TW202302525A

TW202302525A - Benzamide compounds and uses thereof

Info

Publication number: TW202302525A
Application number: TW111124360A
Authority: TW
Inventors: 祝令建; 于秀招; 蔡德勤; 管忠俊; 劉崇懿; 姜軍; 黃建
Original assignee: 大陸商江蘇恆瑞醫藥股份有限公司; 大陸商上海森輝醫藥有限公司; 大陸商上海盛迪醫藥有限公司
Priority date: 2021-06-29
Filing date: 2022-06-29
Publication date: 2023-01-16
Also published as: WO2023274265A1

Abstract

The disclosure relates to benzamide compounds and uses thereof. Specifically, a compound of formula I or a salt thereof is provided, wherein R1 to R6, M1 to M6, m1 to m3, n1 to n6 are as defined herein.

Description

苯醯胺類化合物及其用途Benzamides and uses thereof

本揭露屬於醫藥領域，關於苯醯胺類化合物及其用途。The disclosure belongs to the field of medicine, and relates to benzamide compounds and their uses.

基於核酸類的藥物，例如信使RNA（mRNA）、反義寡核苷酸、小干擾RNA (siRNA)、質粒等具有廣闊的應用前景，如何安全有效地遞送到體內靶器官和靶細胞是制約該技術發展的難題。Nucleic acid-based drugs, such as messenger RNA (mRNA), antisense oligonucleotides, small interfering RNA (siRNA), and plasmids, have broad application prospects. How to safely and effectively deliver them to target organs and cells in the body is a major constraint. Problems of technological development.

目前核酸藥物遞送系統可分為病毒載體系統和非病毒系統2大類，脂質體介導的核酸藥物遞送是屬非病毒遞送系統的主要方法。At present, nucleic acid drug delivery systems can be divided into two categories: viral vector systems and non-viral systems. Liposome-mediated nucleic acid drug delivery is the main method of non-viral delivery systems.

在基因治療應用中，脂質奈米顆粒已經被證明可作為核酸藥物的遞送載體。由陽離子脂質和其他共脂質諸如膽固醇、磷脂和PEG化脂質形成的脂質奈米顆粒包封核酸，保護核酸免於降解並且促進細胞攝取。另外，脂質體奈米顆粒用於生物活性成分的遞送具有其他優點，如靶向性好、副作用小、穩定性好和轉染效率較高等。In gene therapy applications, lipid nanoparticles have been demonstrated as delivery vehicles for nucleic acid drugs. Lipid nanoparticles formed from cationic lipids and other co-lipids such as cholesterol, phospholipids, and PEGylated lipids encapsulate nucleic acids, protecting them from degradation and facilitating cellular uptake. In addition, the delivery of liposomal nanoparticles for bioactive components has other advantages, such as good targeting, less side effects, good stability, and high transfection efficiency.

隨著基於mRNA的療法，例如疫苗、基因治療、蛋白替代療法等領域快速發展，對於mRNA的遞送系統提出了巨大的需求，因此開發高效、安全的mRNA遞送系統對基於mRNA等核酸藥物疾病，包括預防性疾病、遺傳性疾病和腫瘤等疾病的治療具有重要意義。With the rapid development of mRNA-based therapies, such as vaccines, gene therapy, and protein replacement therapy, there is a huge demand for mRNA delivery systems. Therefore, the development of efficient and safe mRNA delivery systems is effective for diseases based on mRNA and other nucleic acid drugs, including The treatment of diseases such as preventive diseases, genetic diseases and tumors is of great significance.

本揭露（The disclosure）提供了式I所示化合物或其鹽

其中，M ¹至M ⁶各自獨立地選自鍵、-C(O)O-a1和-OC(O)-a1，a1為與R ¹、R ²、R ³、R ⁴、R ⁵或R ⁶連接的鍵，且M ¹至M ⁶不全為-C(O)O-a1或鍵； R ¹至R ⁶各自獨立地為取代或未被取代的烷基或取代或未被取代的烯基； R ⁷各自獨立地為氫或取代或未被取代的C _1-6烷基； m1至m3各自獨立地選自1、2、3、4、5、6、7和8； n1至n6各自獨立地選自0、1、2、3、4、5、6、7、8、9和10。 The disclosure (The disclosure) provides a compound represented by formula I or a salt thereof

Wherein, M ¹ to M ⁶ are each independently selected from a bond, -C(O)O-a1 and -OC(O)-a1, and a1 is a combination with R ¹ , R ² , R ³ , R ⁴ , R ⁵ or R ⁶ , and M ¹ to M ⁶ are not all -C(O)O-a1 or bonds; R ¹ to R ⁶ are each independently substituted or unsubstituted alkyl or substituted or unsubstituted alkenyl ; R ⁷ each independently hydrogen or substituted or unsubstituted C _1-6 alkyl; m1 to m3 each independently selected from 1, 2, 3, 4, 5, 6, 7 and 8; n1 to n6 each independently selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10.

在一些實施方案中，式I所示化合物或其鹽中R ¹至R ⁶各自獨立地為未被取代的烷基或未被取代的烯基。 In some embodiments, in the compound represented by formula I or a salt thereof, R ¹ to R ⁶ are each independently unsubstituted alkyl or unsubstituted alkenyl.

在一些實施方案中，式I所示化合物或其鹽中R ¹至R ⁶各自獨立地選自C ₄-C ₁₄烷基或C ₄-C ₁₄烯基。 In some embodiments, in the compound represented by formula I or a salt thereof, R ¹ to R ⁶ are each independently selected from C ₄ -C ₁₄ alkyl or C ₄ -C ₁₄ alkenyl.

在一些實施方案中，式I所示化合物或其鹽中R ¹為C ₄-C ₁₄烷基（包括但不限於C ₄烷基、C ₅烷基、C ₆烷基、C ₇烷基、C ₈烷基、C ₉烷基、C ₁₀烷基、C ₁₁烷基、C ₁₂烷基、C ₁₃烷基、C ₁₄烷基）。在另一些實施方案中，式I所示化合物或其鹽中R ¹為C ₅-C ₁₂烷基。 In some embodiments, in the compound represented by formula I or its salt, R ¹ is C ₄ -C ₁₄ alkyl (including but not limited to C ₄ alkyl, C ₅ alkyl, C ₆ alkyl, C ₇ alkyl, C ₈ alkyl, C ₉ alkyl, C ₁₀ alkyl, C ₁₁ alkyl, C ₁₂ alkyl, C ₁₃ alkyl, C ₁₄ alkyl). In other embodiments, in the compound represented by formula I or a salt thereof, R ¹ is C ₅ -C ₁₂ alkyl.

在一些實施方案中，式I所示化合物或其鹽中R ²為C ₄-C ₁₄烷基（包括但不限於C ₄烷基、C ₅烷基、C ₆烷基、C ₇烷基、C ₈烷基、C ₉烷基、C ₁₀烷基、C ₁₁烷基、C ₁₂烷基、C ₁₃烷基、C ₁₄烷基）。 In some embodiments, in the compound represented by formula I or its salt, R ² is C ₄ -C ₁₄ alkyl (including but not limited to C ₄ alkyl, C ₅ alkyl, C ₆ alkyl, C ₇ alkyl, C ₈ alkyl, C ₉ alkyl, C ₁₀ alkyl, C ₁₁ alkyl, C ₁₂ alkyl, C ₁₃ alkyl, C ₁₄ alkyl).

在另一些實施方案中，式I所示化合物或其鹽中R ²為C ₅-C ₁₂烷基。 In other embodiments, in the compound represented by formula I or a salt thereof, R ² is a C ₅ -C ₁₂ alkyl group.

在一些實施方案中，式I所示化合物或其鹽中R ³為C ₄-C ₁₄烷基（包括但不限於C ₄烷基、C ₅烷基、C ₆烷基、C ₇烷基、C ₈烷基、C ₉烷基、C ₁₀烷基、C ₁₁烷基、C ₁₂烷基、C ₁₃烷基、C ₁₄烷基）。在另一些實施方案中，式I所示化合物或其鹽中R ³為C ₅-C ₁₂烷基。 In some embodiments, in the compound represented by formula I or its salt, R ³ is C ₄ -C ₁₄ alkyl (including but not limited to C ₄ alkyl, C ₅ alkyl, C ₆ alkyl, C ₇ alkyl, C ₈ alkyl, C ₉ alkyl, C ₁₀ alkyl, C ₁₁ alkyl, C ₁₂ alkyl, C ₁₃ alkyl, C ₁₄ alkyl). In other embodiments, in the compound represented by formula I or a salt thereof, R ³ is a C ₅ -C ₁₂ alkyl group.

在一些實施方案中，式I所示化合物或其鹽中R ⁴為C ₄-C ₁₄烷基（包括但不限於C ₄烷基、C ₅烷基、C ₆烷基、C ₇烷基、C ₈烷基、C ₉烷基、C ₁₀烷基、C ₁₁烷基、C ₁₂烷基、C ₁₃烷基、C ₁₄烷基）。 In some embodiments, in the compound represented by formula I or its salt, R ⁴ is C ₄ -C ₁₄ alkyl (including but not limited to C ₄ alkyl, C ₅ alkyl, C ₆ alkyl, C ₇ alkyl, C ₈ alkyl, C ₉ alkyl, C ₁₀ alkyl, C ₁₁ alkyl, C ₁₂ alkyl, C ₁₃ alkyl, C ₁₄ alkyl).

在另一些實施方案中，式I所示化合物或其鹽中R ⁴為C ₅-C ₁₂烷基。 In other embodiments, in the compound represented by formula I or a salt thereof, R ⁴ is a C ₅ -C ₁₂ alkyl group.

在一些實施方案中，式I所示化合物或其鹽中R ⁵為C ₄-C ₁₄烷基（包括但不限於C ₄烷基、C ₅烷基、C ₆烷基、C ₇烷基、C ₈烷基、C ₉烷基、C ₁₀烷基、C ₁₁烷基、C ₁₂烷基、C ₁₃烷基、C ₁₄烷基）。在另一些實施方案中，式I所示化合物或其鹽中R ⁵為C ₅-C ₁₂烷基。 In some embodiments, in the compound represented by formula I or its salt, R ⁵ is C ₄ -C ₁₄ alkyl (including but not limited to C ₄ alkyl, C ₅ alkyl, C ₆ alkyl, C ₇ alkyl, C ₈ alkyl, C ₉ alkyl, C ₁₀ alkyl, C ₁₁ alkyl, C ₁₂ alkyl, C ₁₃ alkyl, C ₁₄ alkyl). In other embodiments, in the compound represented by formula I or its salt, R ⁵ is C ₅ -C ₁₂ alkyl.

在一些實施方案中，式I所示化合物或其鹽中R ⁶為C ₄-C ₁₄烷基（包括但不限於C ₄烷基、C ₅烷基、C ₆烷基、C ₇烷基、C ₈烷基、C ₉烷基、C ₁₀烷基、C ₁₁烷基、C ₁₂烷基、C ₁₃烷基、C ₁₄烷基）。在另一些實施方案中，式I所示化合物或其鹽中R ⁶為C ₅-C ₁₂烷基。 In some embodiments, in the compound represented by formula I or its salt, R ⁶ is C ₄ -C ₁₄ alkyl (including but not limited to C ₄ alkyl, C ₅ alkyl, C ₆ alkyl, C ₇ alkyl, C ₈ alkyl, C ₉ alkyl, C ₁₀ alkyl, C ₁₁ alkyl, C ₁₂ alkyl, C ₁₃ alkyl, C ₁₄ alkyl). In other embodiments, in the compound represented by formula I or a salt thereof, R ⁶ is a C ₅ -C ₁₂ alkyl group.

另一些實施方案中，式I所示化合物或其鹽中R ¹為C ₄-C ₁₄烯基（包括但不限於C ₄烯基、C ₅烯基、C ₆烯基、C ₇烯基、C ₈烯基、C ₉烯基、C ₁₀烯基、C ₁₁烯基、C ₁₂烯基、C ₁₃烯基、C ₁₄烯基）。在另一些實施方案中，式I所示化合物或其鹽中R ¹為C ₅-C ₁₂烯基。 In other embodiments, in the compound represented by formula I or its salt, R ¹ is C ₄ -C ₁₄ alkenyl (including but not limited to C ₄ alkenyl, C ₅ alkenyl, C ₆ alkenyl, C ₇ alkenyl, C ₈ alkenyl, C ₉ alkenyl, C ₁₀ alkenyl, C _{11 alkenyl, C 12} _alkenyl , C ₁₃ alkenyl, C ₁₄ alkenyl). In other embodiments, in the compound represented by formula I or a salt thereof, R ¹ is C ₅ -C ₁₂ alkenyl.

在一些實施方案中，式I所示化合物或其鹽中R ²為C ₄-C ₁₄烯基（包括但不限於C ₄烯基、C ₅烯基、C ₆烯基、C ₇烯基、C ₈烯基、C ₉烯基、C ₁₀烯基、C ₁₁烯基、C ₁₂烯基、C ₁₃烯基、C ₁₄烯基）。在另一些實施方案中，式I所示化合物或其鹽中R ²為C ₅-C ₁₂烯基。 In some embodiments, in the compound represented by formula I or its salt, R ² is C ₄ -C ₁₄ alkenyl (including but not limited to C ₄ alkenyl, C ₅ alkenyl, C ₆ alkenyl, C ₇ alkenyl, C ₈ alkenyl, C ₉ alkenyl, C ₁₀ alkenyl, C _{11 alkenyl, C 12} _alkenyl , C ₁₃ alkenyl, C ₁₄ alkenyl). In other embodiments, in the compound represented by formula I or a salt thereof, R ² is C ₅ -C ₁₂ alkenyl.

在一些實施方案中，式I所示化合物或其鹽中R ³為C ₄-C ₁₄烯基（包括但不限於C ₄烯基、C ₅烯基、C ₆烯基、C ₇烯基、C ₈烯基、C ₉烯基、C ₁₀烯基、C ₁₁烯基、C ₁₂烯基、C ₁₃烯基、C ₁₄烯基）。在另一些實施方案中，式I所示化合物或其鹽中R ³為C ₅-C ₁₂烯基。 In some embodiments, in the compound represented by formula I or its salt, R ³ is C ₄ -C ₁₄ alkenyl (including but not limited to C ₄ alkenyl, C ₅ alkenyl, C ₆ alkenyl, C ₇ alkenyl, C ₈ alkenyl, C ₉ alkenyl, C ₁₀ alkenyl, C ₁₁ alkenyl, C ₁₂ alkenyl, C ₁₃ alkenyl, C ₁₄ alkenyl). In other embodiments, in the compound represented by formula I or a salt thereof, R ³ is C ₅ -C ₁₂ alkenyl.

在一些實施方案中，式I所示化合物或其鹽中R ⁴為C ₄-C ₁₄烯基（包括但不限於C ₄烯基、C ₅烯基、C ₆烯基、C ₇烯基、C ₈烯基、C ₉烯基、C ₁₀烯基、C ₁₁烯基、C ₁₂烯基、C ₁₃烯基、C ₁₄烯基）。在另一些實施方案中，式I所示化合物或其鹽中R ⁴為C ₅-C ₁₂烯基。 In some embodiments, in the compound represented by formula I or its salt, R ⁴ is C ₄ -C ₁₄ alkenyl (including but not limited to C ₄ alkenyl, C ₅ alkenyl, C ₆ alkenyl, C ₇ alkenyl, C ₈ alkenyl, C ₉ alkenyl, C ₁₀ alkenyl, C ₁₁ alkenyl, C ₁₂ alkenyl, C ₁₃ alkenyl, C ₁₄ alkenyl). In other embodiments, in the compound represented by formula I or a salt thereof, R ⁴ is C ₅ -C ₁₂ alkenyl.

在一些實施方案中，式I所示化合物或其鹽中R ⁵為C ₄-C ₁₄烯基（包括但不限於C ₄烯基、C ₅烯基、C ₆烯基、C ₇烯基、C ₈烯基、C ₉烯基、C ₁₀烯基、C ₁₁烯基、C ₁₂烯基、C ₁₃烯基、C ₁₄烯基）。在另一些實施方案中，式I所示化合物或其鹽中R ⁵為C ₅-C ₁₂烯基。 In some embodiments, in the compound represented by formula I or its salt, R ⁵ is C ₄ -C ₁₄ alkenyl (including but not limited to C ₄ alkenyl, C ₅ alkenyl, C ₆ alkenyl, C ₇ alkenyl, C ₈ alkenyl, C ₉ alkenyl, C ₁₀ alkenyl, C _{11 alkenyl, C 12} _alkenyl , C ₁₃ alkenyl, C ₁₄ alkenyl). In other embodiments, in the compound represented by formula I or a salt thereof, R ⁵ is C ₅ -C ₁₂ alkenyl.

在一些實施方案中，式I所示化合物或其鹽中R ⁶為C ₄-C ₁₄烯基（包括但不限於C ₄烯基、C ₅烯基、C ₆烯基、C ₇烯基、C ₈烯基、C ₉烯基、C ₁₀烯基、C ₁₁烯基、C ₁₂烯基、C ₁₃烯基、C ₁₄烯基）。在另一些實施方案中，式I所示化合物或其鹽中R ⁶為C ₅-C ₁₂烯基。 In some embodiments, in the compound represented by formula I or its salt, R ⁶ is C ₄ -C ₁₄ alkenyl (including but not limited to C ₄ alkenyl, C ₅ alkenyl, C ₆ alkenyl, C ₇ alkenyl, C ₈ alkenyl, C ₉ alkenyl, C ₁₀ alkenyl, C ₁₁ alkenyl, C ₁₂ alkenyl, C ₁₃ alkenyl, C ₁₄ alkenyl). In other embodiments, in the compound represented by formula I or a salt thereof, R ⁶ is C ₅ -C ₁₂ alkenyl.

進一步地，在一些實施方案中，式I所示化合物或其鹽中R ⁵為C ₄-C ₁₄烯基，R ⁶為C ₄-C ₁₄烯基。 Further, in some embodiments, in the compound represented by formula I or a salt thereof, R ⁵ is C ₄ -C ₁₄ alkenyl, and R ⁶ is C ₄ -C ₁₄ alkenyl.

在一些實施方案中，式I所示化合物或其鹽中R ³為C ₄-C ₁₄烯基，R ⁴為C ₄-C ₁₄烯基。 In some embodiments, in the compound represented by formula I or a salt thereof, R ³ is C ₄ -C ₁₄ alkenyl, and R ⁴ is C ₄ -C ₁₄ alkenyl.

在一些實施方案中，式I所示化合物或其鹽中R ¹為C ₄-C ₁₄烯基，R ²為C ₄-C ₁₄烯基。 In some embodiments, in the compound represented by formula I or a salt thereof, R ¹ is C ₄ -C ₁₄ alkenyl, and R ² is C ₄ -C ₁₄ alkenyl.

進一步地，在一些實施方案中，式I所示化合物或其鹽中R ¹為C ₄-C ₁₄烷基，R ²為C ₄-C ₁₄烷基。 Further, in some embodiments, in the compound represented by formula I or a salt thereof, R ¹ is a C ₄ -C ₁₄ alkyl group, and R ² is a C ₄ -C ₁₄ alkyl group.

在一些實施方案中，式I所示化合物或其鹽中R ³為C ₄-C ₁₄烷基，R ⁴為C ₄-C ₁₄烷基。 In some embodiments, in the compound represented by formula I or a salt thereof, R ³ is C ₄ -C ₁₄ alkyl, and R ⁴ is C ₄ -C ₁₄ alkyl.

在一些實施方案中，式I所示化合物或其鹽中R ⁵為C ₄-C ₁₄烷基，R ⁶為C ₄-C ₁₄烷基。 In some embodiments, in the compound represented by formula I or a salt thereof, R ⁵ is a C ₄ -C ₁₄ alkyl group, and R ⁶ is a C ₄ -C ₁₄ alkyl group.

另一方面，一些實施方案提供式I所示化合物或其鹽中R ¹至R ⁶各自獨立為C _4-15烷基或烯基。另一些實施方案提供式I所示化合物或其鹽中R ¹至R ⁶各自獨立為C _8-16烷基或烯基。 On the other hand, some embodiments provide that in the compound represented by formula I or a salt thereof, R ¹ to R ⁶ are each independently C _4-15 alkyl or alkenyl. Other embodiments provide that in the compound represented by formula I or a salt thereof, R ¹ to R ⁶ are each independently C _8-16 alkyl or alkenyl.

在一些實施方案中，式I所示化合物或其鹽中R ¹為C _4-15烷基或C _8-16烷基。 In some embodiments, in the compound represented by formula I or a salt thereof, R ¹ is C _4-15 alkyl or C _8-16 alkyl.

在一些實施方案中，式I所示化合物或其鹽中R ²為C _4-15烷基或C _8-16烷基。 In some embodiments, in the compound represented by formula I or a salt thereof, R ² is C _4-15 alkyl or C _8-16 alkyl.

在一些實施方案中，式I所示化合物或其鹽中R ³為C _4-15烷基或C _8-16烷基。 In some embodiments, in the compound represented by formula I or a salt thereof, R ³ is C _4-15 alkyl or C _8-16 alkyl.

在一些實施方案中，式I所示化合物或其鹽中R ⁴為C _4-15烷基或C _8-16烷基。 In some embodiments, in the compound represented by formula I or its salt, R ⁴ is C _4-15 alkyl or C _8-16 alkyl.

在一些實施方案中，式I所示化合物或其鹽中R ⁵為C _4-15烷基或C _8-16烷基。 In some embodiments, in the compound represented by formula I or a salt thereof, R ⁵ is C _4-15 alkyl or C _8-16 alkyl.

在一些實施方案中，式I所示化合物或其鹽中R ⁶為C _4-15烷基或C _8-16烷基。 In some embodiments, in the compound represented by formula I or a salt thereof, R ⁶ is C _4-15 alkyl or C _8-16 alkyl.

在一些實施方案中，式I所示化合物或其鹽中R ¹至R ⁶為C _4-15烷基或C _8-16烷基。 In some embodiments, in the compound represented by formula I or a salt thereof, R ¹ to R ⁶ are C _4-15 alkyl or C _8-16 alkyl.

在一些實施方案中，式I所示化合物或其鹽中R ¹為C _4-15烯基或C _8-16烯基。 In some embodiments, in the compound represented by formula I or a salt thereof, R ¹ is C _4-15 alkenyl or C _8-16 alkenyl.

在一些實施方案中，式I所示化合物或其鹽中R ²為C _4-15烯基或C _8-16烯基。 In some embodiments, in the compound represented by formula I or a salt thereof, R ² is C _4-15 alkenyl or C _8-16 alkenyl.

在一些實施方案中，式I所示化合物或其鹽中R ³為C _4-15烯基或C _8-16烯基。 In some embodiments, in the compound represented by formula I or a salt thereof, R ³ is C _4-15 alkenyl or C _8-16 alkenyl.

在一些實施方案中，式I所示化合物或其鹽中R ⁴為C _4-15烯基或C _8-16烯基。 In some embodiments, in the compound represented by formula I or a salt thereof, R ⁴ is C _4-15 alkenyl or C _8-16 alkenyl.

在一些實施方案中，式I所示化合物或其鹽中R ⁵為C _4-15烯基或C _8-16烯基。 In some embodiments, in the compound represented by formula I or a salt thereof, R ⁵ is C _4-15 alkenyl or C _8-16 alkenyl.

在一些實施方案中，式I所示化合物或其鹽中R ⁶為C _4-15烯基或C _8-16烯基。 In some embodiments, in the compound represented by formula I or a salt thereof, R ⁶ is C _4-15 alkenyl or C _8-16 alkenyl.

在一些實施方案中，式I所示化合物或其鹽中R ¹至R ⁶為C _4-15烯基或C _8-16烯基。 In some embodiments, in the compound represented by formula I or a salt thereof, R ¹ to R ⁶ are C _4-15 alkenyl or C _8-16 alkenyl.

另一方面，一些實施方案提供的式I所示化合物或其鹽中R ⁷為氫或視需要取代的C _1-3烷基。在一些實施方案中，式I所示化合物或其鹽中R ⁷為氫、甲基或乙基。 On the other hand, in the compound represented by formula I or a salt thereof provided by some embodiments, R ⁷ is hydrogen or optionally substituted C _1-3 alkyl. In some embodiments, in the compound represented by formula I or a salt thereof, R ⁷ is hydrogen, methyl or ethyl.

在一些實施方案中，式I所示化合物或其鹽中M ¹至M ⁶各自獨立地為-OC(O)-a1，a1為與R ¹、R ²、R ³、R ⁴、R ⁵或R ⁶連接的鍵。 In some embodiments, M ¹ to M ⁶ in the compound represented by formula I or its salt are each independently -OC(O)-a1, and a1 is the combination of R ¹ , R ² , R ³ , R ⁴ , R ⁵ or R ⁶ is connected to the bond.

在另一些實施方案中，式I所示化合物或其鹽中M ¹、M ²各自獨立地為-C(O)O-a1，a1為與R ¹或R ²連接的鍵。 In other embodiments, M ¹ and M ² in the compound represented by formula I or a salt thereof are each independently -C(O)O-a1, and a1 is a bond connecting R ¹ or R ² .

在另一些實施方案中，式I所示化合物或其鹽中M ¹、M ²、M ³、M ⁴各自獨立地為-C(O)O-a1，a1為與R ¹、R ²、R ³或R ⁴連接的鍵。 In other embodiments, M ¹ , M ² , M ³ , and M ⁴ in the compound represented by formula I or its salt are each independently -C(O)O-a1, and a1 is the same as R ¹ , R ² , R ³ or R ⁴ bonded.

另一方面，在一些實施方案中，式I所示化合物或其鹽中M ¹、M ²、M ³和M ⁴為鍵。 On the other hand, in some embodiments, in the compound represented by formula I or a salt thereof, M ¹ , M ² , M ³ and M ⁴ are bonds.

在另一些實施方案中，式I所示化合物或其鹽中M ⁵和M ⁶為鍵。 In other embodiments, M ⁵ and M ⁶ are bonds in the compound represented by formula I or a salt thereof.

進一步地，一些實施方案提供的式I所示化合物或其鹽中m1至m3各自獨立地選自2、3和4。Further, m1 to m3 in the compound represented by formula I or the salt thereof provided by some embodiments are each independently selected from 2, 3 and 4.

在一些實施方案中，式I所示化合物或其鹽中R ¹至R ⁶獨立地為C ₄-C ₁₄烷基或C ₄-C ₁₄烯基。 In some embodiments, in the compound represented by formula I or a salt thereof, R ¹ to R ⁶ are independently C ₄ -C ₁₄ alkyl or C ₄ -C ₁₄ alkenyl.

在一些實施方案中，式I所示化合物或其鹽中R ¹或R ²獨立地選自

、

、

、

和

。 In some embodiments, in the compound shown in formula I or its salt, R ¹ or R ² are independently selected from

,

and

.

在一些實施方案中，式I所示化合物或其鹽中n1至n6各自獨立地選自1、2、3、4和5。In some embodiments, n1 to n6 in the compound represented by formula I or a salt thereof are each independently selected from 1, 2, 3, 4 and 5.

在一些實施方案中，式I所示化合物或其鹽中n1至n6各自獨立地為3。In some embodiments, n1 to n6 in the compound represented by formula I or a salt thereof are 3 independently.

在一些實施方案中，式I所示化合物或其鹽中n1至n6各自獨立地為4。In some embodiments, n1 to n6 in the compound represented by formula I or a salt thereof are 4 independently.

在一些實施方案中，式I所示化合物或其鹽中n1至n6各自獨立地為5。In some embodiments, n1 to n6 in the compound represented by formula I or a salt thereof are 5 independently.

在一些實施方案中，式I所示化合物或其鹽中n1至n6各自獨立地選自6、7、8、9和10。In some embodiments, in the compound represented by formula I or a salt thereof, n1 to n6 are each independently selected from 6, 7, 8, 9 and 10.

在一些實施方案中，式I所示化合物或其鹽中n1至n6各自獨立地為7。In some embodiments, n1 to n6 in the compound represented by formula I or a salt thereof are each independently 7.

在一些實施方案中，式I所示化合物或其鹽中n1至n6各自獨立地為8。In some embodiments, n1 to n6 in the compound represented by formula I or a salt thereof are each independently 8.

在一些實施方案中，式I所示化合物或其鹽中n1至n6各自獨立地為9。In some embodiments, n1 to n6 in the compound represented by formula I or a salt thereof are each independently 9.

另一方面，本揭露式I所示化合物或其鹽為式II所示化合物或其鹽，

，其中R ¹至R ⁷、m1至m3、n1至n6如式I化合物或其鹽中定義。 On the other hand, the compound represented by formula I or its salt in this disclosure is the compound represented by formula II or its salt,

, wherein R ¹ to R ⁷ , m1 to m3, n1 to n6 are as defined in the compound of formula I or a salt thereof.

在一些實施方案中，式I或式II所示化合物或其鹽中n1或n2各自獨立地選自6、7、8、9和10。In some embodiments, in the compound represented by formula I or formula II or salt thereof, n1 or n2 are each independently selected from 6, 7, 8, 9 and 10.

在一些實施方案中，式II所示化合物或其鹽中R ¹或R ²獨立地選自

、

和

。 In some embodiments, in the compound shown in formula II or its salt, R ¹ or R ² are independently selected from

,

and

.

在一些實施方案中，式I或II所示化合物或其鹽中R ⁷獨立地為氫。 In some embodiments, in the compound represented by formula I or II or a salt thereof, R ⁷ is independently hydrogen.

在一些實施方案中，式II所示化合物或其鹽中m1至m3獨立地選自2、3和4。另一方面，本揭露式I所示化合物或其鹽為式III所示化合物或其鹽

，其中R ¹至R ⁷、m1至m3、n1至n6如式I化合物或其鹽中定義。 In some embodiments, in the compound represented by formula II or a salt thereof, m1 to m3 are independently selected from 2, 3 and 4. On the other hand, the compound represented by formula I or its salt in this disclosure is the compound represented by formula III or its salt

在一些實施方案中，式III所示化合物或其鹽中R ⁷獨立地為氫。 In some embodiments, in the compound represented by formula III or a salt thereof, R ⁷ is independently hydrogen.

在一些實施方案中，式III所示化合物或其鹽中m1至m3獨立地選自2、3和4。In some embodiments, in the compound represented by formula III or a salt thereof, m1 to m3 are independently selected from 2, 3 and 4.

在一些實施方案中，式III所示化合物或其鹽中n1至n6各自獨立地選自6、7、8、9和10。In some embodiments, n1 to n6 in the compound represented by formula III or a salt thereof are each independently selected from 6, 7, 8, 9 and 10.

在一些實施方案中，式III所示化合物或其鹽中R ¹至R ⁶獨立地選自

、

和

。 In some embodiments, in the compound shown in formula III or its salt, R ¹ to R ⁶ are independently selected from

,

and

.

在一些實施方案中，式III所示化合物或其鹽中R ¹至R ⁶獨立地為

。 In some embodiments, in the compound shown in formula III or its salt, R ¹ to R ⁶ are independently

.

式I所示典型化合物或其可藥用鹽，包括但不限於：

、

、

、

、

、

、

、

、

、

。 Typical compounds shown in formula I or pharmaceutically acceptable salts thereof include but are not limited to:

,

.

本揭露還提供一種前述的化合物或其鹽的同位素取代物，較佳地，該同位素取代物為氘原子取代物。The present disclosure also provides an isotope substitution of the aforementioned compound or a salt thereof, preferably, the isotope substitution is a deuterium atom substitution.

本揭露中還提供一種脂質顆粒，其包含前述化合物或其鹽，或同位素取代物。進一步地，在一些實施方案中，該脂質顆粒進一步包含活性劑，該活性劑較佳免疫刺激性寡核苷酸、siRNA、反義寡核苷酸、mRNA、質粒。The present disclosure also provides a lipid particle comprising the aforementioned compound or a salt thereof, or an isotope substitution. Further, in some embodiments, the lipid particle further comprises an active agent, the active agent is preferably immunostimulatory oligonucleotide, siRNA, antisense oligonucleotide, mRNA, plasmid.

本揭露還提供醫藥組成物，其包含前述的脂質顆粒和藥學上可接受的賦形劑。在某些實施方案中，基於組成物的總重量，該醫藥組成物含有0.01%-99.99%的藥學上可接受的賦形劑。在某些實施方案中，該醫藥組成物含有0.1%-99.9%的藥學上可接受的賦形劑。在某些實施方案中，該醫藥組成物含有0.5%-99.5%的藥學上可接受的賦形劑。在某些實施方案中，該醫藥組成物含有1%-99%的藥學上可接受的賦形劑。在某些實施方案中，該醫藥組成物含有2%-98%的藥學上可接受的賦形劑。The present disclosure also provides a pharmaceutical composition, which comprises the aforementioned lipid particles and pharmaceutically acceptable excipients. In certain embodiments, the pharmaceutical composition contains 0.01%-99.99% of pharmaceutically acceptable excipients based on the total weight of the composition. In certain embodiments, the pharmaceutical composition contains 0.1%-99.9% of pharmaceutically acceptable excipients. In certain embodiments, the pharmaceutical composition contains 0.5%-99.5% of pharmaceutically acceptable excipients. In certain embodiments, the pharmaceutical composition contains 1%-99% pharmaceutically acceptable excipients. In certain embodiments, the pharmaceutical composition contains 2%-98% pharmaceutically acceptable excipients.

本揭露還提供一種前述的化合物或其鹽，或同位素取代物，或前述脂質顆粒，或前述醫藥組成物在製備用於誘導受試者免疫反應的藥物中的用途。The present disclosure also provides a use of the aforementioned compound or its salt, or isotope substitution, or the aforementioned lipid particle, or the aforementioned pharmaceutical composition in the preparation of a medicament for inducing an immune response in a subject.

本揭露還提供一種前述的化合物或其鹽，或同位素取代物，或前述脂質顆粒，或前述醫藥組成物在製備用於預防和/或治療與多肽過表達相關的疾病或病症的藥物中的用途。The present disclosure also provides the use of the aforementioned compound or its salt, or isotope substitution, or the aforementioned lipid particle, or the aforementioned pharmaceutical composition in the preparation of medicines for preventing and/or treating diseases or disorders related to polypeptide overexpression .

本揭露還提供一種前述的化合物或其鹽，或同位素取代物，或前述脂質顆粒，或前述醫藥組成物在製備用於預防和/或治療與多肽表達不足相關的疾病或病症的藥物中的用途。The present disclosure also provides the use of the aforementioned compound or its salt, or isotope substitution, or the aforementioned lipid particle, or the aforementioned pharmaceutical composition in the preparation of medicines for preventing and/or treating diseases or conditions related to insufficient expression of polypeptides .

在一些實施方案中，本揭露所述疾病或病症包括但不限於：癌症、感染、自身免疫疾病、神經退行性疾病和炎症，例如新冠。In some embodiments, diseases or conditions described in the present disclosure include, but are not limited to, cancer, infection, autoimmune disease, neurodegenerative disease, and inflammation, such as COVID-19.

本揭露還提供一種用於誘導受試者免疫反應的方法，包括向該患者施用含有前述的化合物或其鹽，或前述脂質顆粒，或前述醫藥組成物。The present disclosure also provides a method for inducing an immune response in a subject, comprising administering to the patient a composition containing the aforementioned compound or a salt thereof, or the aforementioned lipid particle, or the aforementioned pharmaceutical composition.

本揭露還提供一種預防和/或治療與多肽過表達相關的疾病或病症的方法，包括向該患者施用含有前述的化合物或其鹽，或前述脂質顆粒，或前述醫藥組成物。The present disclosure also provides a method for preventing and/or treating diseases or diseases related to polypeptide overexpression, comprising administering to the patient a composition containing the aforementioned compound or its salt, or the aforementioned lipid particle, or the aforementioned pharmaceutical composition.

本揭露還提供一種預防和/或治療與多肽表達不足相關的疾病或病症的方法，包括向該患者施用含有前述的化合物或其鹽，或前述脂質顆粒，或前述醫藥組成物。The present disclosure also provides a method for preventing and/or treating a disease or condition related to insufficient expression of a polypeptide, comprising administering to the patient a composition containing the aforementioned compound or its salt, or the aforementioned lipid particle, or the aforementioned pharmaceutical composition.

另一方面，本揭露還提供用於誘導受試者免疫反應的前述的化合物或其鹽，或前述脂質顆粒，或前述醫藥組成物。On the other hand, the present disclosure also provides the aforementioned compound or its salt, or the aforementioned lipid particle, or the aforementioned pharmaceutical composition for inducing an immune response in a subject.

本揭露還提供用於預防和/或治療與多肽過表達相關的疾病或病症的前述的化合物或其鹽，或前述脂質顆粒，或前述醫藥組成物。The present disclosure also provides the aforementioned compound or its salt, or the aforementioned lipid particle, or the aforementioned pharmaceutical composition for preventing and/or treating diseases or conditions related to polypeptide overexpression.

本揭露還提供用於預防和/或治療與多肽表達不足相關的疾病或病症的前述的化合物或其鹽，或前述脂質顆粒，或前述醫藥組成物。The present disclosure also provides the aforementioned compound or its salt, or the aforementioned lipid particle, or the aforementioned pharmaceutical composition for preventing and/or treating diseases or conditions related to insufficient expression of polypeptides.

本揭露中所述化合物鹽包括“酸”加成鹽和“鹼”加成鹽。例如，藉由與鹼性基團（胺基）的酸-鹼反應形成的鹽，該酸包括有機酸或無機酸。另外，該化合物鹽還包括藉由與鹼性基團（胺基）的季胺化形成的鹽，該季胺化試劑包括直鏈或支鏈式的氯烴。Salts of compounds described in this disclosure include "acid" addition salts and "base" addition salts. For example, salts formed by acid-base reactions with basic groups (amine groups), including organic or inorganic acids. In addition, the salt of the compound also includes a salt formed by quaternization with a basic group (amine group), and the quaternization agent includes straight-chain or branched-chain chlorohydrocarbons.

本揭露化合物可以存在特定的幾何或立體異構體形式。本揭露設想所有的這類化合物，包括順式和反式異構體、(-)-和(+)-對對映體、(R)-和(S)-對映體、非對映異構體、(D)-異構體、(L)-異構體，及其外消旋混合物和其他混合物，例如對映異構體或非對映體富集的混合物，所有這些混合物都屬於本揭露的範圍之內。烷基等取代基中可存在另外的不對稱碳原子。所有這些異構體以及它們的混合物，均包括在本揭露的範圍之內。本揭露的含有不對稱碳原子的化合物可以以光學活性純的形式或外消旋形式被分離出來。光學活性純的形式可以從外消旋混合物拆分，或藉由使用手性原料或手性試劑合成。Compounds of the present disclosure may exist in particular geometric or stereoisomeric forms. This disclosure contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers isomers, (D)-isomers, (L)-isomers, and their racemic and other mixtures, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of this disclosure. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers, as well as mixtures thereof, are included within the scope of the present disclosure. Compounds of the present disclosure containing asymmetric carbon atoms can be isolated in optically pure or racemic forms. Optically pure forms can be resolved from racemic mixtures or synthesized by using chiral starting materials or reagents.

可以藉由的手性合成或手性試劑或者其他常規技術製備光學活性的(R)-和(S)-異構體以及D和L異構體。如果想得到本揭露某化合物的一種對映體，可以藉由不對稱合成或者具有手性助劑的衍生作用來製備，其中將所得非對映體混合物分離，並且輔助基團裂開以提供純的所需對映異構體。或者，當分子中含有鹼性官能團(如胺基)或酸性官能團(如羧基)時，與適當的光學活性的酸或鹼形成非對映異構體的鹽，然後藉由本領域所公知的常規方法進行非對映異構體拆分，然後回收得到純的對映體。此外，對映異構體和非對映異構體的分離通常是藉由使用色譜法完成的，該色譜法採用手性固定相，並視需要地與化學衍生法相結合(例如由胺生成胺基甲酸鹽)。Optically active (R)- and (S)-isomers as well as D and L-isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the present disclosure is desired, it can be prepared by asymmetric synthesis or derivatization with chiral auxiliary agents, wherein the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (such as an amine group) or an acidic functional group (such as a carboxyl group), a diastereoisomeric salt is formed with an appropriate optically active acid or base, and then by conventional methods known in the art Methods The diastereoisomers were resolved and the pure enantiomers were recovered. Furthermore, the separation of enantiomers and diastereomers is usually accomplished by the use of chromatography using a chiral stationary phase, optionally combined with chemical derivatization methods (e.g. amines to amines formate).

本揭露所述化合物的化學結構中，鍵“

”表示未指定構型，即如果化學結構中存在手性異構體，鍵“

”可以為“

”或“

”，或者同時包含“

”和“

”兩種構型。本揭露所述化合物的化學結構中，鍵“

”並未指定構型，即可以為Z構型或E構型，或者同時包含兩種構型。 In the chemical structures of the compounds described in this disclosure, the bond "

"indicates that no configuration is specified, i.e. if chiral isomers exist in the chemical structure, the bond"

"can be"

"or"

, or both "

"and"

"Two configurations. In the chemical structures of the compounds described in this disclosure, the bond"

" does not specify the configuration, it can be the Z configuration or the E configuration, or both configurations.

本揭露的化合物和中間體還可以以不同的互變異構體形式存在，並且所有這樣的形式包含於本揭露的範圍內。術語“互變異構體”或“互變異構體形式”是指可經由低能壘互變的不同能量的結構異構體。例如，質子互變異構體(也稱為質子轉移互變異構體)包括經由質子遷移的互變，如酮-烯醇及亞胺-烯胺、內醯胺-內醯亞胺異構化。內醯胺-內醯亞胺平衡實例是在如下所示的A和B之間。

The compounds and intermediates of the present disclosure may also exist in different tautomeric forms, and all such forms are included within the scope of the present disclosure. The term "tautomer" or "tautomeric form" refers to structural isomers of different energies that can interconvert via a low energy barrier. For example, proton tautomers (also known as prototropic tautomers) include interconversions via migration of a proton, such as keto-enol and imine-enamine, lactam-lactimine isomerizations. An example of a lactam-lactimine equilibrium is between A and B shown below.

本揭露中的所有化合物可以被畫成A型或B型。所有的互變異構形式在本揭露的範圍內。化合物的命名不排除任何互變異構體。All compounds in this disclosure can be drawn as either Form A or Form B. All tautomeric forms are within the scope of the present disclosure. The naming of compounds does not exclude any tautomers.

本揭露還包括一些與本文中記載的那些相同的，但一個或多個原子被原子量或質量數不同於自然中通常發現的原子量或質量數的原子置換的同位素標記的本揭露化合物。可結合到本揭露化合物的同位素的實例包括氫、碳、氮、氧、磷、硫、氟、碘和氯的同位素，諸如分別為 ²H、 ³H、 ¹¹C、 ¹³C、 ¹⁴C、 ¹³N、 ¹⁵N、 ¹⁵O、 ¹⁷O、 ¹⁸O、 ³¹P、 ³²P、 ³⁵S、 ¹⁸F、 ¹²³I、 ¹²⁵I和 ³⁶Cl等。 The disclosure also includes isotopically labeled compounds of the disclosure that are identical to those described herein, but wherein one or more atoms are replaced by an atom of an atomic mass or mass number different from that normally found in nature. Examples of isotopes that can be incorporated into compounds of the present disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, iodine, and chlorine, such as ² H, ³ H, ¹¹ C, ¹³ C, ¹⁴ C, ¹³ N, ¹⁵ N, ¹⁵ O, ¹⁷ O, ¹⁸ O, ³¹ P, ³² P, ³⁵ S, ¹⁸ F, ¹²³ I, ¹²⁵ I and ³⁶ Cl, etc.

除另有說明，當一個位置被特別地指定為氘（D）時，該位置應理解為具有大於氘的天然豐度（其為0.015%）至少1000倍的豐度的氘（即，至少10%的氘摻入）。示例中化合物的具有大於氘的天然豐度可以是至少1000倍的豐度的氘、至少2000倍的豐度的氘、至少3000倍的豐度的氘、至少4000倍的豐度的氘、至少5000倍的豐度的氘、至少6000倍的豐度的氘或更高豐度的氘。本揭露還包括各種氘化形式的式( I )化合物。與碳原子連接的各個可用的氫原子可獨立地被氘原子替換。所屬技術領域具有通常知識者能夠參考相關文獻合成氘化形式的式( I )化合物。在製備氘代形式的式( I )化合物時可使用市售的氘代起始物質，或它們可使用常規技術採用氘代試劑合成，氘代試劑包括但不限於氘代硼烷、三氘代硼烷四氫呋喃溶液、氘代氫化鋰鋁、氘代碘乙烷和氘代碘甲烷等。Unless otherwise stated, when a position is specifically designated as deuterium (D), the position is understood to have an abundance of deuterium (i.e., at least 10 % deuterium incorporation). Exemplary compounds having a natural abundance greater than deuterium can be at least 1000 times more abundant deuterium, at least 2000 times more abundant deuterium, at least 3000 times more abundant deuterium, at least 4000 times more abundant deuterium, at least 5000 times more abundant deuterium, at least 6000 times more abundant deuterium, or more abundant deuterium. The present disclosure also includes compounds of formula (I) in various deuterated forms. Each available hydrogen atom attached to a carbon atom can be independently replaced by a deuterium atom. Those skilled in the art can refer to the relevant literature to synthesize the deuterated form of the compound of formula (I). Commercially available deuterated starting materials can be used in the preparation of deuterated forms of compounds of formula (I), or they can be synthesized using conventional techniques using deuterated reagents, including but not limited to deuterated borane, trideuterated Borane tetrahydrofuran solution, deuterated lithium aluminum hydride, deuterated ethyl iodide and deuterated methyl iodide, etc.

“視需要地”或“視需要”是指意味著隨後所描述的事件或環境可以但不必發生，該說明包括該事件或環境發生或不發生的場合。例如“視需要的被鹵素或者氰基取代的C _1-6烷基”是指鹵素或者氰基可以但不必須存在，該說明包括烷基被鹵素或者氰基取代的情形和烷基不被鹵素和氰基取代的情形。 "Optionally" or "optionally" means that the subsequently described event or circumstance can but need not occur, and that the description includes instances where the event or circumstance occurs or does not occur. For example, "C _1-6 alkyl optionally substituted by halogen or cyano" means that halogen or cyano may but not necessarily exist, and this description includes the case where the alkyl is substituted by halogen or cyano and the alkyl is not substituted by halogen. And the case of cyano substitution.

術語解釋： “醫藥組成物”表示含有一種或多種本文所述化合物或其生理學上可藥用鹽或前體藥物與其他化學組分的混合物，以及其他組分例如生理學可藥用的載體和賦形劑。醫藥組成物的目的是促進對生物體的給藥，利於活性成分的吸收進而發揮生物活性。 Explanation of terms: "Pharmaceutical composition" means a mixture containing one or more compounds described herein or their physiologically acceptable salts or prodrugs and other chemical components, as well as other components such as physiologically acceptable carriers and excipients. agent. The purpose of the pharmaceutical composition is to promote the administration to the living body, facilitate the absorption of the active ingredient and thus exert the biological activity.

“可藥用賦形劑”包括但不限於任何已經被美國食品和藥物管理局批准對於人類或家畜動物使用可接受的任何助劑、載體、賦形劑、助流劑、甜味劑、稀釋劑、防腐劑、染料/著色劑、增香劑、表面活性劑、潤濕劑、分散劑、助懸劑、穩定劑、等滲劑、溶劑或乳化劑。"Pharmaceutically acceptable excipients" include, but are not limited to, any adjuvants, carriers, excipients, glidants, sweeteners, diluents, agent, preservative, dye/colorant, flavor enhancer, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, solvent or emulsifier.

本揭露中所述“有效量”或“有效治療量”包含足以改善或預防醫學病症的症狀或病症的量。有效量還意指足以允許或促進診斷的量。用於特定患者或獸醫學受試者的有效量可依據以下因素而變化：如待治療的病症、患者的總體健康情況、給藥的方法途徑和劑量以及副作用嚴重性。有效量可以是避免顯著副作用或毒性作用的最大劑量或給藥方案。An "effective amount" or "therapeutically effective amount" as used in this disclosure includes an amount sufficient to ameliorate or prevent a symptom or condition of a medical condition. An effective amount also means an amount sufficient to allow or facilitate diagnosis. Effective amounts for a particular patient or veterinary subject may vary depending on factors such as the condition being treated, the general health of the patient, the method, route and dosage of administration, and the severity of side effects. An effective amount may be the maximum dose or dosing regimen that avoids significant side effects or toxic effects.

術語“核酸”、“核酸分子”、“寡核苷酸”、“多核苷酸”和“核苷酸”在本文中可以可互換地使用。該術語關於脫氧核糖核苷酸(DNA)、核糖核苷酸(RNA)及其修飾的形式的聚合物，該聚合物呈單獨的片段或作為較大構建體的組分、直鏈或支鏈的、單鏈、雙鏈、三鏈或其雜合體的形式。該術語還包括RNA/DNA雜合體。多核苷酸可以包括DNA或RNA的有義寡核苷酸和反義寡核苷酸(sense and antisense oligonucleotide)或多核苷酸序列。DNA或RNA分子可以是，例如，但不限於：互補DNA(cDNA)、基因組DNA、合成的DNA、重組DNA或其雜合體，或RNA分子，諸如，例如，mRNA、shRNA、siRNA、miRNA、反義RNA和類似物。每種可能性代表本發明的單獨實施方案。該術語還包括包含天然存在的鹼基、糖和共價的核苷間鍵的寡核苷酸，以及具有非天然存在的部分的寡核苷酸，該非天然存在的部分的功能類似於相應的天然存在的部分。術語“多肽”、“肽”和“蛋白質”在本文中可互換地用於指胺基酸殘基的聚合物。該術語應用於其中一個或更多個胺基酸殘基是相應的天然存在的胺基酸的人工化學類似物的胺基酸聚合物，以及應用於天然存在的胺基酸聚合物。The terms "nucleic acid", "nucleic acid molecule", "oligonucleotide", "polynucleotide" and "nucleotide" are used interchangeably herein. The term pertains to polymers of deoxyribonucleotides (DNA), ribonucleotides (RNA) and modified forms thereof, either as individual fragments or as components of larger constructs, linear or branched single-stranded, double-stranded, triple-stranded or hybrid forms thereof. The term also includes RNA/DNA hybrids. Polynucleotides may include sense and antisense oligonucleotides or polynucleotide sequences of DNA or RNA. The DNA or RNA molecule can be, for example, but not limited to: complementary DNA (cDNA), genomic DNA, synthetic DNA, recombinant DNA or a hybrid thereof, or an RNA molecule such as, for example, mRNA, shRNA, siRNA, miRNA, anti- Sense RNA and the like. Each possibility represents a separate embodiment of the invention. The term also includes oligonucleotides comprising naturally occurring bases, sugars, and covalent internucleoside linkages, as well as oligonucleotides having non-naturally occurring portions that function similarly to the corresponding naturally occurring parts. The terms "polypeptide", "peptide" and "protein" are used interchangeably herein to refer to a polymer of amino acid residues. The term applies to amino acid polymers in which one or more amino acid residues are artificial chemical analogs of the corresponding naturally occurring amino acid, as well as to naturally occurring amino acid polymers.

術語“白細胞” 關於產生自和源自骨髓中的多能造血幹細胞的白細胞(WBC)。白細胞具有細胞核，並且基於功能特性或物理特性，白細胞的類型可以分為五種主要類型，包括中性粒細胞、嗜酸性粒細胞、嗜鹼性粒細胞、淋巴細胞和單核細胞。The term "leukocyte" refers to white blood cells (WBC) produced and derived from pluripotent hematopoietic stem cells in the bone marrow. White blood cells have a nucleus, and based on their functional or physical properties, the types of white blood cells can be divided into five main types, including neutrophils, eosinophils, basophils, lymphocytes, and monocytes.

“烷基”指飽和的脂族烴基團，包括但不限於直鏈和支鏈烷基基團。在一些實施方案中，烷基基團具有1-4個碳，也被稱為C _1-4烷基。在一些實施方案中，烷基基團具有10-22個碳，也被稱為C _10-22烷基。在一些實施方案中，烷基基團具有4-22個碳，也被稱為C _4-22烷基。在一些實施方案中，烷基基團具有4-15個碳，也被稱為C _4-15烷基。在一些實施方案中，烷基基團具有8-16個碳，也被稱為C _8-16烷基。 "Alkyl" refers to a saturated aliphatic hydrocarbon group including, but not limited to, straight and branched chain alkyl groups. In some embodiments, an alkyl group has 1-4 carbons, also known as C _1-4 alkyl. In some embodiments, an alkyl group has 10-22 carbons, also known as a C _10-22 alkyl. In some embodiments, an alkyl group has 4-22 carbons, also known as a _C4-22 alkyl. In some embodiments, an alkyl group has 4-15 carbons, also known as a C _4-15 alkyl. In some embodiments, the alkyl group has 8-16 carbons, also known as _C8-16 alkyl.

烷基基團可以是未被取代的或被一個或更多個選自鹵素、羥基、胺基、側氧、烷氧基羰基、醯胺基、烷基醯胺基、二烷基醯胺基、硝基、胺基、烷基胺基、二烷基胺基、羧基、硫基和硫烷基的基團取代。在一些實施方案中，烷基基團為直鏈烷基。在一些實施方案中，烷基基團為支鏈烷基。Alkyl groups may be unsubstituted or replaced by one or more members selected from halogen, hydroxyl, amine, pendant oxygen, alkoxycarbonyl, amido, alkylamido, dialkylamido , nitro, amine, alkylamine, dialkylamine, carboxyl, thio and sulfanyl groups. In some embodiments, the alkyl group is a straight chain alkyl. In some embodiments, the alkyl group is a branched chain alkyl.

“烯基”指不飽和的脂族烴基團，包括直鏈和支鏈烯基基團。在一些實施方案中，烯基基團具有1-4個碳，也被稱為C _1-4烯基。在一些實施方案中，烯基基團具有10-22個碳，也被稱為C _10-22烯基。在一些實施方案中，烯基基團具有4-22個碳，也被稱為C _4-22烯基。示例性的烯基基團包括乙烯基、丙烯基、正丁烯基、異丁烯基、3-甲基丁-2-烯基、正戊烯基、庚烯基、辛烯基、環己基-丁烯基和癸烯基。 "Alkenyl" means an unsaturated aliphatic hydrocarbon group and includes straight and branched chain alkenyl groups. In some embodiments, alkenyl groups have 1-4 carbons, also known as C _1-4 alkenyl. In some embodiments, alkenyl groups have 10-22 carbons, also known as C _10-22 alkenyl. In some embodiments, alkenyl groups have 4-22 carbons, also known as _C4-22 alkenyl. Exemplary alkenyl groups include vinyl, propenyl, n-butenyl, isobutenyl, 3-methylbut-2-enyl, n-pentenyl, heptenyl, octenyl, cyclohexyl-butenyl Alkenyl and Decenyl.

烯基可以是未被取代的或被一個或更多個選自鹵素、羥基、胺基、側氧、烷氧基羰基、醯胺基、烷基醯胺基、二烷基醯胺基、硝基、胺基、烷基胺基、二烷基胺基、羧基、硫基和硫烷基的基團取代。Alkenyl may be unsubstituted or replaced by one or more members selected from halogen, hydroxy, amine, pendant oxygen, alkoxycarbonyl, amido, alkylamido, dialkylamido, nitrate group, amine group, alkylamine group, dialkylamine group, carboxyl group, thio group and sulfanyl group substitution.

術語“羥基”指-OH基團。The term "hydroxyl" refers to a -OH group.

術語“鹵素”指氟、氯、溴或碘。The term "halogen" refers to fluorine, chlorine, bromine or iodine.

術語“氰基”指-NH ₂。 The term "cyano" refers to _-NH2 .

術語“側氧”指＝O取代基。The term "pendent oxygen" refers to a =O substituent.

“取代的”指基團中的一個或多個氫原子，較佳為最多5個，更佳為1～3個氫原子彼此獨立地被相應數目的取代基取代。不言而喻，取代基僅處在它們的可能的化學位置，所屬技術領域具有通常知識者能夠在不付出過多努力的情況下確定(藉由實驗或理論)可能或不可能的取代。"Substituted" means that one or more hydrogen atoms in a group, preferably up to 5, more preferably 1 to 3 hydrogen atoms are independently substituted by the corresponding number of substituents. It goes without saying that substituents are only in their possible chemical positions and that a person skilled in the art is able to determine (by experiment or theory) possible or impossible substitutions without undue effort.

以下結合實施例進一步描述本揭露中，但這些實施例並非限制本揭露中的範圍。The present disclosure is further described below in conjunction with examples, but these examples do not limit the scope of the present disclosure.

本揭露中實施例中未註明具體條件的實驗方法，通常按照常規條件，或按照原料或商品製造廠商所建議的條件。未註明具體來源的試劑，為市場購買的常規試劑。The experimental methods in the examples of the present disclosure that do not indicate specific conditions are usually in accordance with conventional conditions, or in accordance with the conditions suggested by the raw material or commodity manufacturers. Reagents without specific sources indicated are conventional reagents purchased in the market.

化合物的結構是藉由核磁共振(NMR)或/和質譜(MS)來確定的。NMR位移(δ)以10 ^-6(ppm)的單位給出。NMR的測定是用Bruker AVANCE-400核磁儀，測定溶劑為氘代二甲基亞碸(DMSO- d ₆ )，氘代氯仿(CDCl ₃)，氘代甲醇(Methanol- d ₄ )，內標為四甲基矽烷(TMS)。 Compound structures were determined by nuclear magnetic resonance (NMR) or/and mass spectroscopy (MS). NMR shifts (δ) are given in units of 10 ^-6 (ppm). The determination of NMR is to use Bruker AVANCE-400 nuclear magnetic instrument, and the measuring solvent is deuterated dimethyl sulfide (DMSO- d ₆ ), deuterated chloroform (CDCl ₃ ), deuterated methanol (Methanol- d ₄ ), and the internal standard is Tetramethylsilane (TMS).

HPLC的測定使用Agilent1100高壓液相色譜儀，GAS15B DAD紫外檢測器，Water Vbridge C18 150*4.6mm 5um色譜管柱。The determination of HPLC uses Agilent1100 high pressure liquid chromatography, GAS15B DAD ultraviolet detector, Water Vbridge C18 150*4.6mm 5um chromatographic column.

MS的測定用Agilent6120 三重四級杆質譜儀，G1315D DAD檢測器，Waters Xbridge C18 4.6*50mm,5um色譜管柱，以正/負離子模式掃描，質量掃描範圍為80~1200。The determination of MS uses Agilent6120 triple quadrupole mass spectrometer, G1315D DAD detector, Waters Xbridge C18 4.6*50mm, 5um chromatographic column, scanning in positive/negative ion mode, and the mass scanning range is 80~1200.

薄層層析矽膠板使用煙臺黃海HSGF254矽膠板，薄層色譜法(TLC)使用矽膠板採用規格是0.2mm±0.03mm，薄層層析分離純化產品採用的規格是0.4mm-0.5 mm。The thin-layer chromatography silica gel plate uses Yantai Huanghai HSGF254 silica gel plate, the specification of the thin-layer chromatography (TLC) silica gel plate is 0.2mm±0.03mm, and the specification of the thin-layer chromatography separation and purification product is 0.4mm-0.5mm.

快速管柱純化系統使用Combiflash Rf150 (TELEDYNE ISCO)或者Isolara one（Biotage）。The flash column purification system uses Combiflash Rf150 (TELEDYNE ISCO) or Isolara one (Biotage).

正向管柱層析一般使用煙臺黃海矽膠200~300目或300~400目矽膠為載體，或者使用常州三泰預填預填超純正相矽膠管柱（40-63μm，60g，24g，40g，120g或其它規格）。Forward column chromatography generally uses Yantai Huanghai silica gel 200~300 mesh or 300~400 mesh silica gel as the carrier, or Changzhou Santai prepacked prepacked ultrapure normal phase silica gel column (40-63μm, 60g, 24g, 40g, 120g or other specifications).

本揭露中的已知的起始原料可以採用或按照本領域已知的方法來合成，或可購買自上海泰坦科技，ABCR GmbH & Co. KG，Acros Organics，Aldrich Chemical Company，韶遠化學科技(Accela ChemBio Inc)，畢得醫藥等公司。The known starting materials in this disclosure can be used or synthesized according to methods known in the art, or can be purchased from Shanghai Titan Technology, ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Shaoyuan Chemical Technology ( Accela ChemBio Inc), Bi De Pharmaceutical and other companies.

實施例中無特殊說明，反應能夠均在氮氣氛下進行。Unless otherwise specified in the examples, the reactions can all be carried out under a nitrogen atmosphere.

氮氣氛是指反應瓶連接一個約1L容積的氮氣氣球。The nitrogen atmosphere means that the reaction bottle is connected to a nitrogen balloon with a volume of about 1 L.

氫氣氛是指反應瓶連接一個約1L容積的氫氣氣球。The hydrogen atmosphere means that the reaction bottle is connected to a hydrogen balloon with a capacity of about 1L.

氫氣是由上海全浦科學儀器公司QPH-1L型氫氣發生儀製得。Hydrogen was produced by a QPH-1L hydrogen generator from Shanghai Quanpu Scientific Instrument Company.

氮氣氛或氫化氛通常抽真空，充入氮氣或氫氣，重複操作3次。The nitrogen atmosphere or hydrogenation atmosphere is usually evacuated, filled with nitrogen or hydrogen, and the operation is repeated 3 times.

實施例中無特殊說明，溶液是指水溶液。Unless otherwise specified in the examples, the solution refers to an aqueous solution.

實施例中無特殊說明，反應的溫度為室溫，為20℃～30℃。Unless otherwise specified in the examples, the reaction temperature is room temperature, which is 20°C to 30°C.

實施例中的反應進程的監測採用薄層色譜法(TLC)，反應所使用的展開劑，純化化合物採用的管柱層析的沖提劑的體系和薄層色譜法的展開劑體系，溶劑的體積比根據化合物的極性不同而進行調節，也可以加入少量的三乙胺和醋酸等鹼性或酸性試劑進行調節。The monitoring of the reaction process in the embodiment adopts thin-layer chromatography (TLC), the developing agent used in the reaction, the eluting agent system and the developing agent system of the column chromatography that the purified compound adopts and the thin-layer chromatography, the solvent The volume ratio is adjusted according to the polarity of the compound, and can also be adjusted by adding a small amount of basic or acidic reagents such as triethylamine and acetic acid.

實施例1 二(辛烷-4-基) 9,9'-((3-(3,5-雙((3-(二(十二烷基胺基))丙基)胺基甲醯基)苯甲醯胺基)丙基)氮烷基二基)二壬酸酯

Example 1 Di(octane-4-yl) 9,9'-((3-(3,5-bis((3-(di(dodecylamino))propyl)aminoformyl ) benzamido) propyl) azaalkyl diyl) dinonanoate

化合物 E的製備

Preparation of Compound E

於氮氣保護下，將化合物 E1(13 g, 55 mmol)和化合物 E2(6.5 g, 50 mmol)溶解於DCM (65 mL)，加入DMAP (1.8 g, 15 mmol)和EDCI (14.4g, 75 mmol)，室溫下反應，TLC（EA/PE = 1/10）顯示反應達到終點，將反應液與矽膠（100-200目）拌樣濃縮，100-200目矽膠管柱層析EA/PE = 1/30 得14.14 g油狀物化合物 E3，收率：81.2%。 ¹H NMR (400 MHz,CDCl ₃)： δ4.92-4.83 (m, 1 H), 3.42-3.32 (m, 2 H), 2.30-2.23 (m, 2 H), 1.87-1.73 (m, 2 H), 1.63-1.18 (m, 20 H), 0.96-0.89(m, 6 H)。 Under nitrogen protection, compound E1 (13 g, 55 mmol) and compound E2 (6.5 g, 50 mmol) were dissolved in DCM (65 mL), DMAP (1.8 g, 15 mmol) and EDCI (14.4 g, 75 mmol) were added ), reacted at room temperature, TLC (EA/PE = 1/10) showed that the reaction reached the end point, the reaction solution was mixed with silica gel (100-200 mesh) and concentrated, 100-200 mesh silica gel column chromatography EA/PE = 1/30 14.14 g of oil compound E3 was obtained, yield: 81.2%. ¹ H NMR (400 MHz, CDCl ₃ ): δ 4.92-4.83 (m, 1 H), 3.42-3.32 (m, 2 H), 2.30-2.23 (m, 2 H), 1.87-1.73 (m, 2 H ), 1.63-1.18 (m, 20H), 0.96-0.89(m, 6H).

於氮氣保護下，將化合物 E3(9.0g, 25.8 mmol)和化合物 V2(1.8 g, 10.3 mmol)溶解於DMF (18 mL)中，加入K ₂CO ₃(3.56 g, 25.8 mmol)，80℃反應，TLC（MeOH/DCM = 1/10）檢測反應完全，乙酸乙酯（100 mL）稀釋，用水洗3次，硫酸鈉乾燥，過濾濃縮，矽膠管柱純化，MeOH/DCM體系，MeOH%：0%→15%，得到2.54g油狀物化合物 E4，產率34.6%。 ¹H NMR (400 MHz,CDCl ₃)： δ5.20 (bs, 1 H), 4.92-4.83 (m, 2 H), 3.47-3.32 (m, 2 H), 2.68-2.22 (m, 10 H), 1.87-1.39 (m, 46 H), 0.93-0.85(m, 12 H)。 Under the protection of nitrogen, compound E3 (9.0 g, 25.8 mmol) and compound V2 (1.8 g, 10.3 mmol) were dissolved in DMF (18 mL), and K ₂ CO ₃ (3.56 g, 25.8 mmol) was added to react at 80°C , TLC (MeOH/DCM = 1/10) detected that the reaction was complete, diluted with ethyl acetate (100 mL), washed 3 times with water, dried over sodium sulfate, concentrated by filtration, purified by silica gel column, MeOH/DCM system, MeOH%: 0 %→15%, to obtain 2.54g oil compound E4 , yield 34.6%. ¹ H NMR (400 MHz, CDCl ₃ ): δ 5.20 (bs, 1 H), 4.92-4.83 (m, 2 H), 3.47-3.32 (m, 2 H), 2.68-2.22 (m, 10 H), 1.87-1.39 (m, 46H), 0.93-0.85 (m, 12H).

將化合物 E4(2.54 g, 3.58 mmol)溶解於1,4-二噁烷 (10 mL)中，室溫下加入4M HCl (10 mL, 40.0 mmol)，室溫下反應2小時，LCMS顯示反應結束，濃縮至乾，得2.15 g粗品化合物 E，產率88.5%。 MS m/z (ESI): 611.5[M+1] ⁺。 Compound E4 (2.54 g, 3.58 mmol) was dissolved in 1,4-dioxane (10 mL), 4M HCl (10 mL, 40.0 mmol) was added at room temperature, and reacted at room temperature for 2 hours, LCMS showed that the reaction was complete , and concentrated to dryness to obtain 2.15 g of crude compound E with a yield of 88.5%. MS m/z (ESI): 611.5 [M+1] ⁺ .

化合物 V的製備

Preparation of compound V

於氮氣保護下，將化合物 V1(10.7 g, 42.9 mmol)和化合物 V3(3.0 g, 17.2 mmol)溶解於DMF (30 mL)中，加入碳酸鉀(5.95 g, 43.1 mmol)，80 ℃加熱反應過夜，反應液用乙酸乙酯稀釋，用水洗後，再用少量乙酸乙酯反萃水相，合併有機相，水洗3次，食鹽水洗一次，硫酸鈉乾燥，過濾濃縮，快速管柱層析純化，沖提體系MeOH/DCM，MeOH%：0%-5%(5min)→5%-10%(10 min)→10%-15%(15 min)得3.8克油狀物化合物 V2，產率43.2%。 ¹H NMR (400 MHz, CDCl ₃)：δ 5.74 (s, 1H), 3.25-3.09 (m, 2H), 2.55-2.25 (m, 6H), 1.69-1.54 (m, 2H), 1.50-1.16 (m, 49H), 0.88 (t, 6H)。 Under nitrogen protection, compound V1 (10.7 g, 42.9 mmol) and compound V3 (3.0 g, 17.2 mmol) were dissolved in DMF (30 mL), potassium carbonate (5.95 g, 43.1 mmol) was added, and the reaction was heated at 80 °C overnight , the reaction solution was diluted with ethyl acetate, washed with water, and the aqueous phase was back-extracted with a small amount of ethyl acetate, the organic phases were combined, washed 3 times with water, washed once with saline, dried over sodium sulfate, concentrated by filtration, and purified by flash column chromatography. Extraction system MeOH/DCM, MeOH%: 0%-5% (5min) → 5%-10% (10 min) → 10%-15% (15 min) to obtain 3.8 grams of oil compound V2 , yield 43.2 %. ¹ H NMR (400 MHz, CDCl ₃ ): δ 5.74 (s, 1H), 3.25-3.09 (m, 2H), 2.55-2.25 (m, 6H), 1.69-1.54 (m, 2H), 1.50-1.16 ( m, 49H), 0.88 (t, 6H).

將化合物 V2(2.0 g, 3.91 mmol)溶解於1,4-二噁烷 (18 mL)中，室溫下加入4M HCl (6 mL, 24.0 mmol)，50℃下反應1.0~3.0小時，濃縮至乾得1.92 g粗品化合物 V，所得粗品直接用於下一步。 MS m/z (ESI): 411.5 [M+1] ⁺ Dissolve compound V2 (2.0 g, 3.91 mmol) in 1,4-dioxane (18 mL), add 4M HCl (6 mL, 24.0 mmol) at room temperature, react at 50°C for 1.0-3.0 hours, and concentrate to Drying yielded 1.92 g of crude compound V , which was directly used in the next step. MS m/z (ESI): 411.5 [M+1] ⁺

步驟 1 ：化合物 1b的製備

Step 1 : Preparation of compound 1b

將化合物 V(800mg, 1.66 mmol)和化合物 1a(185mg, 0.83 mmol採用公知的方法“ Tetrahedron Letters, 2011, 52(1), 155 - 158”製備而得))溶解於DCM (8 mL)中,加入EDCI (478 mg, 2.49 mmol)，DMAP (101 mg, 0.83 mmol)和三乙胺(340 μL, 2.49 mmol)，反應於室溫下攪拌過夜，乙酸乙酯稀釋，水洗一次，硫酸鈉乾燥，過濾濃縮，快速管柱層析純化，沖提體系MeOH/DCM，MeOH%：0%-10%(10 min)→10%(5 min)→10%-20%(10 min)→20%-30%(10 min)得340 mg油狀物化合物 1b，產率40.8%。 ¹H NMR (400 MHz, CDCl ₃): δ 8.98 (s, 2H), 8.54 (s, 3H), 3.94 (s, 3H), 3.64-3.52 (m, 4H), 2.75-2.60 (m, 4H), 2.60-2.37 (m, 8H), 1.93-1.68 (m, 4H), 1.57-1.39 (m, 8H), 1.37-1.03 (m, 72H), 0.88 (t, 12H)。 Compound V (800 mg, 1.66 mmol) and compound 1a (185 mg, 0.83 mmol were prepared by the known method " Tetrahedron Letters , 2011, 52(1), 155-158")) were dissolved in DCM (8 mL), EDCI (478 mg, 2.49 mmol), DMAP (101 mg, 0.83 mmol) and triethylamine (340 μL, 2.49 mmol) were added, and the reaction was stirred overnight at room temperature, diluted with ethyl acetate, washed once with water, and dried over sodium sulfate. Concentrate by filtration, purify by flash column chromatography, extract system MeOH/DCM, MeOH%: 0%-10%(10 min)→10%(5 min)→10%-20%(10 min)→20%- 30% (10 min) to obtain 340 mg of oil compound 1b , yield 40.8%. ¹ H NMR (400 MHz, CDCl ₃ ): δ 8.98 (s, 2H), 8.54 (s, 3H), 3.94 (s, 3H), 3.64-3.52 (m, 4H), 2.75-2.60 (m, 4H) , 2.60-2.37 (m, 8H), 1.93-1.68 (m, 4H), 1.57-1.39 (m, 8H), 1.37-1.03 (m, 72H), 0.88 (t, 12H).

步驟 2 ：化合物 1c的製備

Step 2 : Preparation of Compound 1c

將化合物 1b(340mg, 0.337 mmol)溶解於THF/MeOH (1/1，8 mL)中，加入LiOH-H ₂O (33.6mg, 0.80 mmol)，於50℃下加熱反應完全，用稀鹽酸和飽和碳酸氫鈉溶液調節pH至7-8之間，並於室溫放置過夜，濃縮，用三乙胺調節pH至11左右，再用大板純化（MeOH/DCM = 1/5）得153g化合物 1c，產率45.7%。 ¹H NMR (400 MHz, CDCl ₃): δ 8.90 (s, 2H), 8.57 (s, 1H), 3.72-3.45 (m, 4H), 3.11-2.40 (m, 12H), 2.17-1.85 (m, 4H), 1.71-1.46 (m, 8H), 1.37-0.96 (m, 72H), 0.86 (t, 12H)。 Compound 1b (340mg, 0.337 mmol) was dissolved in THF/MeOH (1/1, 8 mL), LiOH-H ₂ O (33.6mg, 0.80 mmol) was added, and the reaction was completed by heating at 50°C. Adjust the pH to 7-8 with saturated sodium bicarbonate solution, leave it at room temperature overnight, concentrate, adjust the pH to about 11 with triethylamine, and then purify with a large plate (MeOH/DCM = 1/5) to obtain 153g of the compound 1c , yield 45.7%. ¹ H NMR (400 MHz, CDCl ₃ ): δ 8.90 (s, 2H), 8.57 (s, 1H), 3.72-3.45 (m, 4H), 3.11-2.40 (m, 12H), 2.17-1.85 (m, 4H), 1.71-1.46 (m, 8H), 1.37-0.96 (m, 72H), 0.86 (t, 12H).

步驟 3 ：化合物 1的製備

Step 3 : Preparation of compound 1

氮氣保護下，將化合物 1c(200 mg, 0.201 mmol)溶解於THF (5 mL)中，加入1滴DMF，加入SOCl ₂(598 mg, 5.02 mmol)，於室溫34℃下加熱反應3小時，減壓蒸出溶劑，油泵抽乾；粗品待用，氮氣保護下，在另外一個瓶中稱取化合物 E（177 mg, 0.26 mmol），用無水二氯甲烷（3 ml）溶解，加入三乙胺（202 mg, 2 mmol），將反應置於冰水浴中攪拌10~30分鐘，將前面得到的粗品一次性加入到該反應液中，自然升至室溫反應過夜，反應液直接藉由矽膠管柱（300-400目，二氯甲烷/甲醇 = 20:1-10:1）分離純化，得到83mg油狀化合物 1，收率26 %。 MS m/z (ESI):795.3[M/2+1] ⁺。 ¹H NMR (400 MHz,CDCl ₃): δ8.70 (s, 3 H), 8.52 (bs, 3 H), 4.92-4.83 (m, 2 H), 3.62-3.52 (m, 6 H), 3.12-2.82 (m, 18 H), 2.30-2.23 (m, 4 H), 2.18-2.02 (m, 6 H), 1.77-1.43 (m, 24 H), 1.39-1.18 (m, 100 H), 0.93-0.85(m, 24 H)。 Under nitrogen protection, compound 1c (200 mg, 0.201 mmol) was dissolved in THF (5 mL), 1 drop of DMF was added, SOCl ₂ (598 mg, 5.02 mmol) was added, and the reaction was heated at room temperature at 34°C for 3 hours. The solvent was evaporated under reduced pressure, and the oil pump was drained; the crude product was set aside, under the protection of nitrogen, weighed compound E (177 mg, 0.26 mmol) in another bottle, dissolved in anhydrous dichloromethane (3 ml), added triethylamine (202 mg, 2 mmol), the reaction was placed in an ice-water bath and stirred for 10-30 minutes, the crude product obtained above was added to the reaction solution at one time, and the reaction solution was naturally raised to room temperature overnight, and the reaction solution was directly passed through the silicone tube Column (300-400 mesh, dichloromethane/methanol = 20:1-10:1) was separated and purified to obtain 83 mg of oily compound 1 with a yield of 26%. MS m/z (ESI): 795.3 [M/2+1] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ): δ 8.70 (s, 3 H), 8.52 (bs, 3 H), 4.92-4.83 (m, 2 H), 3.62-3.52 (m, 6 H), 3.12- 2.82 (m, 18H), 2.30-2.23 (m, 4H), 2.18-2.02 (m, 6H), 1.77-1.43 (m, 24H), 1.39-1.18 (m, 100H), 0.93- 0.85 (m, 24 H).

實施例2 辛-4-基9-((3-(3,5-雙((3-(二(十二烷基胺基))丙基)胺基甲醯基)苯甲醯胺基)丙基)(十二烷基)胺基)壬酸酯

Example 2 Oct-4-yl 9-((3-(3,5-bis((3-(di(dodecylamino))propyl)aminoformyl)benzamido) Propyl)(dodecyl)amino)nonanoate

步驟 1 ：化合物 2b的製備

Step 1 : Preparation of compound 2b

於氮氣保護下，將化合物 E3(2.5 g, 7.15 mmol)和 2a(12.4 mg, 71.5 mmol)溶解於EtOH (10 mL)中，室溫（25-30℃）下反應24 h，反應液用石油醚和水分層，再用水洗2次，硫酸鈉乾燥，過濾濃縮，快速管柱層析層析，沖提劑MeOH/DCM，MeOH：0%→25%，得1.7 g油狀化合物 2b，產率53.8%。 MS m/z (ESI): 443.4[M+1] ⁺。 ¹H NMR (400 MHz,CDCl ₃)： δ5.20 (bs, 1 H), 4.92-4.83 (m, 1 H), 3.27-3.12 (m, 2 H), 2.70(t, 2 H), 2.60(t, 2 H), 2.27(t, 2 H), 2.20(bs, 1 H), 1.75-1.52 (m, 4 H), 1.51-1.42 (m, 15 H), 1.41-1.22 (m, 14 H), 0.93-0.85(m, 6 H)。 Under the protection of nitrogen, compound E3 (2.5 g, 7.15 mmol) and 2a (12.4 mg, 71.5 mmol) were dissolved in EtOH (10 mL), and reacted at room temperature (25-30 °C) for 24 h, and the reaction solution was purified with petroleum Ether and water were separated, washed twice with water, dried over sodium sulfate, concentrated by filtration, flash column chromatography, eluting agent MeOH/DCM, MeOH: 0%→25%, and 1.7 g of oily compound 2b was obtained. The rate is 53.8%. MS m/z (ESI): 443.4[M+1] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ): δ 5.20 (bs, 1 H), 4.92-4.83 (m, 1 H), 3.27-3.12 (m, 2 H), 2.70(t, 2 H), 2.60( t, 2H), 2.27(t, 2H), 2.20(bs, 1H), 1.75-1.52 (m, 4H), 1.51-1.42 (m, 15H), 1.41-1.22 (m, 14H ), 0.93-0.85(m, 6H).

步驟 2 ：化合物 2c的製備

Step 2 : Preparation of compound 2c

於氮氣保護下，將化合物 V1(675 mg, 2.71 mmol)和 2b(1.0 g, 2.26 mmol)溶解於DMF (4 mL)中，加入碳酸鉀(374 mg, 2.71 mmol)，80 ℃加熱反應過夜，反應液用乙酸乙酯稀釋，用水洗後，再用少量乙酸乙酯反萃水相，合併有機相，水洗3次，食鹽水洗一次，硫酸鈉乾燥，過濾濃縮，快速管柱層析純化，沖提體系MeOH/DCM，MeOH%：0%→25% (30 min)得0.92克油狀化合物 2c，產率66.7%。 ¹H NMR (400 MHz,CDCl ₃)： δ5.72 (bs, 1 H), 4.92-4.83 (m, 1 H), 3.27-3.12 (m, 2 H), 2.60-2.30(m, 4 H), 2.27(t, 2 H), 1.72-1.56 (m, 4 H), 1.51-1.22 (m, 51 H), 0.93-0.85(m, 9 H)。 Under nitrogen protection, compound V1 (675 mg, 2.71 mmol) and 2b (1.0 g, 2.26 mmol) were dissolved in DMF (4 mL), potassium carbonate (374 mg, 2.71 mmol) was added, and the reaction was heated at 80°C overnight. The reaction solution was diluted with ethyl acetate, washed with water, and the aqueous phase was back-extracted with a small amount of ethyl acetate, the organic phases were combined, washed three times with water, washed once with saline, dried over sodium sulfate, concentrated by filtration, purified by flash column chromatography, rinsed Extraction system MeOH/DCM, MeOH%: 0%→25% (30 min) to obtain 0.92 g of oily compound 2c , yield 66.7%. ¹ H NMR (400 MHz, CDCl ₃ ): δ 5.72 (bs, 1 H), 4.92-4.83 (m, 1 H), 3.27-3.12 (m, 2 H), 2.60-2.30 (m, 4 H), 2.27(t, 2H), 1.72-1.56(m, 4H), 1.51-1.22(m, 51H), 0.93-0.85(m, 9H).

步驟 3 ：化合物 2d的製備

Step 3 : Preparation of compound 2d

將化合物 2c(0.92 g, 1.5 mmol)溶解於1,4-二噁烷 (2 mL)中，HCl 4M in 1,4-二噁烷 (2 mL, 8 mmol)於室溫下加入，室溫（25-30℃）下反應，LCMS監測得反應基本完全，反應液直接濃縮並於油泵上抽2小時，所得粗品化合物 2d直接用於下一步。 MS m/z (ESI): 511.5[M+1] ⁺。 Compound 2c (0.92 g, 1.5 mmol) was dissolved in 1,4-dioxane (2 mL), HCl 4M in 1,4-dioxane (2 mL, 8 mmol) was added at room temperature, room temperature The reaction was carried out at (25-30°C), and the reaction was almost complete as monitored by LCMS. The reaction liquid was directly concentrated and pumped on an oil pump for 2 hours, and the obtained crude compound 2d was directly used in the next step. MS m/z (ESI): 511.5 [M+1] ⁺ .

步驟 4 ：化合物 2的製備

Step 4 : Preparation of compound 2

將化合物 1c(140 mg, 0.14 mmol)和 2d(107 mg, 0.18 mmol)溶解於DCM (2 mL)中，加入HATU (80 mg, 0.21 mmol)和三乙胺(39 μL, 0.28 mmol)，反應於室溫下攪拌，TLC監測反應基本完全，乙酸乙酯稀釋，用碳酸氫鈉溶液洗一次，水洗兩次，濃縮，製備板純化，展開劑(MeOH/DCM = 1/5)/(EA/PE = 1/1)=1.2/0.8，得113 mg油狀化合物 2，產率54.1%。純度：99.4%。 MS m/z (ESI): 1488.9[M+1] ⁺。 ¹H NMR (400 MHz, CDCl ₃): δ 8.79-8.21 (m, 6H), 4.97-4.82 (m, 1H), 3.68-3.50 (m, 6H), 2.91-2.41(m, 18H), 2.26 (t, J = 7.4 Hz, 2H), 1.94-1.71 (m, 6H), 1.66-1.12 (m, 122H), 0.96-0.81 (m, 21H)。 Compound 1c (140 mg, 0.14 mmol) and 2d (107 mg, 0.18 mmol) were dissolved in DCM (2 mL), HATU (80 mg, 0.21 mmol) and triethylamine (39 μL, 0.28 mmol) were added, and the reaction Stir at room temperature, TLC monitors that the reaction is almost complete, dilute with ethyl acetate, wash once with sodium bicarbonate solution, wash twice with water, concentrate, prepare a plate for purification, developing solvent (MeOH/DCM = 1/5)/(EA/ PE = 1/1)=1.2/0.8, 113 mg of oily compound 2 was obtained, and the yield was 54.1%. Purity: 99.4%. MS m/z (ESI): 1488.9 [M+1] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ): δ 8.79-8.21 (m, 6H), 4.97-4.82 (m, 1H), 3.68-3.50 (m, 6H), 2.91-2.41(m, 18H), 2.26 ( t, J = 7.4 Hz, 2H), 1.94-1.71 (m, 6H), 1.66-1.12 (m, 122H), 0.96-0.81 (m, 21H).

實施例3 2,2'-((3-(3,5-雙((3-(二(十二烷基胺基))丙基)胺基甲醯基)苯甲醯胺基)丙基)氮烷基二基)二乙酸二壬酯

Example 3 2,2'-((3-(3,5-bis((3-(di(dodecylamino))propyl)aminoformyl)benzamido)propyl ) azaalkyldiyl) dinonyl diacetate

步驟 1：化合物 3b的製備

Step 1 : Preparation of compound 3b

將化合物 3a(1.0 g, 4.25 mmol，1.0 eq)和 V3（0.32 g，1.85 mmol，2.3 eq）溶於ACN(10 mL)，然後依次加入碳酸鉀（0.59 g, 1.85 mmol, 2.3 eq）和KI(0.71 g, 1.85 mmol, 2.3 eq，氮氣保護下在75℃下攪拌4-10 h，反應液減壓濃縮除去溶劑，殘留物加入EA（30 ml）溶解後用飽和碳酸氫鈉洗（20 ml×2）、飽和食鹽水（20 mL）洗一次，用無水硫酸鈉乾燥，減壓濃縮，用管柱層析純化（100-200目，PE/EA=5:1）得到750 mg化合物 3b，收率37%。 MS m/z (ESI): 543.5[M+1] ⁺。 ¹H NMR (400 MHz, CDCl ₃): δ 4.11-4.08 (m, 4 H), 3.50 (s, 4 H), 2.75 (s, 2 H), 1.64-1.61 (m, 8 H), 1.43 (s, 9 H), 1.30-1.26 (m, 25 H), 0.89-0.85 (m, 6 H)。 Compound 3a (1.0 g, 4.25 mmol, 1.0 eq) and V3 (0.32 g, 1.85 mmol, 2.3 eq) were dissolved in ACN (10 mL), then potassium carbonate (0.59 g, 1.85 mmol, 2.3 eq) and KI were added sequentially (0.71 g, 1.85 mmol, 2.3 eq, stirred at 75°C for 4-10 h under nitrogen protection, the reaction solution was concentrated under reduced pressure to remove the solvent, the residue was dissolved in EA (30 ml) and washed with saturated sodium bicarbonate (20 ml ×2), washed once with saturated saline (20 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, purified by column chromatography (100-200 mesh, PE/EA=5:1) to obtain 750 mg of compound 3b , Yield 37%. MS m/z (ESI): 543.5[M+1] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ): δ 4.11-4.08 (m, 4 H), 3.50 (s, 4 H) , 2.75 (s, 2 H), 1.64-1.61 (m, 8 H), 1.43 (s, 9 H), 1.30-1.26 (m, 25 H), 0.89-0.85 (m, 6 H).

步驟 2：化合物 3c的製備

Step 2 : Preparation of Compound 3c

化合物 3b（0.75 g，1.38 mmol, 1.0 eq）溶解於1,4-二噁烷 (4 mL)中，於室溫下加入HCl 4M的1,4-二噁烷溶液 (16 ml)，攪拌反應，LCMS檢測顯示反應完成。直接減壓濃縮至乾得到659 mg化合物 3c，收率：100%。 MS m/z (ESI): 443.4[M+1] ⁺。 Compound 3b (0.75 g, 1.38 mmol, 1.0 eq) was dissolved in 1,4-dioxane (4 mL), and HCl 4M in 1,4-dioxane solution (16 ml) was added at room temperature, and the reaction was stirred , LCMS detection showed the reaction was complete. Directly concentrated to dryness under reduced pressure to obtain 659 mg of compound 3c , yield: 100%. MS m/z (ESI): 443.4[M+1] ⁺ .

步驟 3：化合物 3的製備

Step 3 : Preparation of compound 3

將化合物 3c（0.66 g，1.24 mmol, 1.3 eq）和化合物 1c（1.02 g, 0.96 mmol， 1.0 eq）溶於DCM（20 ml）中，然後依次加入HOBT（0.16 g, 1.15 mmol, 1.2 eq）、EDCI(0.37 g, 1.91 mmol, 2.0 eq)和DIEPA（0.62 g， 4.78 mmol， 5.0 eq），氮氣保護下攪拌15~20h，反應液減壓濃縮除去溶劑，殘留物加入EA（30 ml）溶解後用飽和碳酸氫鈉洗（20 ml×2）、飽和食鹽水（20 mL）洗一次，用無水硫酸鈉乾燥，減壓濃縮，用反相管柱層析純化（H ₂O/ACN，95%出產物）得到1.02 g化合物 3，收率77.78%，純度：96.2%。 MS m/z (ESI): 1420.9[M+1] ⁺。 ¹H NMR (400 MHz, CD3OD): δ 8.49 (s, 3 H), 4.12-4.06 (m, 4 H), 3.63-3.48 (m, 9 H), 3.13-2.81 (m, 12 H), 2.01-1.97 (m, 6 H), 1.80-1.78 (m, 2 H), 1.63-1.58 (m, 12 H), 1.34-1.22(m,98H), 0.91-0.87 (m,18H)。 Compound 3c (0.66 g, 1.24 mmol, 1.3 eq) and compound 1c (1.02 g, 0.96 mmol, 1.0 eq) were dissolved in DCM (20 ml), and then HOBT (0.16 g, 1.15 mmol, 1.2 eq) and EDCI (0.37 g, 1.91 mmol, 2.0 eq) and DIEPA (0.62 g, 4.78 mmol, 5.0 eq), stirred under nitrogen for 15~20h, the reaction solution was concentrated under reduced pressure to remove the solvent, and the residue was dissolved in EA (30 ml) Wash with saturated sodium bicarbonate (20 ml×2) and saturated brine (20 mL) once, dry over anhydrous sodium sulfate, concentrate under reduced pressure, and purify by reverse-phase column chromatography (H ₂ O/ACN, 95% product) to obtain 1.02 g of compound 3 with a yield of 77.78% and a purity of 96.2%. MS m/z (ESI): 1420.9 [M+1] ⁺ . ¹ H NMR (400 MHz, CD3OD): δ 8.49 (s, 3 H), 4.12-4.06 (m, 4 H), 3.63-3.48 (m, 9 H), 3.13-2.81 (m, 12 H), 2.01 -1.97 (m, 6H), 1.80-1.78 (m, 2H), 1.63-1.58 (m, 12H), 1.34-1.22(m,98H), 0.91-0.87 (m,18H).

實施例4 ((((苯-1,3,5-三羰基)三(氮烷基二基))三(丁烷-4,1-二基))三(氮烷基三基))六(己烷-6,1-二基)六(2-己基癸酸酯) )

Example 4 ((((benzene-1,3,5-tricarbonyl) three (azanyl diyl)) three (butane-4,1-diyl)) three (azanyl triyl)) six (Hexane-6,1-diyl)hexa(2-hexyldecanoate) )

步驟 1 ：化合物 4c的製備 Step 1 : Preparation of compound 4c

於氮氣保護下，將化合物 4a(6.4 g, 35.3 mmol)和 4b(10.0 g, 39.0 mmol)溶解於DCM (64 mL)中，加入DMAP (1.3 g, 10.6 mmol)和EDCI (10.2 g, 53.2 mmol)，室溫下反應，TLC（EA/PE = 1/10）顯示反應完全，濃縮，純化得8.9 g油狀化合物 4c，產率：57.8%。 ¹H NMR (400 MHz, CDCl ₃): δ 4.07 (t, J= 6.6 Hz, 2H), 3.40 (t, J= 6.8 Hz, 2H), 2.35-2.26 (m, 1H), 1.93-1.81 (m, 2H), 1.70-1.15 (m, 30H), 0.87 (t, J= 6.3 Hz, 6H)。 Under nitrogen protection, compounds 4a (6.4 g, 35.3 mmol) and 4b (10.0 g, 39.0 mmol) were dissolved in DCM (64 mL), DMAP (1.3 g, 10.6 mmol) and EDCI (10.2 g, 53.2 mmol) were added ), reacted at room temperature, TLC (EA/PE = 1/10) showed that the reaction was complete, concentrated and purified to obtain 8.9 g of oily compound 4c , yield: 57.8%. ¹ H NMR (400 MHz, CDCl ₃ ): δ 4.07 (t, J = 6.6 Hz, 2H), 3.40 (t, J = 6.8 Hz, 2H), 2.35-2.26 (m, 1H), 1.93-1.81 (m , 2H), 1.70-1.15 (m, 30H), 0.87 (t, J = 6.3 Hz, 6H).

步驟 2：化合物 4e的製備

Step 2 : Preparation of Compound 4e

於氮氣保護下，將化合物 4c(5.9 g, 14.1 mmol)和 4d(1.0 g, 5.7 mmol)溶解於DMF (10 mL)中，加入K ₂CO ₃(1.9 g, 13.7 mmol)，80℃反應，TLC（MeOH/DCM = 1/10）檢測基本反應完全，乙酸乙酯（100 mL）稀釋，用水洗，有機相硫酸鈉乾燥，過濾濃縮，純化得0.96 g油狀化合物 4e，產率19.3%。 ¹H NMR (400 MHz, CDCl ₃): δ 5.00 (s, 1H), 4.06 (t, J= 6.7 Hz, 4H), 3.18-3.04 (m, 2H), 2.06-2.16 (m, 8H), 1.72-1.52 (m, 12H), 1.52-1.10 (m, 65H), 0.87 (t, J= 6.1 Hz, 12H)。 Under the protection of nitrogen, compounds 4c (5.9 g, 14.1 mmol) and 4d (1.0 g, 5.7 mmol) were dissolved in DMF (10 mL), and K ₂ CO ₃ (1.9 g, 13.7 mmol) was added to react at 80°C. TLC (MeOH/DCM = 1/10) detected that the basic reaction was complete, diluted with ethyl acetate (100 mL), washed with water, dried the organic phase over sodium sulfate, filtered and concentrated, and purified to obtain 0.96 g of oily compound 4e with a yield of 19.3%. ¹ H NMR (400 MHz, CDCl ₃ ): δ 5.00 (s, 1H), 4.06 (t, J = 6.7 Hz, 4H), 3.18-3.04 (m, 2H), 2.06-2.16 (m, 8H), 1.72 -1.52 (m, 12H), 1.52-1.10 (m, 65H), 0.87 (t, J = 6.1 Hz, 12H).

步驟 3 ：化合物 4f的製備

Step 3 : Preparation of Compound 4f

室溫下，將化合物 4e(0.96 g, 1.1 mmol)溶解於1,4-二噁烷(5 mL)中，加入HCl(4M 1,4-二噁烷，5 mL, 20.0 mmol)，室溫下反應2~6小時，反應液直接濃縮至乾，所得粗品直接用於下一步。 At room temperature, the compound 4e (0.96 g, 1.1 mmol) was dissolved in 1,4-dioxane (5 mL), added HCl (4M 1,4-dioxane, 5 mL, 20.0 mmol), room temperature The reaction was carried out for 2-6 hours, the reaction solution was directly concentrated to dryness, and the obtained crude product was directly used in the next step.

步驟 4 ：化合物 4的製備

Step 4 : Preparation of Compound 4

於氮氣保護下，將粗品化合物 4f(約1.1 mmol)溶解於DCM (10 mL)中，加入化合物 4g(84 mg, 0.32 mmol)，加入三乙胺(354 μL, 2.55 mmol)，於室溫下反應15~18小時，將反應液與矽膠（100-200目）拌樣濃縮，200-300目矽膠管柱層析(MeOH/DCM = 1/10)/(EA/PE = 1/1) = 3/1得到粗品670 mg，取約320 mg部分進一步大板純化，展開劑為(MeOH/DCM = 1/5)/(EA/PE = 1/1) = 3/1，得202 mg油狀化合物 4，兩步收率26.0%。 ¹H NMR (400 MHz, CDCl ₃): δ 8.40 (s, 3H), 7.04 (s, 3H), 4.04 (t, J= 6.6 Hz, 12H), 3.55-3.41 (m, 6H), 2.62-2.36 (m, 18H), 2.36-2.11 (m, 12H), 1.74-1.50 (m, 36 H), 1.50-1.09 (m, 162H), 0.87 (t, J= 6.0 Hz, 36H)。 Under nitrogen protection, the crude compound 4f (about 1.1 mmol) was dissolved in DCM (10 mL), compound 4g (84 mg, 0.32 mmol) was added, triethylamine (354 μL, 2.55 mmol) was added, and at room temperature React for 15-18 hours, mix the reaction liquid with silica gel (100-200 mesh) and concentrate, perform column chromatography on 200-300 mesh silica gel (MeOH/DCM = 1/10)/(EA/PE = 1/1) = 3/1 obtained 670 mg of crude product, about 320 mg was taken for further large-scale purification, the developer was (MeOH/DCM = 1/5)/(EA/PE = 1/1) = 3/1, and 202 mg of oil was obtained Compound 4 , the two-step yield is 26.0%. ¹ H NMR (400 MHz, CDCl ₃ ): δ 8.40 (s, 3H), 7.04 (s, 3H), 4.04 (t, J = 6.6 Hz, 12H), 3.55-3.41 (m, 6H), 2.62-2.36 (m, 18H), 2.36-2.11 (m, 12H), 1.74-1.50 (m, 36H), 1.50-1.09 (m, 162H), 0.87 (t, J = 6.0 Hz, 36H).

實施例5 N-(3-(3,5-雙((3-(二(十二烷基胺基))丙基)胺基甲醯基)苯甲醯胺基)丙基)-N-十二烷基甘胺酸壬基酯

Example 5 N-(3-(3,5-bis((3-(di(dodecylamino))propyl)aminoformyl)benzamido)propyl)-N- Nonyl Lauryl Glycinate

步驟 1 ：化合物 5c的製備

Step 1 : Preparation of compound 5c

在25 mL三口瓶中，依次加入化合物 5a(1.3 g, 3.8 mmol)，DMF (13 mL)，化合物 5b(1.0 g, 3.8 mmol)和碳酸鉀(524 mg, 3.8 mmol)，氮氣保護下置換三次，開啟攪拌，加熱內溫110℃ 2h。取樣LC-MS檢測顯示有目標產物的質譜峰，原料少量剩餘。停止反應，將反應液降溫到25℃，加入150 mL的水，隨後用EtOAc（60 mL × 3）萃取，有機相合併，依次水洗（60 mL），飽和食鹽水洗（60 mL），無水硫酸鈉（5 g）乾燥，過濾蒸乾得到粗品。經過矽膠管柱層析（PE/EA=20/1-10/1-5/1）分離得到油狀化合物 5c（1.860 g，收率：93.0%）。 Ms (ESI)：m/z 527.5 [M+H] ⁺。 In a 25 mL three-necked flask, compound 5a (1.3 g, 3.8 mmol), DMF (13 mL), compound 5b (1.0 g, 3.8 mmol) and potassium carbonate (524 mg, 3.8 mmol) were sequentially added, and replaced three times under nitrogen protection , start stirring, and heat the inner temperature to 110°C for 2h. Sampling LC-MS detection showed the mass spectrum peak of the target product, and a small amount of raw materials remained. Stop the reaction, cool the reaction solution to 25°C, add 150 mL of water, then extract with EtOAc (60 mL × 3), combine the organic phases, wash with water (60 mL), saturated brine (60 mL), anhydrous sodium sulfate (5 g) was dried, filtered and evaporated to obtain the crude product. The oily compound 5c (1.860 g, yield: 93.0%) was separated by silica gel column chromatography (PE/EA=20/1-10/1-5/1). Ms (ESI): m/z 527.5 [M+H] ⁺ .

步驟 2 ：化合物 5d的製備

Step 2 : Preparation of compound 5d

在25 mL單口瓶中，依次加入化合物 5c(1.7 g, 3.2 mmol)，4 N HCl(二噁烷中) (10 mL)，氮氣保護下置換一次，開啟攪拌，室溫23-26℃攪拌2h，取樣LC-MS檢測顯示原料反應完畢。停止反應，減壓濃縮得到白色固體化合物 5d（1.8 g,收率：111.6%）。 Ms (ESI)：m/z 427.4 [M+H] ⁺。 In a 25 mL single-necked bottle, add compound 5c (1.7 g, 3.2 mmol), 4 N HCl (in dioxane) (10 mL) in sequence, replace once under nitrogen protection, start stirring, and stir at room temperature 23-26 ° C for 2 h , Sampling LC-MS detection showed that the reaction of raw materials was complete. The reaction was stopped and concentrated under reduced pressure to obtain white solid compound 5d (1.8 g, yield: 111.6%). Ms (ESI): m/z 427.4 [M+H] ⁺ .

步驟 3 ：化合物 5的製備

Step 3 : Preparation of Compound 5

在100 mL三口瓶中，依次加入化合物 1c(2.0 g, 2.01 mmol)，無水DCM (40 mL)，化合物 5d(1.2 g, 2.41 mmol)和DIEA (2.1 mL, 12 mmol)，氬氣保護下置換三次，開啟攪拌，冰水浴冷卻到5℃。一次性加入HATU (917 mg, 2.41 mmol)內溫小於10 ℃攪拌30分鐘，然後室溫25-32 ℃攪拌19h，取樣HPLC檢測顯示原料反應完畢，停止反應，加入50 mL二氯甲烷稀釋，一次用飽和碳酸氫鈉溶液（30 mL）和飽和食鹽水洗，無水硫酸鈉（5 g）乾燥，過濾蒸乾得到粗品。經過200-300目的正相矽膠管柱層析（PE/EtOH=0%-60%）得到油狀物，然後Pre-TLC（DCM/MeOH=20/1）得到油狀化合物 5（526 mg，收率：18.6%，HPLC純度94.63%）。 MS m/z (ESI): 1404.8[M+1] ⁺。 ¹H NMR (400 MHz,CDCl ₃): δ 8.47 (s, 3 H), 8.33 (s, 3 H), 4.10-4.02 (t, 2 H), 3.66-3.52 (m, 6 H), 3.32 (s, 2 H), 2.70-2.48 (m, 16 H), 1.74-1.43 (m, 18 H), 1.33-1.29 (m, 12 H), 1.27-1.14 (m, 90 H), 0.89-0.83(m, 18 H)。 In a 100 mL three-neck flask, sequentially add compound 1c (2.0 g, 2.01 mmol), anhydrous DCM (40 mL), compound 5d (1.2 g, 2.41 mmol) and DIEA (2.1 mL, 12 mmol), and replace under argon Three times, start stirring, and cool to 5°C in an ice-water bath. Add HATU (917 mg, 2.41 mmol) at one time and stir for 30 minutes at an internal temperature of less than 10 °C, then stir at room temperature 25-32 °C for 19 h, sample HPLC detection shows that the reaction of raw materials is complete, stop the reaction, add 50 mL of dichloromethane to dilute, once Wash with saturated sodium bicarbonate solution (30 mL) and saturated brine, dry over anhydrous sodium sulfate (5 g), filter and evaporate to dryness to obtain the crude product. After 200-300 mesh normal-phase silica gel column chromatography (PE/EtOH=0%-60%), the oily substance was obtained, and then Pre-TLC (DCM/MeOH=20/1) gave the oily compound 5 (526 mg, Yield: 18.6%, HPLC purity 94.63%). MS m/z (ESI): 1404.8 [M+1] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ): δ 8.47 (s, 3 H), 8.33 (s, 3 H), 4.10-4.02 (t, 2 H), 3.66-3.52 (m, 6 H), 3.32 ( s, 2H), 2.70-2.48 (m, 16H), 1.74-1.43 (m, 18H), 1.33-1.29 (m, 12H), 1.27-1.14 (m, 90H), 0.89-0.83( m, 18H).

對比實施例1

Comparative Example 1

對比化合物1 採用“ Nano Lett.2015, 15, 8099-8107和WO2016187531A1”中方法製備而得。 Comparative compound 1 was prepared by the method in " Nano Lett. 2015, 15, 8099-8107 and WO2016187531A1".

對比實施例2

Comparative Example 2

對比化合物2 採用“WO2019099501A1”中方法製備而得。 Comparative compound 2 It is prepared by the method in "WO2019099501A1".

測試例1：脂質顆粒組成物遞送能力Test Example 1: Delivery Ability of Lipid Particle Composition

1.1 製備方法分別將化合物1、2、3、4和對比化合物1、2溶於乙醇溶液，與溶於乙醇的DOPE、膽固醇、DMG-PEG溶液，以20：30：40：0.75的莫耳比混合配製乙醇脂質溶液。將編碼表達螢光素酶的mRNA （GenBank: MN728548.1）溶於檸檬酸鹽緩衝液，配製mRNA水溶液。將乙醇脂質溶液和mRNA水溶液藉由微流體混合，總脂質與mRNA的重量比約為12-36：1製備脂質體。在PBS溶液中透析去除乙醇，得到包封編碼螢光素酶mRNA的脂質體納米顆粒（LNP）製劑。 1.1 Preparation method Compounds 1, 2, 3, 4 and comparative compounds 1, 2 were dissolved in ethanol solution, and mixed with DOPE, cholesterol, DMG-PEG solution dissolved in ethanol at a molar ratio of 20:30:40:0.75 to prepare ethanol lipid solution. Dissolve mRNA encoding luciferase (GenBank: MN728548.1) in citrate buffer to prepare mRNA aqueous solution. The ethanol lipid solution and the mRNA aqueous solution were mixed by microfluidics, and the weight ratio of total lipid to mRNA was about 12-36:1 to prepare liposomes. Ethanol was dialyzed in PBS solution to obtain a liposome nanoparticle (LNP) preparation encapsulating luciferase mRNA.

1.2 脂質奈米顆粒組成物的表徵使用Malvern Zetasizer Nano ZS，以173°反向散射檢測模式，利用動態光散射檢測脂質體奈米顆粒的奈米尺寸和多分散係數（PDI）。 1.2 Characterization of lipid nanoparticles composition The nanometer size and polydispersity index (PDI) of liposomal nanoparticles were detected by dynamic light scattering using a Malvern Zetasizer Nano ZS in 173° backscatter detection mode.

使用Quant-iT RiboGreen RNA Assay Kit RNA定量檢測試劑盒（購於賽默飛世爾科技，貨號R11490），測定脂質體包封率。Quant-iT RiboGreen RNA Assay Kit RNA Quantitative Detection Kit (purchased from Thermo Fisher Scientific, catalog number R11490) was used to determine the liposome encapsulation efficiency.

使用基於6-(p-甲苯胺)-2-萘磺酸鈉鹽（TNS）的螢光分析，測定脂質體奈米顆粒中陽離子的pKa。配製150 mM NaCl，10 mM 磷酸鈉，10 mM 檸檬酸鈉，10 mM硼酸鈉，pH 3-11.5不同pH的緩衝液。配製300 μM TNS溶液，加入到緩衝液中。將脂質奈米顆粒分別加入到不同pH的緩衝液中，充分混合後，使用螢光酶標儀在室溫下檢測激發波長325 nm，發射波長435 nm的螢光強度。對螢光數據擬合分析，pKa為產生半數最大螢光強度的pH值。相關數據見表1。The pKa of cations in liposomal nanoparticles was determined using a fluorometric assay based on 6-(p-toluidine)-2-naphthalenesulfonic acid sodium salt (TNS). Prepare 150 mM NaCl, 10 mM sodium phosphate, 10 mM sodium citrate, 10 mM sodium borate, pH 3-11.5 buffers with different pH. Prepare a 300 μM TNS solution and add it to the buffer. Lipid nanoparticles were added to buffer solutions with different pHs, and after mixing thoroughly, the fluorescence intensity at an excitation wavelength of 325 nm and an emission wavelength of 435 nm was detected at room temperature using a fluorescent microplate reader. For fluorescence data fitting analysis, pKa is the pH value that produces half the maximum fluorescence intensity. See Table 1 for relevant data.

表1 編號尺寸（nm） PDI 包封率（%） pKa 化合物1 128.7 0.08 99.8 6.28 化合物2 151 0.05 99.8 6.16 化合物3 116 0.13 99.0 6.15 化合物4 185.2 0.05 98.1 6.03 對比化合物1 145.5 0.11 98.3 6.46 對比化合物2 149.1 0.08 99.1 6.13 Table 1 serial number Size (nm) PDI Encapsulation rate (%) pKa Compound 1 128.7 0.08 99.8 6.28 Compound 2 151 0.05 99.8 6.16 Compound 3 116 0.13 99.0 6.15 Compound 4 185.2 0.05 98.1 6.03 Comparative compound 1 145.5 0.11 98.3 6.46 Comparative compound 2 149.1 0.08 99.1 6.13

1.3評估脂質顆粒組成物在細胞水平的mRNA表達效率以對比化合物1和2對應的脂質體奈米顆粒為對照，檢測化合物1-3對應的脂質體奈米顆粒的體外細胞水平的mRNA表達效率。 1.3 Evaluate the mRNA expression efficiency of lipid particle composition at the cellular level The liposome nanoparticles corresponding to compounds 1 and 2 were used as controls to detect the mRNA expression efficiency of liposome nanoparticles corresponding to compounds 1-3 at the cellular level in vitro.

將HEK 293細胞接種到細胞孔板培養過夜，待細胞密度達到80%以上，將包封螢光素酶mRNA的脂質體LNP溶液加入到細胞板孔培養基中。24小時後，使用螢光素酶報告基因檢測試劑盒（Promega）和酶標儀檢測表達螢光素酶蛋白的螢光強度。螢光強度值即酶標儀檢測到的螢光數值。每個化合物對應的LNP至少3組重複計算平均螢光值強度和統計學差異，數據見表2和圖1。HEK 293 cells were inoculated into the cell well plate and cultured overnight, and when the cell density reached above 80%, the liposome LNP solution encapsulating luciferase mRNA was added to the culture medium of the cell plate well. After 24 hours, the fluorescence intensity of the expressed luciferase protein was detected using a luciferase reporter gene detection kit (Promega) and a microplate reader. The fluorescence intensity value is the fluorescence value detected by the microplate reader. For each compound, the LNP corresponding to at least three groups repeatedly calculated the average fluorescence value intensity and statistical difference, and the data are shown in Table 2 and Figure 1.

圖1中*即統計學0.01＜P＜0.05，表示組間具有統計學顯著性差異，**即統計學P＜0.01，***即統計學P＜0.001表示極顯著性差異。In Fig. 1, * means statistically 0.01<P<0.05, indicating statistically significant difference between groups, ** means statistically P<0.01, *** means statistically P<0.001 means extremely significant difference.

表2 編號平均螢光強度值化合物1 6.47E+06 化合物2 4.09E+06 化合物3 4.58E+06 對比化合物1 3.38E+06 對比化合物2 2.61E+06 Table 2 serial number Mean Fluorescence Intensity Value Compound 1 6.47E+06 Compound 2 4.09E+06 Compound 3 4.58E+06 Comparative compound 1 3.38E+06 Comparative compound 2 2.61E+06

結論：如表2和圖1所示螢光強度，化合物1、2和3對應的脂質奈米顆粒在細胞中遞送mRNA表達螢光素酶表達量優於對比化合物1和對比化合物2。Conclusion: As the fluorescence intensity shown in Table 2 and Figure 1, the lipid nanoparticles corresponding to compounds 1, 2 and 3 deliver mRNA expression luciferase expression in cells is better than comparative compound 1 and comparative compound 2.

1.4 評估脂質顆粒組成物體內肌肉注射遞送mRNA表達效率為評估脂質體奈米顆粒在體內有效遞送mRNA並表達相應編碼的蛋白質，以0.25 mg/kg的劑量對6-8週齡雌性BALB/c大腿肌肉部位注射包封有表達螢光素酶的mRNA脂質體奈米顆粒。6小時後，分別向每隻小鼠腹腔注射螢光素酶受質，使用IVIS小動物光學活體成像儀器（PerkinElme），拍攝小鼠的螢光圖片，並統計注射部位的螢光強度。螢光強度的高低代表螢光素酶蛋白的表達量高低，即反應脂質體奈米顆粒體內遞送mRNA的效率。表3和圖2中螢光強度即IVIS小動物光學活體成像儀拍攝統計的小鼠注射部位的螢光強度。每個化合物對應的LNP至少3組重複計算平均螢光值強度。 1.4 Evaluation of mRNA expression efficiency of intramuscular injection of lipid particle composition In order to evaluate the effective delivery of liposome nanoparticles to mRNA and express the corresponding encoded protein in vivo, 6-8 weeks old female BALB/c female BALB/c was injected with luciferase-encapsulated mRNA at the thigh muscle site at a dose of 0.25 mg/kg Liposome nanoparticles. Six hours later, luciferase substrates were injected intraperitoneally into each mouse, and IVIS small animal optical in vivo imaging instrument (PerkinElme) was used to take fluorescent pictures of the mice, and the fluorescence intensity at the injection site was counted. The level of fluorescence intensity represents the level of expression of luciferase protein, which reflects the efficiency of delivering mRNA in vivo by liposome nanoparticles. The fluorescence intensity in Table 3 and Figure 2 is the fluorescence intensity at the injection site of the mouse captured and counted by the IVIS small animal optical in vivo imager. At least 3 groups of LNP corresponding to each compound were repeated to calculate the average fluorescence intensity.

以對比化合物1對應的脂質體奈米顆粒為對照，檢測化合物1、2和3對應的脂質體奈米顆粒的體內肌肉注射mRNA遞送效率。相關數據見表3和圖2。Taking the liposome nanoparticle corresponding to compound 1 as a control, the in vivo intramuscular injection mRNA delivery efficiency of liposome nanoparticle corresponding to compound 1, 2 and 3 was detected. See Table 3 and Figure 2 for relevant data.

圖2中*即統計學0.01＜P＜0.05，表示組間具有統計學顯著性差異，**即統計學P＜0.01，***即統計學P＜0.001表示極顯著性差異。In Fig. 2, * means statistically 0.01<P<0.05, indicating statistically significant difference between groups, ** means statistically P<0.01, *** means statistically P<0.001 means extremely significant difference.

表3 編號平均螢光強度值化合物1 1.80E+08 化合物2 1.55E+08 化合物3 4.65E+07 對比化合物1 4.19E+07 結論：如表3和圖2所示螢光強度，化合物1、2和3對應的脂質奈米顆粒體內肌肉注射遞送mRNA，效果優於對比化合物1。 table 3 serial number Mean Fluorescence Intensity Value Compound 1 1.80E+08 Compound 2 1.55E+08 Compound 3 4.65E+07 Comparative compound 1 4.19E+07 Conclusion: As the fluorescence intensity shown in Table 3 and Figure 2, the lipid nanoparticles corresponding to compounds 1, 2 and 3 are better than the comparison compound 1 in delivering mRNA in vivo by intramuscular injection.

1.5 評估脂質顆粒體內肌肉注射遞送mRNA後的蛋白表達量為評估脂質體奈米顆粒將mRNA在體內有效遞送mRNA並表達相應編碼的蛋白質，以0.05 mg/kg的劑量對6-8週齡雌性BALB/c大腿肌肉部位注射包封有表達生長因子的mRNA（NCBI Reference Sequence: NM_000601.6）脂質體奈米顆粒。24小時後取注射部位肌肉，經研磨裂解後，使用ELISA試劑盒檢測生長因子蛋白表達量（pg/mg），即單位肌肉組織總蛋白量對應的生長因子蛋白量。每個化合物對應的LNP至少3組重複計算平均蛋白濃度。 1.5 Evaluation of protein expression after intramuscular injection of lipid particles delivered mRNA In order to evaluate the effective delivery of mRNA in vivo by liposome nanoparticles and the expression of the corresponding encoded protein, 6-8 week old female BALB/c thigh muscle was injected with encapsulated mRNA expressing growth factors at a dose of 0.05 mg/kg (NCBI Reference Sequence: NM_000601.6) Liposome nanoparticles. After 24 hours, the muscle at the injection site was taken, ground and lysed, and the expression level of growth factor protein (pg/mg) was detected using an ELISA kit, that is, the amount of growth factor protein corresponding to the total protein amount of muscle tissue. The average protein concentration was calculated for at least 3 groups of LNP corresponding to each compound.

以對比化合物1對應的脂質體奈米顆粒為對照，檢測化合物1和3對應的脂質體奈米顆粒的體內肌肉注射mRNA後的蛋白表達量，即mRNA體內肌肉注射的遞送效率。相關數據見表4和圖3。Taking the liposome nanoparticle corresponding to compound 1 as a control, the protein expression level of the liposome nanoparticle corresponding to compound 1 and 3 after intramuscular injection of mRNA was detected, that is, the delivery efficiency of mRNA in vivo intramuscular injection. See Table 4 and Figure 3 for relevant data.

圖3中*即統計學0.01＜P＜0.05，表示組間具有統計學顯著性差異，**即統計學P＜0.01，***即統計學P＜0.001表示極顯著性差異。In Fig. 3, * means statistically 0.01<P<0.05, indicating statistically significant difference between groups, ** means statistically P<0.01, *** means statistically P<0.001 means extremely significant difference.

表4 編號平均蛋白表達量（pg/mg）化合物1 121.4 化合物3 94.7 對比化合物1 43.6 結論：如表4和圖3所示蛋白表達量，化合物1和3對應的LNP體內肌肉注射遞送mRNA，蛋白表達量顯著高於對比化合物1。 Table 4 serial number Average protein expression (pg/mg) Compound 1 121.4 Compound 3 94.7 Comparative compound 1 43.6 Conclusion: As shown in Table 4 and Figure 3, the protein expression levels of the LNPs corresponding to compounds 1 and 3 delivered by intramuscular injection in vivo were significantly higher than those of the comparison compound 1.

1.6 評估脂質顆粒組成物體內尾靜脈注射mRNA表達效率為評估脂質體奈米顆粒將mRNA在體內有效遞送mRNA並表達相應編碼的蛋白質，以0.5 mg/kg的劑量對6-8週齡雌性BALB/c尾靜脈注射包封有表達螢光素酶的mRNA脂質體奈米顆粒。6小時後，分別向每隻小鼠腹腔注射螢光素酶受質，使用IVIS小動物光學活體成像儀器（PerkinElme），拍攝小鼠的螢光圖片，並統計小鼠全身的螢光強度。螢光強度的高低代表螢光素酶蛋白的表達量高低，即反應脂質體奈米顆粒體內遞送mRNA的效率。每個化合物對應的LNP至少3組重複計算平均螢光值強度，數據見表5和圖4。 1.6 Evaluate the mRNA expression efficiency of lipid particle composition in vivo tail vein injection In order to evaluate the effective delivery of mRNA and the expression of the corresponding encoded protein by liposome nanoparticles in vivo, 6-8 week-old female BALB/c tail vein was injected with encapsulated luciferase-expressing protein at a dose of 0.5 mg/kg mRNA liposome nanoparticles. Six hours later, luciferase substrates were injected intraperitoneally into each mouse, and IVIS small animal optical in vivo imaging instrument (PerkinElme) was used to take fluorescent pictures of the mice, and the fluorescence intensity of the whole body of the mice was counted. The level of fluorescence intensity represents the level of expression of luciferase protein, which reflects the efficiency of delivering mRNA in vivo by liposome nanoparticles. For each compound, at least three groups of LNPs were used to repeatedly calculate the average fluorescence intensity, and the data are shown in Table 5 and Figure 4.

圖4中螢光強度即IVIS小動物光學活體成像儀拍攝統計的小鼠全身的螢光強度。The fluorescence intensity in Figure 4 is the fluorescence intensity of the whole body of the mouse captured and counted by the IVIS small animal optical in vivo imager.

以對比化合物1對應的脂質體奈米顆粒為對照，檢測化合物1和2對應的脂質體奈米顆粒的體內尾靜脈注射的mRNA遞送效率。Using the liposome nanoparticle corresponding to comparative compound 1 as a control, the mRNA delivery efficiency of the in vivo tail vein injection of the liposome nanoparticle corresponding to compound 1 and 2 was detected.

圖4中*即統計學0.01＜P＜0.05，表示組間具有統計學顯著性差異，**即統計學P＜0.01，***即統計學P＜0.001表示極顯著性差異。In Fig. 4, * means statistically 0.01<P<0.05, indicating statistically significant difference between groups, ** means statistically P<0.01, *** means statistically P<0.001 means extremely significant difference.

表5 編號平均螢光強度值化合物1 7.47E+08 化合物2 9.69E+08 對比化合物1 3.98E+08 結論：如表5和圖4所示螢光強度，化合物1和2對應的mRNA脂質奈米顆粒尾靜脈注射，蛋白表達量顯著優於對比化合物1。 table 5 serial number Mean Fluorescence Intensity Value Compound 1 7.47E+08 Compound 2 9.69E+08 Comparative compound 1 3.98E+08 Conclusion: As the fluorescence intensity shown in Table 5 and Figure 4, the mRNA lipid nanoparticles corresponding to compounds 1 and 2 were injected into the tail vein, and the protein expression level was significantly better than that of the comparison compound 1.

1.7 評估脂質顆粒組成物體內尾靜脈注射mRNA肺部表達效率為評估脂質體奈米顆粒將mRNA在肺部有效遞送mRNA並表達相應編碼的蛋白質，以0.5 mg/kg的劑量對6-8週齡雌性BALB/c尾靜脈注射包封有表達螢光素酶的mRNA脂質體奈米顆粒。6小時後，分別向每隻小鼠腹腔注射螢光素酶受質後，取小鼠肺部組織，使用IVIS小動物光學活體成像儀器（PerkinElme），拍攝螢光圖片，並統計螢光強度。螢光強度的高低代表螢光素酶蛋白的表達量高低，即反應脂質體奈米顆粒體內遞送mRNA的效率。每個化合物對應的LNP至少3組重複計算平均螢光值強度，數據見表6和圖5。 1.7 Evaluate the efficiency of mRNA expression in the lungs after tail vein injection of lipid particles In order to evaluate the effective delivery of mRNA in the lung by liposomal nanoparticles and the expression of the corresponding encoded protein, 6-8 weeks old female BALB/c tail vein injected with encapsulated luciferase at a dose of 0.5 mg/kg mRNA liposomal nanoparticles. Six hours later, each mouse was injected intraperitoneally with a luciferase substrate, and the lung tissues of the mice were taken, and fluorescent pictures were taken with an IVIS small animal optical in vivo imaging instrument (PerkinElme), and the fluorescence intensity was counted. The level of fluorescence intensity represents the level of expression of luciferase protein, which reflects the efficiency of delivering mRNA in vivo by liposome nanoparticles. For each compound, at least three groups of LNPs were used to repeatedly calculate the average fluorescence intensity, and the data are shown in Table 6 and Figure 5.

圖5中*即統計學0.01＜P＜0.05，表示組間具有統計學顯著性差異，**即統計學P＜0.01，***即統計學P＜0.001表示極顯著性差異。In Fig. 5, * means statistically 0.01<P<0.05, indicating statistically significant difference between groups, ** means statistically P<0.01, *** means statistically P<0.001 means extremely significant difference.

表6 編號平均螢光強度值化合物4 1.72E+06 對比化合物1 4.74E+05 結論：如表6和圖5所示螢光強度，化合物4對應的mRNA脂質奈米顆粒尾靜脈注射，肺部蛋白表達量顯著優於對比化合物1。 Table 6 serial number Mean Fluorescence Intensity Value Compound 4 1.72E+06 Comparative compound 1 4.74E+05 Conclusion: As shown in the fluorescence intensity in Table 6 and Figure 5, the mRNA lipid nanoparticles corresponding to compound 4 were injected into the tail vein, and the protein expression in the lung was significantly better than that of the comparison compound 1.

無none

圖1：經選定脂質（化合物1-3與對比化合物1和2）遞送後，細胞內mRNA螢光素酶表達量比較。圖2：經選定脂質（化合物1-3與對比化合物1）遞送後，小鼠注射部位mRNA螢光素酶表達量比較。圖3：經選定脂質（化合物1、3與對比化合物1）遞送後，體內肌肉注射生長因子蛋白表達量比較。圖4：經選定脂質（化合物1、2與對比化合物1）遞送後，小鼠全身的mRNA螢光素酶表達量比較。圖5：經選定脂質（化合物4與對比化合物1）遞送後，小鼠肺部的mRNA螢光素酶表達量比較。 Figure 1: Comparison of intracellular mRNA luciferase expression after delivery of selected lipids (compounds 1-3 and comparative compounds 1 and 2). Figure 2: Comparison of mRNA luciferase expression at injection sites in mice after delivery of selected lipids (compounds 1-3 and comparative compound 1). Figure 3: Comparison of protein expression levels of growth factors injected intramuscularly in vivo after delivery of selected lipids (compounds 1, 3 and comparative compound 1). Figure 4: Comparison of mRNA luciferase expression levels in the whole body of mice after delivery of selected lipids (compounds 1, 2 and comparative compound 1). Figure 5: Comparison of mRNA luciferase expression in mouse lungs after delivery of selected lipids (compound 4 and comparative compound 1).

Claims

一種式I所示化合物或其鹽，

其中，M ¹至M ⁶各自獨立地選自鍵、-C(O)O-a1和-OC(O)-a1，a1為與R ¹、R ²、R ³、R ⁴、R ⁵或R ⁶連接的鍵，且M ¹至M ⁶不全為-C(O)O-a1或鍵； R ¹至R ⁶各自獨立地為取代或未被取代的烷基或取代或未被取代的烯基； R ⁷各自獨立地為氫或取代或未被取代的C _1-6烷基； m1至m3各自獨立地選自1、2、3、4、5、6、7和8； n1至n6各自獨立地選自0、1、2、3、4、5、6、7、8、9和10。 A compound represented by formula I or a salt thereof,

如請求項1所述的化合物或其鹽，其中R ¹至R ⁶各自獨立地為未被取代的烷基或未被取代的烯基，較佳C ₄-C ₁₄烷基或C ₄-C ₁₄烯基。 The compound or salt thereof as claimed in claim 1, wherein R ¹ to R ⁶ are each independently unsubstituted alkyl or unsubstituted alkenyl, preferably C ₄ -C ₁₄ alkyl or C ₄ -C ₁₄ alkenyl.

如請求項1或2所述的化合物或其鹽，其中R ¹為C ₄-C ₁₄烷基，較佳C ₅-C ₁₂烷基。 The compound or salt thereof as claimed in claim 1 or 2, wherein R ¹ is C ₄ -C ₁₄ alkyl, preferably C ₅ -C ₁₂ alkyl.

如請求項1至3中任一項所述的化合物或其鹽，其中R ²為C ₄-C ₁₄烷基，較佳C ₅-C ₁₂烷基。 The compound or salt thereof according to any one of claims 1 to 3, wherein R ² is C ₄ -C ₁₄ alkyl, preferably C ₅ -C ₁₂ alkyl.

如請求項1至4中任一項所述的化合物或其鹽，其中R ³為C ₄-C ₁₄烷基，較佳C ₅-C ₁₂烷基；和/或R ⁴為C ₄-C ₁₄烷基，較佳C ₅-C ₁₂烷基；和/或R ⁵為C ₄-C ₁₄烷基，較佳C ₅-C ₁₂烷基；和/或R ⁶為C ₄-C ₁₄烷基，較佳C ₅-C ₁₂烷基。 The compound or salt thereof as claimed in any one of claims 1 to 4, wherein R ³ is C ₄ -C ₁₄ alkyl, preferably C ₅ -C ₁₂ alkyl; and/or R ⁴ is C ₄ -C ₁₄ alkyl, preferably C ₅ -C ₁₂ alkyl; and/or R ⁵ is C ₄ -C ₁₄ alkyl, preferably C ₅ -C ₁₂ alkyl; and/or R ⁶ is C ₄ -C ₁₄ alkane group, preferably C ₅ -C ₁₂ alkyl group.

如請求項1至5中任一項所述的化合物或其鹽，其中R ⁷為氫或視需要取代的C _1-3烷基，較佳氫、甲基或乙基。 The compound or salt thereof as claimed in any one of claims 1 to 5, wherein R ⁷ is hydrogen or optionally substituted C _1-3 alkyl, preferably hydrogen, methyl or ethyl.

如請求項1至6中任一項所述的化合物或其鹽，其中M ¹至M ⁶各自獨立地為-OC(O)-a1，a1為連接R ¹、R ²、R ³、R ⁴、R ⁵或R ⁶的鍵。 The compound or salt thereof according to any one of claims 1 to 6, wherein M ¹ to M ⁶ are each independently -OC(O)-a1, and a1 is the link R ¹ , R ² , R ³ , R ⁴ , R ⁵ or R ⁶ bond.

如請求項1至6中任一項所述的化合物或其鹽，其中M ¹和M ²各自獨立地為-C(O)O-a1，a1為與R ¹或R ²連接的鍵。 The compound or salt thereof as described in any one of claims 1 to 6, wherein M ¹ and M ² are each independently -C(O)O-a1, and a1 is a bond connecting R ¹ or R ² .

如請求項1至6中任一項所述的化合物或其鹽，其中M ¹、M ²、M ³和M ⁴各自獨立地為-C(O)O-a1，a1為與R ¹、R ²、R ³或R ⁴連接的鍵。 The compound or salt thereof according to any one of claims 1 to 6, wherein M ¹ , M ² , M ³ and M ⁴ are each independently -C(O)O-a1, and a1 is the same as R ¹ , R ^2. A bond to which R ³ or R ⁴ is connected.

如請求項1至9中任一項所述的化合物或其鹽，其中m1至m3各自獨立地選自2、3和4。The compound or salt thereof according to any one of claims 1 to 9, wherein m1 to m3 are each independently selected from 2, 3 and 4.

如請求項1至10中任一項所述的化合物或其鹽，其中n1至n6各自獨立地選自6、7、8、9和10。The compound or salt thereof according to any one of claims 1 to 10, wherein n1 to n6 are each independently selected from 6, 7, 8, 9 and 10.

如請求項1或8所述的化合物或其鹽，其中式I所示化合物或其鹽為式II所示化合物或其鹽，

，其中R ¹至R ⁷、m1至m3、n1至n6如請求項1中定義。 The compound or its salt as described in Claim 1 or 8, wherein the compound or its salt shown in Formula I is the compound or its salt shown in Formula II,

, wherein R ¹ to R ⁷ , m1 to m3, and n1 to n6 are as defined in Claim 1.

如請求項1或7所述的化合物或其鹽，其中式I所示化合物或其鹽為式III所示化合物或其鹽，

，其中R ¹至R ⁷、m1至m3、n1至n6如請求項1中定義。 The compound or its salt as described in Claim 1 or 7, wherein the compound or its salt shown in Formula I is the compound or its salt shown in Formula III,

, wherein R ¹ to R ⁷ , m1 to m3, and n1 to n6 are as defined in Claim 1.

如請求項1所述的化合物或其鹽，其中R ¹和R ²各自獨立地選自

、

、

、

和

。 The compound or salt thereof as described in claim 1, wherein R ¹ and R ² are each independently selected from

,

and

.

如請求項1所述的化合物或其鹽，其中該化合物為

、

、

、

、

、

、

、

、

或

。

The compound or salt thereof as described in claim 1, wherein the compound is

,

or

.

一種如請求項1至15中任一項所述的化合物或其鹽的同位素取代物，較佳地，該同位素取代物為氘原子取代物。An isotopic substitution of the compound or salt thereof as described in any one of claims 1 to 15, preferably, the isotopic substitution is a deuterium atom substitution.

一種脂質顆粒，其包含如請求項1至15中任一項所述的化合物或其鹽，或如請求項16所述的同位素取代物，進一步包含活性劑，該活性劑較佳選自免疫刺激性寡核苷酸、siRNA、反義寡核苷酸、mRNA和質粒。A lipid particle comprising the compound or salt thereof as described in any one of claims 1 to 15, or the isotope substitution as described in claim 16, further comprising an active agent, the active agent is preferably selected from immunostimulatory oligonucleotides, siRNA, antisense oligonucleotides, mRNA, and plasmids.

一種醫藥組成物，其包含如請求項17所述的脂質顆粒和藥學上可接受的賦形劑。A pharmaceutical composition, which comprises the lipid particle as described in claim 17 and a pharmaceutically acceptable excipient.

一種如請求項1至15中任一項所述的化合物或其鹽、或如請求項16所述的同位素取代物、或如請求項17所述的脂質顆粒、或如請求項18所述的醫藥組成物在製備用於誘導受試者免疫反應的藥物中的用途。A compound or salt thereof as described in any one of claim items 1 to 15, or an isotope substitution as described in claim item 16, or a lipid particle as described in claim item 17, or as described in claim item 18 Use of the pharmaceutical composition in the preparation of a medicament for inducing an immune response in a subject.

一種如請求項1至15中任一項所述的化合物或其鹽、或如請求項16所述的同位素取代物、或如請求項17所述的脂質顆粒、或如請求項18所述的醫藥組成物在製備用於預防和/或治療與多肽過表達相關的疾病或病症的藥物中的用途。A compound or salt thereof as described in any one of claim items 1 to 15, or an isotope substitution as described in claim item 16, or a lipid particle as described in claim item 17, or as described in claim item 18 Use of the pharmaceutical composition in the preparation of medicines for preventing and/or treating diseases or conditions related to polypeptide overexpression.

一種如請求項1至15任一項所述的化合物或其鹽、或如請求項16所述的同位素取代物、或如請求項17所述的脂質顆粒、或如請求項18所述的醫藥組成物在製備用於預防和/或治療與多肽表達不足相關的疾病或病症的藥物中的用途。A compound or salt thereof as described in any one of claims 1 to 15, or an isotope substitution as described in claim 16, or a lipid particle as described in claim 17, or a medicine as described in claim 18 Use of the composition in the preparation of a medicament for preventing and/or treating a disease or condition associated with underexpression of a polypeptide.