多肽口服制剂的胃肠道吸收评价模型及其应用进展

Evaluation Models for Gastrointestinal Absorption of Oral Polypeptide Preparations and Their Application Progress

摘要

多肽药物在糖尿病、肿瘤及免疫疾病治疗中展现出重要潜力，但其口服递送面临胃肠道酶降解、黏液屏障阻碍及上皮渗透性低等多重挑战，导致生物利用度极低。与传统小分子药物相比，多肽具有分子量大、亲水性强、易被酶解等特点，因此其吸收评价模型需具备更高的生理相关性与机制针对性。综述当前适用于多肽口服制剂评价的各类模型，包括细胞模型、离体肠囊模型、在体模型及动物模型，重点分析各类模型在多肽药物吸收评价中的适用性，并结合典型案例阐述其在口服多肽药物研发中的具体应用，以期为多肽口服制剂的合理评价与模型选择提供科学依据。

Abstract

Peptide drugs have demonstrated significant potential in the treatment of diabetes, tumors, and immune disorders, however, their oral delivery faces multiple challenges, including gastrointestinal enzymatic degradation, mucus barrier obstruction, and low epithelial permeability, resulting in extremely low bioavailability. Compared to traditional small-molecule drugs, polypeptides exhibit such characteristics as high molecular weight, strong hydrophilicity, and susceptibility to enzymatic degradation, thus their absorption evaluation models require higher physiological relevance and mechanistic specificity. This article reviews various models currently applicable for the evaluation of oral polypeptide preparations, including cell models, ex vivointestinal sac models, in vivomodels, and animal models, with particular focus on analyzing the applicability of these models in the absorption evaluation of peptide drugs, and illustrates their specific applications in oral polypeptide development through case studies, so as to provide a scientific basis for the rational evaluation and model selection for oral polypeptide preparations.

关键词

多肽口服制剂 / 胃肠道吸收 / 体外模型 / 在体模型 / 动物模型

Key words

oral polypeptide formulation / gastrointestinal absorption / in vitromodel / in vivomodel / animal model

参考文献

[1] Sharma K, Sharma K K, Sharma A,et al. Peptide—based drug discovery: current status and recent advances[J].Drug Discov Today, 2023, 28(2): 103464.

[2] Zhu Q, Chen Z, Paul P K,et al. Oral delivery of proteins and peptides: challenges, status quo and future perspectives[J].Acta Pharm Sin B, 2021, 11(8): 2416-2448.

[3] Wright L, Barnes T J, Prestidge C A. Oral delivery of protein—based therapeutics: gastroprotective strategies, physiological barriers and in vitropermeability prediction[J].Int J Pharm, 2020, 585: 119488.

[4] 柳梦媛, 徐晨, 姚文兵, 等. 口服多肽和蛋白质药物的研究进展[J].药学进展, 2021, 45(6): 452-459.

[5] Diehl H P, Wildey A, Prasasty V D,et al. Organization of the intestinal mucosa and barriers to oral drug delivery[M]//Nanotechnology for Oral Drug Delivery. Amsterdam: Elsevier, 2020: 7-25.

[6] Volpe D A. Advances in cell—based permeability assays to screen drugs for intestinal absorption[J].Expert Opin Drug Discov, 2020, 15(5): 539-549.

[7] Steyn J D, Haasbroek—Pheiffer A, Pheiffer W,et al. Evaluation of drug permeation enhancement by using in vitroand ex vivomodels[J].Pharmaceuticals, 2025, 18(2): 195.

[8] Jochems P G M, Garssen J, van Keulen A M,et al. Evaluating human intestinal cell lines for studying dietary protein absorption[J].Nutrients, 2018, 10(3): 322.

[9] Panse N, Gerk P M. The Caco—2 model: modifications and enhancements to improve efficiency and predictive performance[J]. Int J Pharm, 2022, 624: 122004.

[10] Birch D, Diedrichsen R G, Christophersen P C,et al. Evaluation of drug permeation under fed state conditions using mucus—covered Caco—2 cell epithelium[J].Eur J Pharm Sci, 2018, 118: 144-153.

[11] Floor E, Su J, Chatterjee M,et al. Development of a Caco—2—based intestinal mucosal model to study intestinal barrier properties and bacteria—mucus interactions[J].Gut Microbes, 2025, 17(1): 2434685.

[12] 刘瑶, 曾苏. MDCK—MDR1细胞模型及其在药物透过研究中的应用进展[J].药学学报, 2008, 43(6): 559-564.

[13] Fedi A, Vitale C, Ponschin G,et al. In vitromodels replicating the human intestinal epithelium for absorption and metabolism studies: a systematic review[J].J Control Release, 2021, 335: 247-268.

[14] Itagaki M, Kamei N, Takeda—Morishita M. Evaluation of function and features of human induced pluripotent stem cell—derived small intestinal epithelial cells for analyzing peptide drug intestinal absorption profiles[J].J Pharm Sci, 2023, 112(9): 2591-2595.

[15] Naessens U, Gutjahr A, Blache G,et al. 1677—P: AdOral: a versatile platform for improved oral peptide absorption[J].Diabetes, 2024, 73(Supplement_1): 1677—P.

[16] Diedrichsen R G, Mishra N K, Fredholt F,et al. Branched penetratin and penetramax display enhanced intestinal insulin delivery potency compared to their linear counterparts[J].J Control Release, 2025, 380: 1031-1042.

[17] 李松明, 赵梦蝶, 刘伟, 等. 卵黄高磷蛋白磷酸肽在Caco—2细胞单层模型中的促钙转运作用[J].中国食品学报, 2023, 23(6): 8-17.

[18] Berger E, Nassra M, Atgié C,et al. Oleic acid uptake reveals the rescued enterocyte phenotype of colon cancer Caco—2 by HT29—MTX cells in co—culture mode[J].Int J Mol Sci, 2017, 18(7): 1573.

[19] Cheng Y, Watanabe C, Ando Y,et al. Caco—2 cell sheet partially laminated with HT29—MTX cells as a novel in vitromodel of gut epithelium drug permeability[J].Pharmaceutics, 2023, 15(9): 2338.

[20] Antunes F, Andrade F, Araújo F,et al. Establishment of a triple co—culture in vitrocell models to study intestinal absorption of peptide drugs[J].Eur J Pharm Biopharm, 2013, 83(3): 427-435.

[21] Nagasawa M, Onuki M, Imoto N,et al. Fabrication of 3D engineered intestinal tissue producing abundant mucus by air—liquid interface culture using paper—based dual—layer scaffold[J].Biofabrication, 2024, 16(3): 035029.

[22] Estermann M, Bisig C, Septiadi D,et al. Bioprinting for human respiratory and gastrointestinal in vitromodels[M]// Crook J M.3D Bioprinting. New York: Humana, 2020: 199-217.

[23] Rudolph S E, Longo B N, Tse M W,et al. Crypt—villus scaffold architecture for bioengineering functional human intestinal epithelium[J].ACS Biomater Sci Eng, 2022, 8(11): 4942-4955.

[24] Markus J, Landry T, Stevens Z,et al. Human small intestinal organotypic culture model for drug permeation, inflammation, and toxicity assays[J].In Vitro Cell Dev Biol Anim, 2021, 57(2): 160-173.

[25] Fair K L, Colquhoun J, Hannan N R F. Intestinal organoids for modelling intestinal development and disease[J]. Philos Trans R Soc Lond B Biol Sci, 2018, 373(1750): 20170217.

[26] Wang Y, Gao Y, Pan Y,et al. Emerging trends in organ—on—a—chip systems for drug screening[J].Acta Pharm Sin B, 2023, 13(6): 2483-2509.

[27] Kim R, Sung J H. Recent advances in gut— and gut—organ—axis—on—a—chip models[J].Adv Healthc Mater, 2024, 13(21): e2302777.

[28] Lee S Y, Lee Y, Choi N,et al. Development of gut—mucus chip for intestinal absorption study[J].BioChip J, 2023, 17(2): 230-243.

[29] Lee S Y, Sung J H. Gut—on—a—chip for in vitrodrug absorption models[J].BioChip J, 2025, 19(3): 545-555.

[30] Gleeson J P, Zhang S Y, Subelzu N,et al. Head—to—head comparison of Caco—2 transwell and gut—on—a—chip models for assessing oral peptide formulations[J].Mol Pharm, 2024, 21(8): 3880-3888.

[31] Deng S, Li C, Cao J,et al. Organ—on—a—chip meets artificial intelligence in drug evaluation[J].Theranostics, 2023, 13(13): 4526-4558.

[32] Masloh S, Culot M, Gosselet F,et al. Challenges and opportunities in the oral delivery of recombinant biologics[J].Pharmaceutics, 2023, 15(5): 1415.

[33] Koziolek M, Augustijns P, Berger C,et al. Challenges in permeability assessment for oral drug product development[J].Pharmaceutics, 2023, 15(10): 2397.

[34] U. S. Food and Drug Administration, Center for Drug Evaluation and Research. Waiver of in vivobioavailability and bioequivalence studies for immediate—release solid oral dosage forms based on a biopharmaceutics classification system: guidance for industry[R]. Rockville: U. S. Department of Health and Human Services,2017.

[35] Luo Z, Liu Y, Zhao B,et al. Ex vivoand in situapproaches used to study intestinal absorption[J].J Pharmacol Toxicol Methods, 2013, 68(2): 208-216.

[36] Zhang Z, Lu Y, Qi J,et al. An update on oral drug delivery viaintestinal lymphatic transport[J].Acta Pharm Sin B, 2021, 11(8): 2449-2468.

[37] Trevaskis N L, Kaminskas L M, Porter C J H. From sewer to saviour — targeting the lymphatic system to promote drug exposure and activity[J].Nat Rev Drug Discov, 2015, 14(11): 781-803.

[38] Trevaskis N L, Hu L, Caliph S M,et al. The mesenteric lymph duct cannulated rat model: application to the assessment of intestinal lymphatic drug transport[J].J Vis Exp, 2015(97): 52389.

[39] Banan B, Wei Y, Simo O,et al. Intestinal lymph collection viacannulation of the mesenteric lymphatic duct in mice[J].J Surg Res, 2021, 260: 399-408.

[40] Sjögren E, Abrahamsson B, Augustijns P,et al. In vivomethods for drug absorption — comparative physiologies, model selection, correlations with in vitromethods (IVIVC), and applications for formulation/API/excipient characterization including food effects[J].Eur J Pharm Sci, 2014, 57: 99-151.

[41] Xu Y, Shrestha N, Préat V,et al. An overview of in vitro, ex vivoand in vivomodels for studying the transport of drugs across intestinal barriers[J].Adv Drug Deliv Rev, 2021, 175: 113795.

[42] Morgan S J, Elangbam C S, Berens S,et al. Use of animal models of human disease for nonclinical safety assessment of novel pharmaceuticals[J].Toxicol Pathol, 2013, 41(3): 508-518.

[43] Cao X, Gibbs S T, Fang L,et al. Why is it challenging to predict intestinal drug absorption and oral bioavailability in human using rat model[J].Pharm Res, 2006, 23(8): 1675-1686.

[44] Stappaerts J, Brouwers J, Annaert P,et al. In situperfusion in rodents to explore intestinal drug absorption: challenges and opportunities[J].Int J Pharm, 2015, 478(2): 665-681.

[45] Haller S, Schuler F, Lazic S E,et al. Expression profiles of metabolic enzymes and drug transporters in the liver and along the intestine of beagle dogs[J].Drug Metab Dispos, 2012, 40(8): 1603-1610.

[46] Henze L J, Koehl N J, O’Shea J P,et al. The pig as a preclinical model for predicting oral bioavailability andin vivoperformance of pharmaceutical oral dosage forms: a PEARRL review[J].J Pharm Pharmacol, 2019, 71(4): 581-602.

[47] Sinko P J, Leesman G D, Amidon G L. Mass balance approaches for estimating the intestinal absorption and metabolism of peptides and analogues: theoretical development and applications[J].Pharm Res, 1993, 10(2): 271-275.

[48] Harloff—Helleberg S, Nielsen L H, Nielsen H M. Animal models for evaluation of oral delivery of biopharmaceuticals[J].J Control Release, 2017, 268: 57-71.

[49] Buckley S T, Bækdal T A, Vegge A,et al. Transcellular stomach absorption of a derivatized glucagon—like peptide—1 receptor agonist[J].Sci Transl Med, 2018, 10(467): eaar7047.

[50] Aroda V R, Blonde L, Pratley R E. A new era for oral peptides: SNAC and the development of oral semaglutide for the treatment of type 2 diabetes[J].Rev Endocr Metab Disord, 2022, 23(5): 979-994.

[51] Tuvia S, Pelled D, Marom K,et al. A novel suspension formulation enhances intestinal absorption of macromolecules viatransient and reversible transport mechanisms[J].Pharm Res, 2014, 31(8): 2010-2021.

[52] Cheng L, Venkataraman S, Zhao L,et al. P004 PN—943, an oral α4β7 integrin antagonist, inhibits MAdCAM1—mediated proliferation and cytokine release from CD4⁺ T cells independent of trafficking[J].J Crohn’s Colitis, 2020, 14(Supplement_1): S131.

[53] Johns D G, Campeau L C, Banka P,et al. Orally bioavailable macrocyclic peptide that inhibits binding of PCSK9 to the low density lipoprotein receptor[J].Circulation, 2023, 148(2): 144-158.

[54] de Leon—Tabaldo A, Strawn D, Chang L,et al. P626 JNJ—77242113, an oral peptide selectively targeting the IL—23 receptor, demonstrates pharmacodynamic activity in rat and human colon tissue explants following oral dosing[J].J Crohn’s Colitis, 2024, 18(Supplement_1): i1193.