Pentlavalli Sreekanth, Coulter Sophie M, An Yuming, Cross Emily R, Sun Han, Moore Jessica V, Sabri Akmal Bin, Greer Brett, Vora Lalitkumar, McCarthy Helen O, Laverty Garry
School of Pharmacy, Queen's University Belfast, Medical Biology Centre, 97 Lisburn Road, Belfast, Northern Ireland BT9 7BL, United Kingdom.
School of Biological Sciences, Biological Sciences Building, 19 Chlorine Gardens, Belfast, Northern Ireland BT9 5DL, United Kingdom.
J Control Release. 2025 Mar 10;379:30-44. doi: 10.1016/j.jconrel.2024.12.052. Epub 2025 Jan 8.
New multipurpose prevention technology products for use by women, focused on reducing HIV infection and preventing unwanted pregnancies, are a global health priority. Discreet long-acting formulations will empower women with greater choice around their sexual health. This paper outlines the development of a long-acting technology that enables multiple drugs to be incorporated within one injectable platform. This fixed-dose combination product is formed from a phosphorylated D-peptide (naphthalene-2-ly)-acetyl-diphenylalanine-lysine-tyrosine-glycine-OH (Napffky(p)G-OH) that enables the highly hydrophobic drugs MIV-150 (HIV antiretroviral) and etonogestrel (contraceptive) to be solubilized together within aqueous solvents. Upon subcutaneous injection, this D-peptide-drug combination self-assembles in response to phosphatase enzymes present within the skin space to form an in situ forming drug-releasing hydrogel depot. Oscillatory rheology confirmed the formation of hydrogels, which began within ∼10 s exposure to 3.98 U/mL phosphatase enzymes and continued for ∼198 mins for a Napffk(MIV-150)y(p)G-OH + Napffk(ENG)y(p)G-OH combination (8:2 ratio). Biostability against proteases, an important consideration for long-acting injectables, was demonstrated for at least 28 days in vitro. Covalent attachment of each drug to the D-peptide via an ester linkage enabled sustained release of the drug in an unmodified form via hydrolysis of the D-peptide-drug linker. This significantly reduced the initial drug burst. Low toxicity was also demonstrated in vitro via cell culture (MTS, LHS, Live/Dead®) and within in vivo studies (H&E staining). The fixed dose combination was able to deliver clinically relevant concentrations of each drug to Sprague-Dawley rats for at least 49 days, providing proof-of-concept for the use of hydrogel-forming D-peptides (Napffky(p)G-OH) as a long-acting injectable platform for the delivery of multiple hydrophobic drugs.
供女性使用的新型多用途预防技术产品，专注于降低艾滋病毒感染率和预防意外怀孕，是全球卫生工作的重点。隐蔽的长效制剂将使女性在性健康方面有更多选择。本文概述了一种长效技术的开发，该技术能使多种药物整合在一个可注射平台内。这种固定剂量组合产品由一种磷酸化D肽（萘-2-基）-乙酰基-二苯基丙氨酸-赖氨酸-酪氨酸-甘氨酸-OH（Napffky(p)G-OH）构成，它能使高度疏水的药物MIV-150（抗艾滋病毒药物）和依托孕烯（避孕药）在水性溶剂中共同溶解。皮下注射后，这种D肽-药物组合会响应皮肤间隙中存在的磷酸酶而自组装，形成原位形成药物释放的水凝胶贮库。振荡流变学证实了水凝胶的形成，对于Napffk(MIV-150)y(p)G-OH + Napffk(ENG)y(p)G-OH组合（8:2比例），在接触3.98 U/mL磷酸酶约10秒内开始形成，并持续约198分钟。对长效注射剂来说很重要的抗蛋白酶生物稳定性在体外至少维持了28天。每种药物通过酯键与D肽共价连接，通过D肽-药物连接子的水解使药物以未修饰的形式持续释放。这显著减少了药物的初始突释。通过细胞培养（MTS、LHS、Live/Dead®）和体内研究（苏木精-伊红染色）在体外也证明了低毒性。固定剂量组合能够将每种药物的临床相关浓度输送给Sprague-Dawley大鼠至少49天，为使用形成水凝胶的D肽（Napffky(p)G-OH）作为长效注射平台来输送多种疏水药物提供了概念验证。
