluorenylmethanol as protecting group of carboxyl group

In certain embodiments, the carbonate resin and 2- (4-nitro) HPP/HPC polypeptidePhenylsulfonyl) ethyl as a protecting group for carboxyl groups (scheme 6), whereinX¹，R¹，R²，---Rⁿ，R_TAnd the coupling reagent is as previously described.

Polypeptide HPP/HPC

Scheme 6: synthesis of polypeptide HPP/HPC Using carbonate resin and 2- (4-Nitrophenylsulfonyl) ethyl as protecting group for carboxyl group

Example 2 HPPs from polypeptides and polypeptide-related compounds penetrate human skin faster than their parent drugs in vitro

The speed of HPPS and their parent drugs across human skin was determined in modified Franz cells. The Franz cell has two cells, an upper sample cell and a lower receiving cell. The cortex (thickness: 360-.

Test compounds (2ml, 20% in 0.2M phosphate buffer, pH7.4) were added to the upper of the Franz cell. The receiving cell was filled with 10ml of 2% bovine serum albumin in normal saline and stirred at 600 rpm. The amount of test compound that passed through the skin was measured by High Performance Liquid Chromatography (HPLC). The results are shown in FIGS. 1, 2 and 3. Slope calculations for figures 1, 2 and 3 and tables 1, 2 and 3, respectively, summarize the apparent flux of the test compounds.

Since the lowest measurable apparent flux in this method is 1. mu.g/cm²H, apparent flow rate measured by mother drug is equal to or lessAt 1. mu.g/cm²The/h is considered to be unable to pass through the skin tissue. HPPs of these parent drugs (e.g., enterostatin (e.g., valine-proline-aspartic acid-proline-arginine (VPDPR), valine-proline-glycine-proline-arginine (VPGPR), and alanine-proline-glycine-proline-Arginine (APGPR)), melanocortin II (cyclo (1, 6) -acetyl-norleucine-aspartic acid-histidine-phenylalanine-arginine-tryptophan-lysine), opioid peptides (e.g., methionine-enkephalin (tyrosine-glycine-phenylalanine-methionine), leucine-enkephalin (tyrosine-glycine-phenylalanine-leucine), tyrosine-D-alanine-glycine-N-methyl-phenylalanine-oxidized methionine alcohol, and tyrosine-D-alanine-glycine-phenylalanine-leucine)) was 1. mu.g/cm²H, so they are not detectable through skin tissue. Instead, their HPPs measured apparent flow values. Thus HPPs of polypeptides and polypeptide-related compounds cross skin tissue faster (600-fold faster) than their parent drugs.

TABLE 1 speed of HPPs and their parent drugs (I) in vitro through skin tissue

Example 3 conversion of HPP/HPC to its corresponding parent drug

HPP/HPC of polypeptides in human plasma are rapidly converted to polypeptides or polypeptide-related compounds with high conversion rates.

The polypeptide HPP/HPC (20mg) was incubated in whole blood (1ml) for 30min at 37 ℃ and then analyzed by HPLC. The results show that the HPP/HPC of most polypeptides are converted to the corresponding polypeptide or polypeptide-related compound (table 2).

TABLE 2 analysis of decomposition products of HPPs in plasma

A) Decomposition of acetyl-tyrosine (acetyl) -glycine-phenylalanine-methionine-diethylaminoethyl ester hydrochloride

Decomposition products	Measurement of
		Polypeptide HPP/HPC	3％
Acetyl-tyrosine-glycine-phenylalanine-methionine-diethylaminoethyl ester hydrochloride	2％
		Acetyl-tyrosine-glycine-phenylalanine-methionine	8％
Mother medicine	60％
		Other by-products (amino acids, dipeptides, tripeptides, tetrapeptides)	27％

B) Decomposition of N, N-dimethylaminobutylcarbonyl-tyrosine (acetyl) -glycine-phenylalanine-methionine N-butyl hydrochloride

(CH₃)₂NCH₂CH₂CH₂CO-Tyr(Ac)-Gly-Gly-Phe-Met-OCH₂CH₂CH₂CH₃，

C) Decomposition of cyclo (1, 6) -acetyl-norleucine-aspartic acid-histidine-phenylalanine-arginine (diacetyl) -tryptophan-lysine-diethylaminoethyl ester hydrochloride

EXAMPLE 4 treatment of obesity with polypeptide HPP/HPC

Enterostatin [ valine-proline-aspartic acid-arginine (VPDPR), valine-proline-glycine-proline-arginine (VPGPR), and alanine-proline-glycine-proline-Arginine (APGPR)]Is a pentapeptide derived from NH by a lipase proenzyme (procolipase)₂Truncated trypsin coenzyme, belonging to the family of gut-brain polypeptides. They regulate fat intake and are useful in the treatment of obesity (Erlangson-Albertsson C, York D, Obes. Rev.1997 Jul; 5 (4): 360-72 and Sorhede M, Mei J, Erlangson-Albertsson C., J physiol.87: 273. 275, 1993). When Osborne-Mendel rats, which were starved overnight, were injected intraperitoneally with valine-proline-aspartic acid-proline-arginine, the amount of food intake decreased in relation to the pentapeptide dose. This feeding inhibition was observed in rats fed a high-fat diet, but not in rats fed a high-carbohydrate low-fat diet (Okada s.et alol Behav.，1991 Jun；49(6)：1185-9)。

Weight-reducing effect of valine-proline-glycine-proline-arginine (nitro) isopropyl ester hydrochloride on SD rats and DB/DB mice

Valine-proline-glycine-proline-arginine (nitro) isopropyl ester hydrochloride (administered transdermally to the back of rats at a low dose of 0.3mg/kg) can reduce food intake and body weight in SD rats and DB/DB mice. The results are shown in tables 3, 4 and 5.

In the first set of experiments, 40 female SD rats (Sprague Dawley, age 15 weeks old, body weight 320g-345g divided into four groups 0.2ml of purified water was applied to the backs of rats in group a (n ═ 10) twice daily for 30 days, rats in groups B, C and D were dosed at 10mg/kg, 1mg/kg or 0.3mg/kg of valine-proline-glycine-proline-arginine (nitro) isopropyl hydrochloride, respectively, 0.2ml of an aqueous solution of valine-proline-glycine-proline-arginine (nitro) isopropyl hydrochloride was dermally administered to the backs of rats (n ═ 10), the results showed that valine-proline-glycine-proline-arginine (nitro) isopropyl ester hydrochloride was very effective in reducing body weight in rats twice daily for 30 days (table 3).

TABLE 3 weight loss effect of Val-Pro-Gly-Pro-Arg (nitro) isopropyl hydrochloride on SD rats

In the second set of experiments, 40 immature SD rats (182-223g) were divided into 4 groups. 0.2ml of purified water was smeared on the backs of group a rats (n ═ 10) twice daily for 30 days. Rats in groups B, C and D were dosed with 10mg/kg, 1mg/kg or 0.3mg/kg of valine-proline-glycine-proline-arginine (nitro) butyl ester hydrochloride, respectively, with 0.2ml of an aqueous solution of valine-proline-glycine-proline-arginine (nitro) butyl ester hydrochloride transdermally to the backs of the rats (n ═ 10) twice daily for 30 days. The results show that valine-proline-glycine-proline-arginine (nitro) butyl ester hydrochloride is very effective in controlling overweight in young rats (table 4).

TABLE 4 weight loss effect of valine-proline-glycine-proline-arginine (nitro) butyl ester hydrochloride on SD pups

In the third group of experiments, 40 obese female DB/DB mice (SLAC/DB/DB, 16 weeks old mice, body weight 55-60g) were divided into 4 groups. 0.1ml of purified water was smeared on the backs of group a rats (n ═ 10) twice daily for 30 days. Rats in groups B, C and D were dosed with 15mg/kg, 1.5mg/kg or 0.5mg/kg valine-proline-arginine (nitro) butyl ester hydrochloride, 0.1ml of an aqueous solution of valine-proline-glycine-proline-arginine (nitro) butyl ester hydrochloride was transdermally administered to the backs of the rats (n ═ 10) twice daily for 30 days. The results showed that valine-proline-glycine-proline-arginine (nitro) butyl ester hydrochloride was very effective in reducing body weight and blood glucose levels in obese mice (table 5).

TABLE 5 weight loss Effect of Val-Pro-Gly-Pro-Arg (nitro) butyl ester hydrochloride in obese mice (SLAC/DB/DB)

Val-Pro-Gly-Pro-Arg (diacetyl) -n-butyl esterWeight loss effect of hydrochloride on SD rats __

Valine-proline-glycine-proline-arginine (diacetyl) butyl ester hydrochloride (transdermal administration, rat 0.3mg/kg) reduced food intake and body weight in SD rats and DB/DB mice. The results are shown in tables 6, 7 and 8.

In the first set of experiments, 40 female SD rats (mice aged 15 weeks, weight 315-340g) were divided into 4 groups. 0.2ml of purified water was smeared on the backs of group a rats (n-10) twice daily for 30 days. Rats in groups B, C and D were dosed with 10mg/kg, 1mg/kg or 0.3mg/kg of valine-proline-glycine-proline-arginine (diacetyl) butyl ester hydrochloride, respectively, 0.2ml of an aqueous solution of valine-proline-glycine-proline-arginine (diacetyl) butyl ester hydrochloride was transdermally administered to the backs of the rats (n ═ 10) twice daily for 30 days. The results of the experiment show that valine-proline-glycine-proline-arginine (diacetyl) butyl ester hydrochloride very effectively reduces body weight in SD rats (table 6).

TABLE 6 weight loss in SD rats by Val-Pro-Gly-Pro-Arg (diacetyl) butyl ester hydrochloride

In the second set of experiments, 40 immature SD rats (180-230g) were divided into 4 groups. 0.2ml of purified water was smeared on the backs of group a rats (n ═ 10) twice daily for 30 days. Rats in groups B, C and D were dosed with 10mg/kg, 1mg/kg or 0.3mg/kg of valine-proline-glycine-proline-arginine (diacetyl) butyl ester hydrochloride, respectively, 0.2ml of an aqueous solution of valine-proline-glycine-proline-arginine (diacetyl) butyl ester hydrochloride was transdermally administered to the backs of the rats (n ═ 10) twice daily for 30 days. The results show that valine-proline-glycine-proline-arginine (diacetyl) butyl ester hydrochloride effectively controls the overweight in immature SD rats (table 7).

TABLE 7 weight loss Effect of Val-Pro-Gly-Pro-Arg (diacetyl) butyl ester hydrochloride on SD pups

In the third group of experiments, 40 female DB/DB mice (SLAC/DB/DB, 16 weeks old mice, body weight 55-60g) were divided into 4 groups. 0.1ml of purified water was smeared on the backs of group a rats (n ═ 10) twice daily for 30 days. Rats in groups B, C and D were dosed with 15mg/kg, 1.5mg/kg or 0.5mg/kg valine-proline-glycine-proline-arginine (diacetyl) butyl ester hydrochloride, respectively, 0.1ml of an aqueous solution of valine-proline-glycine-proline-arginine (diacetyl) butyl ester hydrochloride was transdermally administered to the backs of the rats (n ═ 10) twice daily for 30 days. The results indicate that valine-proline-glycine-proline-arginine (diacetyl) butyl ester hydrochloride was effective in reducing body weight and blood glucose levels in obese mice (table 6).

TABLE 8 weight loss Effect of Val-Pro-Gly-Pro-Arg (diacetyl) butyl ester hydrochloride on DB/DB mice

Weight-losing effect of alanine-proline-glycine-proline-arginine (nitro) butyl ester hydrochloride on SD rats

Alanine-proline-glycine-proline-arginine (nitro) butyl ester hydrochloride (transdermal administration, dose 0.3mg/kg) reduced food intake and body weight in SD rats and DB/DB mice. The results are shown in tables 9, 10 and 11.

In the first set of experiments, 40 female SD rats (aged 15 weeks, body weight 320-345g) were divided into 4 groups. 0.2ml of purified water was smeared on the backs of group a rats (n ═ 10) twice daily for 30 days. Rats in groups B, C and D were dosed with 10mg/kg, 1mg/kg or 0.3mg/kg alanine-proline-arginine (nitro) butyl ester hydrochloride, respectively, with 0.2ml aqueous alanine-proline-glycine-proline-arginine (nitro) butyl ester hydrochloride solution transdermally to the backs of the rats (n ═ 10) twice daily for 30 days. The results indicate that alanine-proline-glycine-proline-arginine (nitro) butyl ester hydrochloride effectively reduced body weight in SD rats (table 9).

TABLE 9 weight loss Effect of alanine-proline-glycine-proline-arginine (nitro) butyl ester hydrochloride on SD rats

In the second set of experiments, 40 immature female SD rats (182-223g) were divided into 4 groups. 0.2ml of purified water was smeared on the backs of group a rats (n ═ 10) twice daily for 30 days. Rats in groups B, C and D were dosed with 10mg/kg, 1mg/kg or 0.3mg/kg alanine-proline-arginine (nitro) butyl ester hydrochloride, 0.2ml of an aqueous solution of alanine-proline-glycine-proline-arginine (nitro) butyl ester hydrochloride was transdermally administered to the backs of the rats (n ═ 10) twice daily for 30 days. The results show that alanine-proline-glycine-proline-arginine (nitro) butyl ester hydrochloride very effectively controls the body weight of SD pups (table 10).

TABLE 10 weight loss Effect of alanine-proline-glycine-proline-arginine (nitro) butyl ester hydrochloride on immature SD rats

In the third group of experiments, 40 female DB/DB mice (SLAC/DB/DB, 16 weeks old mice, body weight 53-61g) were divided into 4 groups. 0.1ml of purified water was smeared on the backs of group a rats (n ═ 10) twice daily for 30 days. Rats in groups B, C and D were dosed with 15mg/kg, 1.5mg/kg or 0.5mg/kg alanine-proline-arginine (nitro) butyl ester hydrochloride, 0.1ml of an aqueous solution of alanine-proline-glycine-proline-arginine (nitro) butyl ester hydrochloride was transdermally administered to the backs of the rats (n ═ 10) twice daily for 30 days. The results show that alanine-proline-glycine-proline-arginine (nitro) butyl ester hydrochloride is often effective in reducing body weight and blood glucose values in obese mice (table 11).

TABLE 11 weight loss Effect of alanine-proline-glycine-proline-arginine (nitro) butyl ester hydrochloride on obese mice (SLAC/DB/DB)

Weight-reducing effect of valine-proline-aspartic acid (ethyl ester) -proline-arginine (nitro) butyl ester hydrochloride on SD rats

Valine-proline-aspartic acid (ethyl ester) -proline-arginine (nitro) butyl ester hydrochloride (transdermal administration, dose 0.3mg/kg) reduced food intake and body weight in SD rats and DB/DB mice. The results are shown in tables 12, 13 and 14.

In the first set of experiments, 40 female SD rats (aged 15 weeks, body weight 320-350g) were divided into 4 groups. 0.2ml of purified water was smeared on the backs of group a rats (n ═ 10) twice daily for 30 days. Rats in groups B, C and D were dosed with 10mg/kg, 1mg/kg or 0.3mg/kg of valine-proline-aspartic acid (ethyl ester) -proline-arginine (nitro) butyl ester hydrochloride, respectively, 0.2ml of an aqueous solution of valine-proline-aspartic acid (ethyl ester) -proline-arginine (nitro) butyl ester hydrochloride was transdermally administered to the backs of the rats (n ═ 10) twice daily for 30 days. The results showed that valine-proline-aspartic acid (ethyl) proline-arginine (nitro) butyl ester hydrochloride very effectively reduced body weight in SD rats (table 12).

TABLE 12 weight loss in SD rats by valine-proline-aspartic acid (ethyl ester) -proline-arginine (nitro) butyl ester hydrochloride

In the second set of experiments, 40 immature female SD rats (185-220g) were divided into 4 groups. 0.2ml of purified water was smeared on the backs of group a rats (n ═ 10) twice daily for 30 days. Rats in groups B, C and D were dosed with 10mg/kg, 1mg/kg or 0.3mg/kg of valine-proline-aspartic acid (ethyl ester) -proline-arginine (nitro) butyl ester hydrochloride, respectively, 0.2ml of an aqueous solution of valine-proline-aspartic acid (ethyl ester) -proline-arginine (nitro) butyl ester hydrochloride was transdermally administered to the backs of the rats (n ═ 10) twice daily for 30 days. The results showed that valine-proline-aspartic acid (ethyl) proline-arginine (nitro) butyl ester hydrochloride very effectively controlled the body weight of SD pups (table 13).

TABLE 13 weight loss Effect of valine-proline-aspartic acid (Ethyl ester) -proline-arginine (nitro) butyl ester hydrochloride on immature SD pups

In the third group of experiments, 40 female DB/DB mice (SLAC/DB/DB, 16 weeks old mice, body weight 53-61g) were divided into 4 groups. 0.1ml of water was smeared onto the backs of group a rats (n ═ 10) twice daily for 30 days. Rats in groups B, C and D were dosed with 15mg/kg, 1.5mg/kg or 0.5mg/kg valine-proline-aspartic acid (ethyl ester) -proline-arginine (nitro) butyl ester hydrochloride, respectively, 0.1ml of an aqueous solution of valine-proline-aspartic acid (ethyl ester) -proline-arginine (nitro) butyl ester hydrochloride was transdermally administered to the backs of the rats (n ═ 10) twice daily for 30 days. The results showed that valine-proline-aspartic acid (ethyl) proline-arginine (nitro) butyl ester hydrochloride very effectively reduced body weight and blood glucose values in obese mice (table 14).

TABLE 14 weight loss Effect of valine-proline-aspartic acid (ethyl) proline-arginine (nitro) butyl ester hydrochloride on obese mice (SLAC/DB/DB)

EXAMPLE 5 HPPs of Polypeptides or polypeptide-related Compounds for the treatment of erectile dysfunction and female sexual dysfunction

Melanocyte cortin ii (melanocortin ii) is a cyclic peptide of the lactam class, i.e., cyclo (1, 6) -acetyl-norleucine-aspartic acid-histidine-phenylalanine-arginine-tryptophan-lysine. It is a new class of drug candidates from the company Palatin (AMEX: PTN) for the treatment of male and female sexual dysfunction. As the first drug of a new class of drugs, melanocyte corticoid agonists, melanocyte corticoid II is expected to be effective in the treatment of male Erectile Dysfunction (ED) and female sexual dysfunction without the cardiovascular side effects common to other ED drugs. Melanocyte corticoid II does not act directly on the vascular system but acts through the central nervous system. Therefore, its safety and effectiveness are significantly superior to current products.

HPP of melanocyte cortin passes through human skin at very fast speed (-0.3-0.5 mg/h/cm)²) Also, a method for treating erectile dysfunction or improving female sexual desire with little side effect is provided.

2mg/kg cyclo (1, 6) -acetyl-norleucine-aspartic acid-histidine-phenylalanine-arginine (diacetyl) -tryptophan-lysine diethylaminoethyl ester hydrochloride (peptide A) dissolved in 0.2ml phosphate buffer solution (0.1M) at pH 7.0 was administered to the back of male rats (group A-1, 30 rats) once daily for 5 days. The same dose of cyclo (1, 6) -acetyl-norleucine-aspartic acid-histidine-phenylalanine-arginine (nitro) -tryptophan-lysine-diethylaminoethyl ester hydrochloride (polypeptide B) was administered to the backs of male rats (group B-1, 30 rats). The control rats did not receive any drug. The results showed that group A-1 had 5 more sexual requests and 3 more matings compared to the control group. Group B-1 compared to the control group, sexual requests increased by 6 pairs and matings increased by 3 pairs (Table 14).

2mg/kg of polypeptide A and polypeptide B dissolved in 0.2ml of phosphate buffered saline (0.1M) pH 7.0 was administered to the backs of male rats (30) and female rats (30) once a day for 5 days. Rats in the control group did not receive any drug. The results show that sexual challenge was increased by 6 pairs and mating was increased by 5 pairs for both group a-2 and group B-2 compared to the control group (table 14).

TABLE 14 increasing sexual challenge and mating effects following administration of HPP to melanocyte cortin in rats

	Control group	Polypeptide A	Polypeptide A	Polypeptide B	Polypeptide B
						Male rat		X	X	X	X
Female rat			X		X
						Increased number of sexual requests	1	5	6	6	6
Increased number of matings	1	3	5	3	5

X: : HPP (2mg/kg in 0.2ml phosphate buffered saline pH 7.0 (0.1M)) was administered to the back once a day for 5 days.

EXAMPLE 6 inhibition of writhing by prodrugs of enkephalins and related compounds

Opioid peptides (such as methionine-enkephalin (tyrosine-glycine-phenylalanine-methionine), leucine-enkephalin (tyrosine-glycine-phenylalanine-leucine), tyrosine-D-alanine-glycine-N-methyl-phenylalanine-oxidized methionine, and tyrosine-D-alanine-glycine-phenylalanine-leucine) exhibit morphine-like analgesic effects. After the mice were injected with acetic acid solution, the number of writhing was counted, and the inhibition rate was calculated from the blank group. Tyrosine (acetyl) -D-alanine-glycine-phenylalanine-leucine n-hexyl ester hydrochloride (10mg/kg, B), acetyl-tyrosine (acetyl) -D-alanine-glycine-phenylalanine-leucine diethylaminoethyl ester hydrochloride (10mg/kg, C) and tyrosine (acetyl) -D-alanine-glycine-phenylalanine-methionine (oxy) hydrochloride (10mg/kg, D) were transdermally administered to the neck of the mice 30 minutes before injection of the acetic acid solution. Group A is a control group. The results are shown in Table 15.

TABLE 15 inhibition of writhing by prodrugs of enkephalins and related compounds

Group of	Dosage (mg/kg)	Number of times of body twisting	％
				A	0	35.0	-
B	10	8.6	75
				C	10	5.2	85
D	10	3.2	91

The parent peptides corresponding to the HPPs in the examples are listed in Table D below:

table d parent peptides of HPPs appearing in the examples

Claims

1. A high penetration composition of a polypeptide or polypeptide-related compound comprising:

a) a functional unit;

b) a linker;

c) a transmission unit;

wherein the functional unit is covalently linked to the delivery unit via the linker;

wherein the functional unit contains a portion of an antimicrobial agent or antimicrobial related compound;

wherein the transport unit contains a protonatable amino group; and

wherein the linker comprises a chemical bond which is cleavable after the hpc crosses a biological barrier.

2. The high penetration composition according to claim 1, wherein the chemical bond is selected from the group consisting of a covalent chemical bond, an ether bond, a thioether bond, an amide bond, an ester bond, a thioester bond, a carbonate bond, a carbamate bond, a phosphate bond and an oxime bond.

3. The high penetration composition according to claim 1, wherein the portion of the polypeptide or polypeptide-related compound is converted to the polypeptide or polypeptide-related compound after cleavage of the cleavable bond.

4. The high penetration composition according to claim 1, wherein the functional unit comprises a lipophilic derivative of a moiety of the polypeptide or polypeptide-related compound.

5. The high penetration composition according to claim 4, wherein the lipophilic derivative is selected from the group consisting of carbonates, esters, amides, carbamates, N-mannich bases, ethers, thioethers, thioesters, phosphates, oximes and imines.

6. The hpc of claim 1, wherein the polypeptide or polypeptide-related compound is selected from the group consisting of a polypeptide, a polypeptide metabolite, and a therapeutic substance that can be metabolized into a polypeptide or a polypeptide metabolite, and analogs thereof.

7. The high penetration composition according to claim 1, wherein the protonatable amino group is selected from the group consisting of substituted and unsubstituted primary amino groups, substituted and unsubstituted secondary amino groups, and substituted and unsubstituted tertiary amino groups.

8. The high penetration composition according to claim 1, wherein the protonatable amino group is selected from the group consisting of formula Na, formula Nb, formula Nc, formula Nd, formula Ne, formula Nf, formula Ng, formula Nh, formula Ni, formula Nj, formula Nk, formula Nl, formula Nm, formula Nn, formula No, formula Np, formula Nq, and formula Nr:

including stereoisomers and pharmaceutically acceptable salts thereof, wherein:

R₁₁-R₁₆each independently selected from the group consisting of: none (no radical), H, CH₂COOR₁₁Substituted and unsubstituted alkyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocycloalkyl, substituted and unsubstituted aryl, substituted and unsubstituted heteroaryl, substituted and unsubstituted alkoxy, substituted and unsubstituted alkylthio, substituted and unsubstituted alkylamino, substituted and unsubstituted perfluoroalkyl, substituted and unsubstituted haloalkyl, wherein any carbon or hydrogen atom may be further replaced by O, S, P, NR, respectively₁₁Or any other pharmaceutically acceptable group;

9. a high penetration composition having the chemical structure:

structural formula L-1

Including stereoisomers and pharmaceutically acceptable salts thereof, wherein

F comprises a polypeptide or polypeptide-related compound having a structure represented by structural formula F-1:

structural formula F-1

Each A₁-A_mEach independently selected from the group consisting of: 2-naphthylalanine, substituted and unsubstituted alkyl groups, substituted and unsubstituted cycloalkyl groups, substituted and unsubstituted heterocycloalkenyl groups, substituted and unsubstituted alkoxy groups, substituted and unsubstituted alkenyl groups, substituted and unsubstituted alkynyl groups, substituted and unsubstituted alkylthio groups, substituted and unsubstituted alkylamino groups, substituted and unsubstituted perfluoroalkyl groups, substituted and unsubstituted haloalkyl groups, substituted and unsubstituted aryl groups, substituted and unsubstituted heteroaryl groups, structural formula a and structural formula B:

structural formula A; structural formula B;

each A₁-A_mEach p of (a) is independently selected from an integer;

each A₁-A_mZ on each carbon atom of_A-1Each A₁-A_mZ of (A) to_A-2，，Z_NT，Z_CT-1And Z_CT-2Each independently selected from the group consisting of: h, CH₃，C₂H₅，C₃H₇，CF₃，C₂F₅，C₃F₇Substituted and unsubstituted alkyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocycloalkyl, substituted and unsubstituted alkoxy, substituted and unsubstituted alkenyl, substituted and unsubstituted alkynyl, substituted and unsubstituted aryl, substituted and unsubstituted alkenyl, substituted and unsubstituted alkynyl, substituted and unsubstituted aryl, substituted and unsubstituted alkynyl, substitutedAnd unsubstituted heteroaryl, substituted and unsubstituted alkylthio, substituted and unsubstituted alkylamino, substituted and unsubstituted perfluoroalkyl, and substituted and unsubstituted haloalkyl;

each A₁-A_mR on each carbon atom of_AEach A₁-A_mR on each carbon atom of_B，R_NTAnd R_CTSubstituted and unsubstituted imidazolyl, substituted and unsubstituted guanidino, substituted and unsubstituted carboxy, substituted and unsubstituted carboxamide, substituted and unsubstituted alkyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocycloalkenyl, substituted and unsubstituted hydrocarbyloxy, substituted and unsubstituted hydrocarbylthio, substituted and unsubstituted hydrocarbylamino, substituted and unsubstituted hydrocarbonyl, substituted and unsubstituted perfluoroalkyl, substituted and unsubstituted haloalkyl, substituted and unsubstituted aryl, and substituted and unsubstituted heteroaryl, each independently selected from the group consisting of;

when a certain A₁-A_mR on each carbon atom when p is an integer less than 2_AOr R_BZ, which may be identical or different, on each carbon atom_A-1May be the same or different;

the amino and carboxyl groups on the polypeptide chain may further form lactam bridges;

the thiol group may further form a disulfide bridge;

each A₁-A_mT of_B，T_cAnd T_NEach independently selected from the group consisting of: (iii) No (No group), H, substituted and unsubstituted hydrocarbyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocycloalkenyl, substituted and unsubstituted hydrocarbyloxy, substituted and unsubstituted alkenyl, substituted and unsubstituted alkynyl, substituted and unsubstituted aryl, substituted and unsubstituted heteroaryl, structural formula Na, structural formula Nb, structural formula Nc, structural formula Nd, structural formula Ne, structural formula Nf, structural formula Ng, structural formula Nh, structural formula Ni, structural formula Nj, structural formula Nk, structural formula Nl, structural formula Nm, structural formula Nn, structural formula No, structural formula Np, structural formula Nq, and structural formula Nr;

structural formula Na structural formula Nb

Formula Nc formula Nd

Structural formula Ne structural formula Nf

Structural formula Ng structural formula Nh

Structural formula Ni structural formula Nj

Structural formula Nk structural formula Nl

Formula Nm formula Nn

Structural formula No structural formula Np

Structural formula Nq structural formula Nr

Each R₁₁-R₁₆Each independently selected from the group consisting of H, CH, and₂COOR₁₁substituted and unsubstituted alkyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocycloalkyl, substituted and unsubstituted aryl, substituted and unsubstituted heteroaryl, substituted and unsubstituted alkoxy, substituted and unsubstituted alkylthio, substituted and unsubstituted alkylamino, substituted and unsubstituted perfluoroalkyl, and substituted and unsubstituted haloalkyl, wherein any carbon or hydrogen atom may be further replaced by O, S, P, NR, respectively₁₁Or any other pharmaceutically acceptable group;

each A₁-A_mL of_1B，L_1CAnd L_1NSelected from the group consisting of none (no atoms), O, S, -N (L)₃)-，-N(L₃)-CH₂-O，-N(L₃)-CH₂-N(L₅)-，-O-CH₂-O-，-O-CH(L₃)-O，-S-CH(L₃)-O-；

Each A₁-A_mUpper L_2B，L_2CAnd L_2NEach selected from the group consisting of: none (none)

Each L_1B，L_1C，L_1N，L_2B，L_2C，L_2N，L_4B，L_4CAnd L₄L of₃And L₅Each independently selected from the group consisting of: none (no radical), H, CH₂COOL₆Taking and not takingSubstituted hydrocarbyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocycloalkyl, substituted and unsubstituted aryl, substituted and unsubstituted heteroaryl, substituted and unsubstituted alkoxy, substituted and unsubstituted alkylthio, substituted and unsubstituted alkylamino, substituted and unsubstituted perfluoroalkyl, and substituted and unsubstituted haloalkyl, wherein any carbon or hydrogen atom may be further replaced by O, S, P, NL, respectively₃Or any other pharmaceutically acceptable group;

L₆selected from the group consisting of: h, OH, Cl, F, Br, I, substituted and unsubstituted hydrocarbyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocycloalkyl, substituted and unsubstituted aryl, substituted and unsubstituted heteroaryl, substituted and unsubstituted hydrocarbyloxy, substituted and unsubstituted hydrocarbylthio, substituted and unsubstituted hydrocarbylamino, substituted and unsubstituted perfluoroalkyl, and substituted and unsubstituted halohydrocarbyl, wherein any carbon or hydrogen atom may be further replaced with O, S, P, P (O) OL, respectively₆，CH＝CH，C≡C，CHL₆，CL₆L₇Aryl, heteroaryl, or cyclic group substitution;

L₇selected from the group consisting of: h, OH, Cl, F, Br, I, substituted and unsubstituted alkyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocycloalkyl, substituted and unsubstituted aryl, substituted and unsubstituted heteroaryl, substituted and unsubstituted alkoxy, substituted and unsubstituted alkylthio, substituted and unsubstituted alkylamino, substituted and unsubstituted perfluoroalkyl, substituted and unsubstituted haloalkyl, wherein any carbon or hydrogen atom may be further replaced by O, S, P, P (O) OL, respectively₆，CH＝CH，C≡C，CHL₆，CL₆L₇Aryl, heteroaryl, or cyclic group.

10. The high penetration composition of claim 9 wherein the polypeptide is selected from the group consisting of: enterostatin, melanocyte cortin II, and opioid peptide.

11. The high penetration composition of claim 9 comprising a member selected from the group consisting of the following structural formulae: structural formula 2, structural formula 3, structural formula 4, structural formula 5, structural formula 6, structural formula 7, structural formula 8, structural formula 9, structural formula 10, structural formula 11, structural formula 12, structural formula 13, structural formula 14, structural formula 15, structural formula 16, structural formula 17, structural formula 18, structural formula 19, structural formula 20, structural formula 21, structural formula 22, structural formula 23, structural formula 24, structural formula 25, structural formula 26, structural formula 27, structural formula 28, structural formula 29, structural formula 30, structural formula 31, structural formula 32, structural formula 33, structural formula 34, structural formula 35, structural formula 36, structural formula 37, structural formula 38, structural formula 39, structural formula 40, structural formula 41, structural formula 42, structural formula 43, structural formula 44, structural formula 45, structural formula 46, structural formula 47, structural formula 48, structural formula 49, structural formula 50, structural formula 51, structural formula 52, structural formula 53, structural formula 54, structural formula 55, structural formula 56, structural formula 57, structural formula 58, structural formula 59, structural formula 60, structural formula 61, structural formula 62, structural formula 63, structural formula 64, structural formula 65, structural formula 66, structural formula 67, and structural formula 68, structural formula 69, structural formula 70, structural formula 71, structural formula 72, structural formula 73, structural formula 74, structural formula 75, structural formula 76, structural formula 77, structural formula 78, structural formula 79, structural formula 80, structural formula 81, structural formula 82, structural formula 83, structural formula 84, structural formula 85, structural formula 86, structural formula 87, structural formula 88, structural formula 89, structural formula 90, structural formula 91, structural formula 92, structural formula 93, structural formula 94, structural formula 95, structural formula 96, structural formula 97, structural formula 98, structural formula 99, structural formula 100, structural formula 101, structural formula 102, structural formula 103, structural formula 104, structural formula 105, the structural formula 106, the structural formula 107, the structural formula 108, the structural formula 109, the structural formula 110, the structural formula 111, the structural formula 112, the structural formula 113, the structural formula 114, the structural formula 115, the structural formula 116, the structural formula 117, the structural formula 118, the structural formula 119, the structural formula 120, the structural formula 121, the structural formula 122, the structural formula 123, the structural formula 124, the structural formula 125, the structural formula 126, the structural formula 127, the structural formula 128, the structural formula 129, the structural formula 130, the structural formula 131, the structural formula 132, the structural formula 133, the structural formula 134, the structural formula 135, the structural formula 136, the structural formula 137, the structural formula 138, the structural formula 139, the structural formula 140, the structural formula 141, the structural formula 142, the structural formula 143, the structural formula 144, the structural formula 145, the structural formula 146, the structural, the structural formula 158, the structural formula 159, the structural formula 160, the structural formula 161, the structural formula 162, the structural formula 163, the structural formula 164, the structural formula 165, the structural formula 166, the structural formula 167, the structural formula 168, the structural formula 169, the structural formula 170, the structural formula 171, the structural formula 172, the structural formula 173, the structural formula 174, the structural formula 175, the structural formula 176, the structural formula 177, the structural formula 178, the structural formula 179, the structural formula 180, the structural formula 181, the structural formula 182, the structural formula 183, the structural formula 184, the structural formula 185, the structural formula 186, the structural formula 187, the structural formula 188, the structural formula 189, the structural formula 190, the structural formula 191, the structural formula 192, the structural formula 193, the structural formula 194, the structural formula 195, the structural formula 196, the structural formula 197, the structural formula 198, the structural, structural formula 210, structural formula 211, structural formula 212, structural formula 213, structural formula 214, structural formula 215, structural formula 216, structural formula 217, structural formula 218, structural formula 219, structural formula 220, structural formula 221, structural formula 222, structural formula 223, structural formula 224, structural formula 225, structural formula 226, structural formula 227, structural formula 228, structural formula 229, structural formula 230, structural formula 231, structural formula 232, structural formula 233, structural formula 234, structural formula 235, structural formula 236, structural formula 237, structural formula 238, structural formula 239, structural formula 240, structural formula 241, structural formula 242, structural formula 243, structural formula 244, structural formula 245, structural formula 246, structural formula 247, structural formula 248, structural formula 249, structural formula 250, structural formula 251, structural formula 252, structural formula 253, structural formula 254, structural formula 255, structural formula 256, structural formula 257, structural formula 258, structural formula 259, structural formula 260, structural formula 261, structural formula 262, structural formula 263, structural formula 264, structural formula 265, structural formula 266, structural formula 267, structural formula 268, structural formula 269, structural formula 270, structural formula 271, structural formula 272, structural formula 273, structural formula 274, structural formula 275, structural formula 276, structural formula 277, structural formula 278, structural formula 279, structural formula 280, structural formula 281, structural formula 282, structural formula 283, structural formula 284, structural formula 285, structural formula 286, structural formula 287, structural formula 288, structural formula 289, structural formula 290, structural formula 291, structural formula 292, structural formula 293, structural formula 294, structural formula 295, structural formula 296, structural formula 297, structural formula 298, structural formula 299, structural formula 300, structural formula 301, structural formula 302, structural formula 303, structural formula 304, structural formula 305, structural formula 306, structural formula 307, structural formula 308, structural formula 309, structural formula 310, structural formula 311, structural formula 312, structural formula 313, structural formula 314, structural formula 315, structural formula 316, structural formula 317, structural formula 318, structural formula 319, structural formula 320, structural formula 321, structural formula 322, structural formula 323, structural formula 324, structural formula 325, structural formula 326, structural formula 327, structural formula 328, structural formula 329, structural formula 330, structural formula 331, structural formula 332, structural formula 333, structural formula 334, structural formula 335, structural formula 336, structural formula 337, structural formula 338, structural formula 339, structural formula 340, structural formula 341, and structural formula 342,

including stereoisomers and pharmaceutically acceptable salts thereof, wherein:

r is selected from the group consisting of: h, substituted and unsubstituted alkyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocycloalkyl, substituted and unsubstituted alkoxy, substituted and unsubstituted alkylthio, substituted and unsubstituted aryl, and substituted and unsubstituted heteroaryl;

X，X₄，X₅，X₆，X₇，X₈，X₉，X₁₀，X₂₁，X₂₂，X₂₃，X₂₄，X₂₅，X₂₆and X₂₇Each independently selected from the group consisting of: c ═ O, COO, CH₂OCO，COOCH₂OCO，COCH₂OCO，CH₂-O-CH(CH₂OR₄)₂，CH₂-O-CH(CH₂OCOR₄)₂，SO₂，PO(OR)，NO，O，S，NR₅And none (no group);

R₁，R₂，R₄，R₅，R₆，R₇，R₈，R₉，R₁₀，R₂₁，R₂₂，R₂₃，R₂₄，R₂₅，R₂₆and R₂₇Each independently selected from the group consisting of: h, O, NO₂Substituted and unsubstituted alkyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocycloalkyl, substituted and unsubstituted alkoxy, substituted and unsubstituted alkylthio, substituted and unsubstituted alkylamino, substituted and unsubstituted alkenyl, substituted and unsubstituted alkynyl, substituted and unsubstituted aryl, and substituted and unsubstituted heteroaryl; substituted and unsubstituted aryl, substituted and unsubstituted heteroaryl, substituted and unsubstituted alkoxy, substituted and unsubstituted alkylthio, substituted and unsubstituted alkylamino, substituted and unsubstituted perfluoroalkyl, substituted and unsubstituted haloalkyl,

ar is selected from the group consisting of phenyl, 2' naphthyl, 4-iodophenyl, substituted and unsubstituted aryl, and substituted and unsubstituted heteroaryl; and is

HA is selected from the group consisting of: HCl, HBr, HI, nitric acid, sulfuric acid, sulfurous acid, phosphoric acid, phosphorous acid, phosphonic acid, isonicotinic acid, lactic acid, salicylic acid, citric acid, tartaric acid, pantothenic acid, acid tartaric acid, ascorbic acid, succinic acid, maleic acid, gentisic acid (gentisic acid), fumaric acid, gluconic acid, glucuronic acid, saccharic acid, formic acid, benzoic acid, glutamic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid and pamoic acid.

12. The high penetration composition of claim 9 comprising a member selected from the group consisting of the following structural formulae: structural formula 1a, structural formula 1b, structural formula 1c, structural formula 1d, structural formula 1e, structural formula 1f, structural formula 1g and structural formula 1 h;

structural formula 1a

Structural formula 1b

Structural formula 1c

Structural formula 1d

Structural formula 1e

Structural formula 1f

Structural formula 1g

Structural formula 1h

Including stereoisomers and pharmaceutically acceptable salts thereof, wherein:

X₄，X₅，X₆，X₇，X₈，X₉，R₄，R₅，R₆，R₇，R₈，R₉and HA is as defined in claim 11.

13. A pharmaceutical composition comprising a high penetration composition according to claim 9 and a pharmaceutically acceptable carrier.

14. The pharmaceutical composition of claim 13, wherein the pharmaceutically acceptable carrier is polar.

15. The pharmaceutical composition of claim 13, wherein the pharmaceutically acceptable carrier is selected from the group consisting of: alcohols, ketones, esters, water, and aqueous solutions.

16. A method of penetrating a biological barrier comprising administering the pharmaceutical composition of claim 13 to the biological barrier.

17. A method for screening HPP for a desired property against a polypeptide or polypeptide-related compound, comprising the steps of:

1) covalently linking a functional unit comprising a polypeptide or polypeptide-related compound to a delivery unit via a linker to form a test composition;

2) administering the test composition to a biological system; and

3) determining whether the test composition has the desired property or characteristic

18. The method of claim 17, wherein the desired property is selected from the group consisting of:

1) the ability of the test composition to penetrate the biological barrier;

2) the ability of the test composition to convert to a parent drug or active therapeutic substance;

3) the penetration rate of the test composition;

4) the efficiency of the test composition; and

5) the effect of the composition was tested.

19. A method of diagnosing a condition in a biological subject, comprising the steps of:

1) administering to the biological subject any of the compositions described in claim 9;

2) detecting the presence, location or amount of said composition in the biological subject; and

3) detecting a symptom in the biological subject.

20. The method of claim 19, wherein the composition is labeled.

21. A method of diagnosing a condition in a biological subject, comprising the steps of:

1) administering to the biological subject any of the compositions of claim 13;

3) detecting a symptom in the biological subject.

22. The method of claim 21, wherein the composition is labeled.

23. A method of treating a condition in a biological subject comprising administering to said biological subject a hpc according to claim 9 or a pharmaceutical composition according to claim 13.

24. The method of claim 23, wherein the condition is selected from the group consisting of pain, injury, inflammation-related condition, microorganism-related condition, neuropeptide-related condition, hormone-related condition, tumor, blood pressure abnormality, obesity, brain injury, allergy, male and female sexual dysfunction, metastatic tumor, and other conditions associated with a deglutition hormone, prenatal, postnatal, anti-AD effect, anti-diuretic effect, calcium balance, melanocytes, hormone release, platelet aggregation, central nervous system activity, and phagocytosis.

25. The method of claim 24, wherein the hormone-related condition is selected from the group consisting of menopause, bone disease, growth hormone deficiency, hyperthyroidism, hypothyroidism, metabolic market, blood pressure abnormalities, skin disorders, autoimmune disease, eye disorders, pre-eclamptic toxemia in high risk women, male and female sexual dysfunction, allergies, asthma, insomnia, depression and related conditions, cardiovascular disease, and tumors.

26. A method according to claim 25 wherein the bone disease is selected from the group consisting of osteoporosis, pelott's disease and bone metastases.

27. The method of claim 25, wherein the symptom of metabolic abnormality is selected from the group consisting of obesity, blood glucose abnormality, dyslipidemia, diabetes (type I or/and type II) and diabetes-induced syndrome including diabetic retinopathy, necrotic ulcers, and diabetic proteinuria.

28. The method of claim 25, wherein the blood pressure abnormality is selected from the group consisting of hypertension and hypotension.

29. The method of claim 25, wherein the dermatological disorder is selected from the group consisting of psoriasis and leukemia, comedones, cystic acne, abscesses or inflamed lesions, comedones, papules, pustules, nodules, epidermoid cysts, keratosis pilaris, abnormalities of cutaneous lesions of the venous vasculature, birthmarks, nevi, cutaneous prolapse, scleroderma, vitiligo and related disorders, and age spots (liver spots).

30. The method of claim 25, wherein the autoimmune disease is selected from the group consisting of discoid lupus erythematosus, Systemic Lupus Erythematosus (SLE), autoimmune liver disease, scleroderma, sjogren's syndrome, rheumatoid arthritis, polymyositis, scleroderma, hashimoto's thyroiditis, juvenile diabetes, edison's disease, vitiligo, pernicious anemia, glomerulonephritis, pulmonary fibrosis, Multiple Sclerosis (MS), and crohn's disease.

31. The method of claim 25 wherein the ocular disease is selected from the group consisting of impaired vision in warm-blooded animals caused by glaucoma, ocular hypertension, blindness following ophthalmic surgery, cystoid macular edema, and cataracts.

32. The method of claim 25, wherein the cardiovascular disorder is selected from the group consisting of myocardial infarction, unstable angina, peripheral arterial occlusive disorder, and stroke;

33. the method of claim 25, wherein the tumor is selected from the group consisting of benign tumors, breast cancer, colon cancer, oral cancer, lung or respiratory tract cancer thereof, skin cancer, it, uterine cancer, pancreatic cancer, prostate cancer, vaginal cancer, urinary organ cancer, leukemia and other cancers of the blood and lymphoid tissues.

34. The method of claim 25, wherein the microbe-associated condition is selected from the group consisting of pain, injury, and inflammation-associated conditions.

35. The method of claim 34, wherein the symptoms associated with inflammation are selected from the group consisting of prostatitis, prostacystitis, prostatic hypertrophy fibrosis, hemorrhoids, kawasaki disease, gastroenteritis, type I membranoproliferative glomerulonephritis, bart's syndrome, chronic uveitis, ankylosing spondylitis, hemophilia arthritis, inflammatory hemorrhoids, radiation proctitis, chronic ulcerative colitis, inflammatory bowel disease, pouchitis, periodontitis, arthritis, and inflammatory diseases of the liver, lung, stomach, brain, kidney, heart, ear, eye, nose, mouth, tongue, colon, pancreas, gall bladder, duodenum, gastrorectum, colorectal, intestine, vein, respiratory system, vasculature, anorectum, and anal itch.

36. The method of claim 24, wherein the neuropeptide-related condition is selected from the group consisting of senile dementia and parkinson's disease.

37. The method of claim 23, wherein the route of administration of the composition to the organism is selected from the group consisting of oral, enteral, oral, nasal, topical, rectal, vaginal, aerosol, transmucosal, epithelial, ocular, pulmonary, subcutaneous, and injectable.

38. The method of claim 23, wherein the polypeptide is selected from the group consisting of angiotensin, angiotensin II antagonist, angiotensin II AT2 receptor, antimicrobial peptide, oxytocin, hormone, antidiuretic hormone, adrenocorticotropic hormone, anti-inflammatory peptide, bradykinin antagonist, endothelin peptide antagonist, gastrin, calcitonin, melanoma-associated antigenic peptide, fibronectin peptide, fibrinogen peptide, EAE-inducing peptide, growth factor, growth hormone releasing peptide, somatostatin, hormone releasing hormone, luteinizing hormone releasing hormone, neuropeptide, melanocyte stimulating peptide, sleep inducing peptide, amyloid peptide, deglutition inducing hormone, retroverterso-deglutition inducing hormone, enterostatin, melanocortin II, opioid peptide and mimetic.

39. The method of claim 38, wherein the enterostatin is selected from the group consisting of valine-proline-aspartic acid-proline-arginine (VPDPR), valine-proline-glycine-proline-arginine (VPGPR), and alanine-proline-glycine-proline-Arginine (APGPR).

40. The method of claim 39, wherein the opioid peptide is selected from the group consisting of methionine-enkephalin (tyrosine-glycine-phenylalanine-methionine), leucine-enkephalin (tyrosine-glycine-phenylalanine-leucine), tyrosine-D-alanine-glycine-N-methyl-phenylalanine-oxidized methionine, and tyrosine-D-alanine-glycine-phenylalanine-leucine).

41. Solid phase synthesis the solid phase synthesis method of the polypeptide HPP comprises the following steps:

a) providing a chemically modified resin;

b) depending on the polypeptide HPC with protecting groups, it is necessary to protect the carboxyl groups of all natural or modified amino acids to obtain a certain COOH-protected amino acid;

c) the N-terminal COOH-protected amino acid of the polypeptide HPC is linked to a chemically modified resin to form an immobilized COOH-protected polypeptide HPC precursor with one amino acid;

d) deprotecting the immobilized COOH-protected polypeptide HPC with a deprotection reagent to obtain an immobilized COOH-unprotected polypeptide HPC precursor with one amino acid;

e) repeating steps C) and d) with the COOH-protected amino acids of the polypeptide HPC until an immobilized COOH-unprotected precursor polypeptide HPC comprising all but the C-terminal amino acids is synthesized;

f) linking the C-terminal amino acid and R by a covalent bond_TForming a modified C-terminal amino acid wherein R_TA combination selected from the following groups according to claim 8: substituted and unsubstituted alkyl, substituted and unsubstituted cycloalkyl, substituted and unsubstituted heterocycloalkyl, substituted and unsubstituted alkoxy, substituted and unsubstituted alkenyl, substituted and unsubstituted alkynyl, substituted and unsubstituted aryl, substituted and unsubstituted heteroaryl, and the transport unit is selected from the group consisting of: structural formula Na, structural formula Nb, structural formula Nc, structural formula Nd, structural formula Ne, structural formula Nf, structural formula Ng, structural formula Nh, structural formula Ni, structural formula Nj, structural formula Nk, structural formula Nl, structural formula Nm, structural formula Nn, structural formula No, structural formula Np, structural formula Nq and structural formula Nr;

g) attaching the modified C-terminal amino acid to the immobilized C-unprotected polypeptide HPC precursor synthesized in step e) to obtain an immobilized polypeptide HPC;

h) the immobilized polypeptide HPC is cleaved from the resin to yield the polypeptide HPC.

42. A synthesis method according to claim 41, wherein the resin used is selected from the group consisting of triphenylchloride resin and carbonate resin.

43. The synthetic method of claim 41, wherein protecting group is selected from the group consisting of 2- (4-nitrophenylsulfonyl) ethyl and 9-fluorenylcarbinol.

44. The synthetic method of claim 41, wherein the coupling reaction is carried out in the presence of a coupling agent selected from the group consisting of HBTU/DIPEA/HOBt, TBTU/DIPEA/HOBt, BOP/DIPEA/HOBt, HATU/DIPEA/HOBt, and DIC/HOB.

45. A transdermal therapeutic application system comprising as an active ingredient a compound or composition according to claim 9, 10, 11, 12, or 13 for treating a condition treatable by their parent compound, and wherein the spray or liniment further comprises a solvent selected from the group consisting of water, ethanol, isopropanol, acetone, DMSO, DMF, and mixtures thereof.

46. A transdermal therapeutic application system comprising as active ingredient a compound or composition according to claim 9, 10, 11, 12, or 13 for treating a condition treatable by their parent compound, and further comprising a matrix layer containing the active agent and a non-permeable protective layer.

47. The transdermal therapeutic application system of claim 46 which is a bandage or patch.

48. The transdermal therapeutic application system of claim 46, comprising an active agent reservoir comprising a permeable skin facing base; by controlling the release rate, the system can stabilize the active ingredient or the metabolite of the active ingredient at an optimal therapeutic blood concentration to improve the therapeutic effect and reduce the side effects of the active ingredient or the metabolite of the active ingredient.