CN115010915A - Nitrogen branched nine-arm polyethylene glycol derivative, preparation method and biological related substance modified by nitrogen branched nine-arm polyethylene glycol derivative - Google Patents

Nitrogen branched nine-arm polyethylene glycol derivative, preparation method and biological related substance modified by nitrogen branched nine-arm polyethylene glycol derivative Download PDF

Info

Publication number
CN115010915A
CN115010915A CN202110245443.7A CN202110245443A CN115010915A CN 115010915 A CN115010915 A CN 115010915A CN 202110245443 A CN202110245443 A CN 202110245443A CN 115010915 A CN115010915 A CN 115010915A
Authority
CN
China
Prior art keywords
group
groups
branched
polyethylene glycol
nitrogen
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN202110245443.7A
Other languages
Chinese (zh)
Other versions
CN115010915B (en
Inventor
翁文桂
刘超
王爱兰
闫策
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
XIAMEN SINOPEG BIOTECH CO Ltd
Original Assignee
XIAMEN SINOPEG BIOTECH CO Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by XIAMEN SINOPEG BIOTECH CO Ltd filed Critical XIAMEN SINOPEG BIOTECH CO Ltd
Priority to CN202110245443.7A priority Critical patent/CN115010915B/en
Publication of CN115010915A publication Critical patent/CN115010915A/en
Application granted granted Critical
Publication of CN115010915B publication Critical patent/CN115010915B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
    • C08G65/00—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule
    • C08G65/02—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule from cyclic ethers by opening of the heterocyclic ring
    • C08G65/26—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule from cyclic ethers by opening of the heterocyclic ring from cyclic ethers and other compounds
    • C08G65/2618—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule from cyclic ethers by opening of the heterocyclic ring from cyclic ethers and other compounds the other compounds containing nitrogen
    • C08G65/2621—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule from cyclic ethers by opening of the heterocyclic ring from cyclic ethers and other compounds the other compounds containing nitrogen containing amine groups
    • C08G65/2624—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule from cyclic ethers by opening of the heterocyclic ring from cyclic ethers and other compounds the other compounds containing nitrogen containing amine groups containing aliphatic amine groups
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
    • A61K47/59—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes
    • A61K47/60—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
    • C08G65/00—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule
    • C08G65/02—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule from cyclic ethers by opening of the heterocyclic ring
    • C08G65/32—Polymers modified by chemical after-treatment
    • C08G65/329—Polymers modified by chemical after-treatment with organic compounds
    • C08G65/331—Polymers modified by chemical after-treatment with organic compounds containing oxygen
    • C08G65/332—Polymers modified by chemical after-treatment with organic compounds containing oxygen containing carboxyl groups, or halides, or esters thereof
    • C08G65/3322—Polymers modified by chemical after-treatment with organic compounds containing oxygen containing carboxyl groups, or halides, or esters thereof acyclic
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
    • C08G65/00—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule
    • C08G65/02—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule from cyclic ethers by opening of the heterocyclic ring
    • C08G65/32—Polymers modified by chemical after-treatment
    • C08G65/329—Polymers modified by chemical after-treatment with organic compounds
    • C08G65/333—Polymers modified by chemical after-treatment with organic compounds containing nitrogen
    • C08G65/33303—Polymers modified by chemical after-treatment with organic compounds containing nitrogen containing amino group
    • C08G65/33306—Polymers modified by chemical after-treatment with organic compounds containing nitrogen containing amino group acyclic
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
    • C08G65/00—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule
    • C08G65/02—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule from cyclic ethers by opening of the heterocyclic ring
    • C08G65/32—Polymers modified by chemical after-treatment
    • C08G65/329—Polymers modified by chemical after-treatment with organic compounds
    • C08G65/333—Polymers modified by chemical after-treatment with organic compounds containing nitrogen
    • C08G65/33303—Polymers modified by chemical after-treatment with organic compounds containing nitrogen containing amino group
    • C08G65/3331—Polymers modified by chemical after-treatment with organic compounds containing nitrogen containing amino group cyclic
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
    • C08G65/00—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule
    • C08G65/02—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule from cyclic ethers by opening of the heterocyclic ring
    • C08G65/32—Polymers modified by chemical after-treatment
    • C08G65/329—Polymers modified by chemical after-treatment with organic compounds
    • C08G65/333—Polymers modified by chemical after-treatment with organic compounds containing nitrogen
    • C08G65/3332—Polymers modified by chemical after-treatment with organic compounds containing nitrogen containing carboxamide group
    • C08G65/33324—Polymers modified by chemical after-treatment with organic compounds containing nitrogen containing carboxamide group acyclic
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
    • C08G65/00—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule
    • C08G65/02—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule from cyclic ethers by opening of the heterocyclic ring
    • C08G65/32—Polymers modified by chemical after-treatment
    • C08G65/329—Polymers modified by chemical after-treatment with organic compounds
    • C08G65/333—Polymers modified by chemical after-treatment with organic compounds containing nitrogen
    • C08G65/3332—Polymers modified by chemical after-treatment with organic compounds containing nitrogen containing carboxamide group
    • C08G65/33327—Polymers modified by chemical after-treatment with organic compounds containing nitrogen containing carboxamide group cyclic
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/50—Improvements relating to the production of bulk chemicals
    • Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Landscapes

  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Polymers & Plastics (AREA)
  • Organic Chemistry (AREA)
  • General Chemical & Material Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Engineering & Computer Science (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Medicinal Preparation (AREA)
  • Polyethers (AREA)

Abstract

The invention discloses a nitrogen-branched nine-arm polyethylene glycol derivative shown in a general formula (1), a preparation method and a biological related substance modified by the nitrogen-branched nine-arm polyethylene glycol derivative. In the invention, the nitrogen-branched nine-arm polyethylene glycol derivative and the biological related substance modified by the nitrogen-branched nine-arm polyethylene glycol derivative are further prepared from nine hydroxyl small molecules or nine functionalized small molecules with basically the same terminal hydroxyl activity, so that the nitrogen-branched nine-arm polyethylene glycol derivative has more advantages in purity and better performance, and the purification difficulty and the preparation cost are reduced. Compared with linear polyethylene glycol and other multi-arm polyethylene glycol, the nitrogen-branched nine-arm polyethylene glycol derivative disclosed by the invention can improve the drug loading rate and the drug activity, and a degradable connecting group is allowed to exist in the connecting group,can be degraded to release medicine and also can improve the drug effect. The invention provides a new idea for constructing a novel nitrogen-branched polyhydroxy initiator molecule and further preparing a nitrogen-branched multi-arm polyethylene glycol derivative.

Description

Nitrogen branched nine-arm polyethylene glycol derivative, preparation method and biological related substance modified by nitrogen branched nine-arm polyethylene glycol derivative
Technical Field
The invention relates to the field of polymer synthesis, in particular to a nitrogen-branched nine-arm polyethylene glycol derivative, a preparation method and a modified biologically-related substance thereof.
Background
Pegylation (PEGylation) is one of the important means for drug modification. The functionalized polyethylene glycol (PEG) can be coupled with drug molecules (including protein drugs and organic small molecule drugs), peptides, saccharides, lipids, oligonucleotides, affinity ligands, cofactors, liposomes, biological materials and the like through covalent bonds by utilizing active groups contained in the functionalized polyethylene glycol (PEG), so that the polyethylene glycol modification of drugs and other biologically relevant substances is realized. The modified drug molecule has many excellent properties of polyethylene glycol (such as hydrophilicity, flexibility, anticoagulation, etc.). Meanwhile, due to the steric exclusion effect, the drug modified by the polyethylene glycol avoids the filtration of glomeruli and biological reactions such as immunoreaction, so that the drug has longer half-life in blood than the unmodified drug. For example: greenwald et al (J.org.chem.1995,331-336) modify paclitaxel by means of coupling with polyethylene glycol to increase its aqueous solubility.
Since 1995, Monfardini grafted two linear methoxypolyethylene glycols to two amino groups of lysine to obtain two-armed branched (V-type) polyethylene glycols, activated the carboxyl group of lysine to succinimide-active esters, and used for protein modification studies (Bioconjugate chem.1995,6,62-69), this method was generalized to the most popular method for preparing monofunctional branched polyethylene glycols and their drug derivatives, and has been used in three commercial drugs. Compared with linear polyethylene glycol with the same molecular weight, the polyethylene glycol with the branched chain can form an umbrella-shaped protective layer on the surface layer of the medicine due to the special molecular form, so that the steric hindrance around the medicine molecules is increased, the attack of other macromolecular substances in vivo on the medicine can be more effectively prevented compared with the linear polyethylene glycol, the degree of inactivation or enzymatic hydrolysis of the medicine in vivo is reduced, and the action time of the medicine in vivo is prolonged.
In addition to linear monofunctional and linear bifunctional polyethylene glycol products, multi-arm polyethylene glycols such as three-arm polyethylene glycol, four-arm polyethylene glycol, six-arm polyethylene glycol, eight-arm polyethylene glycol, etc. begin to occupy a place in the market due to their advantages in structure and drug loading. Especially for small molecule drugs, the clinical application of the small molecule drugs is greatly limited due to low solubility and large toxic and side effects. And the traditional linear structure is adopted, and the single functionalization or the double functionalization is adopted, so that the solubility is improved, the toxic and side effects are reduced, and meanwhile, the drug activity is greatly reduced because the drug molecules are embedded by the PEG chain. Compared with polyethylene glycol derivatives with linear structures and multi-arm branched structures, the solubility can be improved, the toxic and side effects can be reduced, the drug loading capacity can be improved, and the drug activity can be kept high. In addition, the system viscosity of the multi-arm structure is greatly reduced, and better pharmacokinetics is favorably obtained. At present, two cases of small molecule drugs modified by four-arm polyethylene glycol enter clinical II and III stages.
Although multi-arm structures such as three-arm, four-arm and six-arm structures exist in the prior art, the drug loading capacity is limited, and initiator molecules with central structures are often asymmetric in polyhydroxy micromolecules, namely, the activities of hydroxyl groups at the tail end of each side of the catalyst are not completely the same, so that the reaction activities are different when ethylene oxide polymerization is initiated, the chain lengths of polyethylene glycol are different, the molecular weight uniformity is not ideal, and the molecular weight distribution is wide. Hexavalent center structure hexahydroxy small molecules (HOCH), such as tetrapolyglycerol branched polyethylene glycol 2 (HO)CHCH 2 OCH[CH 2 OCH 2 CH(OH)CH 2 OH] 2 ) The asymmetry is that the atom spacing from each terminal hydroxyl group to the branching center is different, resulting in differences in their reactivity.
In view of the above background, it is necessary to develop an initiator molecule having symmetrical branched terminal functional groups and substantially the same reactivity and containing a plurality of hydroxyl groups or a plurality of other functional groups, and further to prepare a nitrogen-branched nona-armed polyethylene glycol derivative having a uniform chain length and a uniform molecular weight.
Disclosure of Invention
The invention provides a nitrogen-branched nine-arm polyethylene glycol derivative, a preparation method and a biological related substance modified by the nitrogen-branched nine-arm polyethylene glycol derivative against the background.
The purpose of the invention is realized by the following technical scheme:
a nitrogen-branched nine-arm polyethylene glycol derivative has a structure shown in a general formula (1):
Figure BDA0002963906360000021
wherein, N is a nitrogen atom trivalent branching center;
U 0 is a tetravalent radical, 3U's in the same molecule 0 The same; a trivalent branching center of nitrogen atoms and 3U 0 Together form a nine-valent branched structure;
L 2 is absent or L 2 Is a divalent linking group; in the same molecule, 9L 2 Are the same or different from each other;
PEG is a polyethylene glycol block taking ethylene oxide as a repeating unit, and both ends of the PEG are oxygen radicals;
F G is a hydrogen atom or a terminal functional group; when F is present G Containing at least one terminal functional group R when not a hydrogen atom 01 (ii) a Wherein R is 01 Is a functional group capable of interacting with biologically relevant substances.
The invention also provides a preparation method of the nitrogen-branched nine-arm polyethylene glycol derivative, which comprises the following steps:
step one, adopting nine hydroxyl small molecules containing nine hydroxyl groups
Figure BDA0002963906360000022
The initiator system of (1); wherein, deprotonation of nine bare hydroxyl groups forms nonaxonium anions
Figure BDA0002963906360000023
(also note as
Figure BDA0002963906360000024
Or
Figure BDA0002963906360000025
) In the vaginaStable existence under the condition of ionic polymerization;
initiating ethylene oxide polymerization;
step three, after the reaction is finished, adding a proton source into the intermediate product system with nine polyethylene glycol chains obtained in the step two to obtain hydroxyl-terminated nitrogen-branched nine-arm polyethylene glycol;
step four, functionalizing the tail end of the nitrogen-branched nine-arm polyethylene glycol to obtain the nitrogen-branched nine-arm polyethylene glycol derivative, wherein the terminal functionalization is selected from any one of terminal linear functionalization and terminal branched functionalization; when g is 0, terminal linear functionalization is performed, and when g is 1, terminal branched functionalization is performed.
The invention also provides a preparation method of the nitrogen-branched nine-arm polyethylene glycol derivative, and relates to 1 nine functionalized small molecule containing nine same functional groups
Figure BDA0002963906360000031
Reacting with 9 linear double-end functionalized PEG derivative bilPEG molecules in a coupling reaction process to obtain a nitrogen-branched nine-arm polyethylene glycol derivative; wherein bilPEG is monodisperse or polydisperse; wherein functional groups at both ends of bilPEG may be the same or different; wherein, F 1 Contains a reactive functional group capable of reacting with a terminal functional group in the biliPEG to form a divalent linking group L 2 ；
The functional group at the other end of the bilPEG is the same as or different from the target structure; when different, the method further comprises branching the nitrogen into nine-arm polyethylene glycol
Figure BDA0002963906360000032
The terminal functionalization process of (a), the terminal functionalization being selected from any one of terminal linear functionalization and terminal branched functionalization; when g is 0, terminal linear functionalization is performed, and when g is 1, terminal branched functionalization is performed.
The invention also provides a nitrogen-branched nine-arm polyethylene glycol derivative modified biologically-relevant substance, which has the following structural general formula:
Figure BDA0002963906360000033
wherein N is a nitrogen atom trivalent branching center;
U 0 being a tetravalent radical, 3U's in the same molecule 0 The same; n and three U 0 Together form a nine-valent branched structure;
L 2 is absent or L 2 Is a divalent linking group; in the same molecule, nine L 2 Identical or different, preferably identical, from one another;
PEG is a polyethylene glycol block taking ethylene oxide as a repeating unit, and both ends of the PEG are oxygen radicals;
k is the number of EF in a single functionalized end and is selected from 1 or 2-250;
in a single functionalized PEG chain, g is 0 or 1; g is a terminal branching group selected from a trivalent or higher valent connecting group which connects the PEG chain segment with a terminal functional group; l is 0 Is a divalent linking group which connects the PEG chain segment with the terminal branching group G;
when g is 0, k is 1, L 0 G is not present;
when G is 1, G is present, L 0 May be present or absent, k is 2-250;
EF may be expressed as ED (structure is
Figure BDA0002963906360000041
) Or EF 1 (structure is
Figure BDA0002963906360000042
) And D is not equal to E 01 (ii) a Wherein q and q are 1 Each independently is 0 or 1; z 1 、Z 2 Each independently is a divalent linking group; wherein D is a residue formed by reacting the modified biological related substance with the nitrogen-branched nine-arm polyethylene glycol derivative; e 01 Is selected from R 01 Protected R 01 Deprotected R 01 Or blocked R 01 ；R 01 Is a reactive group capable of reacting with a biologically-relevant substance; l is nitrogen branched nine-armA linking group formed after a reactive group in the polyethylene glycol derivative reacts with a biologically relevant substance; wherein the number of D at the end of one branch chain is denoted by k D ，0≤k D K is not more than k, k of each branched chain in the same molecule D Each independently the same or different, and the sum of the numbers of D (N) in any one of the nitrogen-branched nine-arm polyethylene glycol derivative molecules D ) At least 1, preferably at least 9; when G is 1, then G- (EF) k Can be expressed as
Figure BDA0002963906360000043
The biologically-relevant substance preferably has a plurality of reaction sites, and the same biologically-relevant substance and the same R 01 The residues D obtained by the reaction may be the same or different;
the biologically-relevant substance modified by the nitrogen-branched nine-arm polyethylene glycol derivative can exist stably or can be degraded; in the same molecule, U 0 、L 2 、L 0 、G、(Z 2 ) q -(Z 1 ) q1 、(Z 2 ) q Any of L, or any of the linkages formed with adjacent groups, may each independently be stable or degradable.
Compared with the prior art, the invention has the following beneficial effects:
(1) the nitrogen branched nine-arm polyethylene glycol can be obtained by adopting symmetric nine-hydroxyl micromolecules as initiator molecules through polymerization reaction. "symmetrical type" means that the nine terminal hydroxyl groups of the nine-hydroxyl small molecule have almost the same activity, and the nine-hydroxyl small molecule can be divided into three functional small molecules with one residue as the trivalent branching center of nitrogen atom and 3 residues as U 0 The tetrafunctional small molecule is obtained by coupling reaction. Wherein, the residue can be used as a tri-functional micromolecule of a nitrogen atom trivalent branching center, and the tri-functional micromolecule contains three same functional groups; the residue may be U 0 The tetrafunctional small molecule of (2) contains three or four identical functional groups and consists of U 0 At least three ends of the branched core are completely the same, when the residue can be U 0 When the tetrafunctional small molecule of (a) contains only three identical functional groups, different onesThe end of each functional group is connected with a trifunctional small molecule of which the residue can be a nitrogen atom trivalent branching center. The nine hydroxyl groups of the nine hydroxyl small molecule prepared in the way are almost the same in activity, and the nine hydroxyl groups are respectively connected with U 0 The atom intervals of the branched cores are the same, the nitrogen-branched nine-arm polyethylene glycol derivative further prepared by using the nine-hydroxyl micromolecule as an initiator molecule has more advantages in purity, the molecular weight and the distribution thereof are more accurately controlled in the product polymerization process, the product has a single structure, the structure of other multi-arm products cannot be generated, the performance is better, the purification difficulty is reduced, the using amount of organic reagents in purification can be reduced, the cost is reduced, the environment is more green and environment-friendly, and compared with the prior art in which the terminal hydroxyl asymmetric nine-hydroxyl micromolecule is used as an initiator, the performance of the obtained product is better.
(2) Compared with linear polyethylene glycol, three-arm polyethylene glycol, four-arm polyethylene glycol and six-arm polyethylene glycol, the nitrogen branched nine-arm structure increases the number of active groups and improves the drug loading rate.
(3) Trivalent and above trivalent terminal branching groups can be introduced between the functional groups and the PEG arms, so that the number of active groups in the polyethylene glycol is further increased, the drug loading capacity is improved to a great extent, and the drug effect can be further improved.
(4) In addition to the introduction of nine polyethylene glycol arms by direct initiation of ethylene oxide polymerization using a nine hydroxyl small molecule initiator, nine linear PEG arms can be coupled to the end of a nine functionalized small molecule by a coupling method. The nine-functional small molecule contains 9 identical functional groups selected from suitable reactive groups in class a-class H of the present invention. The nine-functional small molecule can be a tri-functional small molecule containing three same functional groups, the residue of which can be a nitrogen atom trivalent branching center, and the 3 residues of which can be U 0 Is obtained by coupling reaction of at least four functionalized small molecules containing three same functional groups and is composed of U 0 At least three ends led out from the branched core are completely identical. When the residue may be U 0 When the tetrafunctional small molecule only contains three same functional groups, different functional groups are connected with the trifunctional small molecule of which the residue can be a nitrogen atom trivalent branching center, so that the tetrafunctional small molecule hasThe activities of 9 terminal functional groups of the nine functionalized micromolecules are almost the same, accurate molecular weight and narrower molecular weight distribution can be given to the nitrogen branched nine-arm polyethylene glycol during the subsequent coupling reaction with the linear polyethylene glycol, and the difficulty of purification and separation can be reduced, the using amount of organic reagents is reduced, the cost is reduced, and the method is more green and environment-friendly in the subsequent purification process. The molecular weight of the coupling product with high molecular weight is more uniform and the polydispersity index (PDI) is lower along with the redistribution of the weight percentage of the molecular weight; for higher molecular weights of the nitrogen-branched nine-arm structure, a narrower molecular weight distribution can be achieved by reducing the PDI of the single-chain feedstock.
(5) Three branched polyethylene glycol molecules V-PEG containing three same branched PEG chain segments can also be combined by a coupling method 3 Coupling to the ends of a small trifunctional molecule whose residue may be a trivalent branching center for the nitrogen atom renders the coupling product more uniform in molecular weight.
(6) The product with single molecular weight of PDI-1 can be obtained, and the corresponding nitrogen branched nine-arm polyethylene glycol derivative and the biological related substances modified by the same have determined structure and molecular weight, and have higher degree of standardized control and production.
(7) Nitrogen-branched nine-armed polyethylene glycol derivatives, U, of the invention 0 、L 2 、L 0 、Z 1 、Z 2 Any one of them or any one of the linkers with the adjacent heteroatom allows the presence of a degradable linker, which can degrade under specific circumstances; when only U is present 0 When degradable, 1-3 three-arm branched PEGs can be degraded; when only L is present 2 When degradable, 1-9 linear PEG chains can be degraded; when is- (Z) 2 ) q -(Z 1 ) q1 When the position of (E) is degraded, the drug molecule and the polyethylene glycol structure are separated, so that the active site of the drug molecule is exposed to the maximum extent, and the drug molecule can approach the unmodified state to the maximum extent.
(8) In the biologically-relevant substance modified by the nitrogen-branched nine-arm polyethylene glycol derivative, the existence of the degradable connecting group is allowed between the functional group (F) and the polyethylene glycol chain, so that the substance is allowed to be degraded and released in a specific environment, the tissue distribution of the medicament is improved, the accumulation at a focus part is increased, and the medicament effect is improved.
(9) The nitrogen branched nine-arm polyethylene glycol can also be used as a cross-linking agent to be applied to the preparation of gels, and the gels can be used as an adhesive, an anti-seepage agent, an anti-adhesion agent, a hemostatic material and the like in medical instruments.
Detailed Description
Description of the terms
In the present invention, the oxyethylene unit as the repeating unit of the polyethylene glycol component is also referred to as EO unit, the number of repeating units is also referred to as EO unit number, the average number of repeating units is also referred to as EO unit average number, and the number average number is preferable.
The terms related to the invention are mostly described in documents CN104530415A, CN104530417A, CN104877127A, WO/2016/206540A, US15/738761, CN106967213A, CN201710125672.9, CN201710126727.8, CN201911013365.7, CN202010209573.0 and various cited documents, including but not limited to the term interpretation part of the specification; the explanations of terms, examples of related structures, and preferred embodiments described in the above documents and the cited documents are incorporated by reference into the present invention and will not be described in detail herein.
The description in the cited documents of the present invention is different from the description of the present invention, and the present invention is subject to the standard; this principle is addressed to all citations throughout the specification.
In general terms referred to in the literature include, but are not limited to, hydrocarbons, aliphatic hydrocarbons, aromatic hydrocarbons, saturated hydrocarbons, alkanes, saturated aliphatic hydrocarbons, unsaturated aliphatic hydrocarbons, alkenes, alkynes, dienes, open chain hydrocarbons, straight chain structures (without pendant groups), straight chain hydrocarbons, straight chain aliphatic hydrocarbons, branched chain structures (with pendant groups), branched chain aliphatic hydrocarbons, cyclic structures, dendritic structures, comb structures, hyperbranched structures, ring atoms, ring backbones, hydrocarbon-derived rings, carbocyclics, alicyclics, saturated alicyclic hydrocarbons (cycloalkanes), unsaturated alicyclics, unsaturated alicyclic hydrocarbons, cycloalkenes, cycloalkynes, cyclodioles, benzene rings, aromatic rings, fused rings, structural units constituting ring backbones, classes with or without nested cyclic structures, heteroatoms, heterocyclic rings, Aliphatic heterocycles, aromatic heterocycles, heteroalicyclic rings, heteroaromatic rings (heterocyclic rings), oxa-aza-, thia-, phosph-, number of heteroatoms, oxa-aza-, thia-, the position of the heteroatoms, number of ring structures, monocyclic, polycyclic, monocyclic compounds, polycyclic compounds, number of rings, bicyclic, tricyclic, tetracyclic, linkage between ring structures, spiro-, bridged-, any two-linked rings of polycyclic, heteromonocyclic (mono-heterocyclic), polycyclic, spiro-, bridged-, hetero-fused-, fused-aromatic (fused-fused), fused-aromatic (fused-conjugated-heterocyclic), aromatic-fused-heterocyclic (arylheterocyclic), benzo-heterocyclic, fused-aromatic, monocyclic hydrocarbon, polycyclic hydrocarbon, fused-cyclic hydrocarbon, fused-hydrocarbon, heterocyclic-substituted hydrocarbon, heterocyclic-substituted hydrocarbon, heterocyclic-substituted hydrocarbon, heterocyclic, the terms of fused aromatic hydrocarbon, saturated cyclic hydrocarbon (cycloalkane), unsaturated cyclic hydrocarbon, heterohydrocarbon, open-chain heterohydrocarbon, heterocyclic hydrocarbon, aliphatic heterohydrocarbon, aromatic heterohydrocarbon, aliphatic heterocyclic hydrocarbon, aliphatic open-chain heterohydrocarbon, saturated aliphatic heterohydrocarbon (heteroalkane), hetero-aromatic hydrocarbon, fused heterohydrocarbon, fused heterocyclic hydrocarbon, aromatic fused heterocyclic hydrocarbon, heteroaromatic alkane, etc., and the related structures are exemplified and preferred.
Terms referred to in the cited documents also include, but are not limited to: a group, a residue, a valence of the group, a monovalent group, a divalent group, a trivalent group, a tetravalent group, … …, a hundredth group, a linker, an oxy group, a thio group, a hydrocarbon group, a monovalent hydrocarbon group, a divalent hydrocarbon (hydrocarbylene) group, a trivalent hydrocarbon group, a "substituted", a substituent atom, a substituent, a hydrocarbon substituent, a heteroatom-containing group, a heterocarbon group, a heteroatom-containing substituent, an acyl group, a carbonyl group, a non-carbonyl group, a hydrocarbyloxy group, a hydrocarbylthio group, an acyloxy (acyloxy) group, an oxyacyl group, an aminoacyl group, an acylamino group, a substituted hydrocarbon group, a hydrocarbon-substituted hydrocarbon group (still belonging to the group), a saturated hydrocarbon (alkyl), an unsaturated hydrocarbon group, an alkenyl group, an alkynyl group, a dienyl group, an alkenylhydrocarbon group, an alkynyl hydrocarbon group, a dienyl group, an open-chain hydrocarbon group, a straight-chain hydrocarbon group, a branched-chain hydrocarbon group, a cyclic hydrocarbon group, an aliphatic hydrocarbon group, an alicyclic hydrocarbon group, a heterocyclic group, a compound, cycloalkyl groups, unsaturated alicyclic groups, monocyclic alkyl groups, polycyclic alkyl groups, aryl groups, arylalkyl groups, heteroalkyl groups, open-chain heteroalkyl groups, alicyclic hydrocarbon groups, heterocycloalkyl groups, heterocycle-substituted hydrocarbon groups, heteroaromatic groups, heteroaryl groups, heteroarylalkyl groups, fused-ring hydrocarbon groups, fused-ring aryl groups, fused-ring heteroaryl groups, hetero-fused-ring hydrocarbon groups, oxahydrocarbon groups, aza-hydrocarbon groups, thiahydrocarbon groups, phosphane groups, monohetero-hydrocarbon groups, hetero-hydrocarbon groups, polyheterohetero-hydrocarbon groups, alkylene groups, sources of alkylene groups, alkylene groups derived from unsaturated aliphatic hydrocarbons, cycloalkylene groups, alicyclic hydrocarbon groups, arylene groups, cyclic structures, and substituents ps, The term "substituted" or "unsubstituted" alkylene groups may or may not contain substituents or side groups, two positions in the alkylene group to which other groups are attached, protecting groups, thiol protecting groups, alkynyl protecting groups, hydroxyl protecting groups, amino groups, divalent linking groups, and the like. Wherein the substituent atom forms a substituent consisting of only one heteroatom. Wherein alkenyl, alkynyl, dienyl, alkenyl, alkynyl, dialkenyl are defined as follows: unsaturated hydrocarbons lose hydrogen atoms to form radicals such as alkenyl (also known as alkenyl), alkynyl (also known as alkynyl), dialkenyl, and the like. Unsaturated hydrocarbons are hydrocarbon groups formed by losing hydrogen atoms on unsaturated carbons, such as 1-alkenyl, 1-alkynyl, 1-dienyl, and the like, such as propenyl, propynyl, as examples. The unsaturated hydrocarbon group formed by losing a hydrogen atom on a saturated carbon is, depending on the unsaturated bond, for example, an alkenyl hydrocarbon group, an alkynyl hydrocarbon group, an alkadienyl hydrocarbon group and the like, specifically, an allyl group (2-propenyl group), a propargyl group (2-propynyl group).
Terms referred to in the cited documents also include, but are not limited to: a branching center, a branching structure, a triple-branching structure, a quadruple-branching structure, a cyclic structure (including, but not limited to, an aliphatic ring, an aromatic ring, a sugar ring, a condensed ring, a polymer ring, etc.), "stably present", "degradable", "cyclic monosaccharide", "polydisperse", and "monodisperse", a trivalent group and its illustration, a tetravalent group and its illustration, a pentavalent group and its illustration, a hexavalent group and its illustration, a heptavalent group and its illustration, an octavalent group and its illustration, a higher valent group and its illustration, a trivalent group and its illustration in which a part other than the trivalent nuclear structure does not include a heteroatom, a trivalent group and its illustration in which a part other than the trivalent nuclear structure includes a heteroatom, a trivalent branching structure, a tetravalent group and its illustration in which a part other than the tetravalent nuclear structure does not include a heteroatom, a tetravalent group and its illustration in which a part other than the tetravalent nuclear structure includes a heteroatom, a tetravalent group and its illustration in which a part other than the tetravalent core structure includes a heteroatom, Combinations among groups, combinations among low-valent groups (comb combinations, tree combinations, branched combinations, hyperbranched combinations, cyclic combinations, etc.), comb combinations, tree combinations, branched combinations, hyperbranched combinations, cyclic combinations, the same structure type, examples of functional groups and their protected forms, reactive groups, protecting groups for functional groups, protected reactive groups, divalent linking groups that can exist stably, degradable, stable, degradable, divalent linking groups that can exist stably, degradable polyvalent groups, targeting factors, targeting molecules, photosensitive groups (covering fluorescent groups), fluorescent substances, heterofunctional groups (pair of heterofunctional groups that can exist simultaneously), small molecules containing two same or different functional groups, and the like, The definition, description, relevant examples and relevant citations of the hetero-functionalized small molecule (htriSM) containing a trivalent nuclear structure, a terminal functionalization method (linear functionalization and branched functionalization) and raw materials thereof, a coupling reaction and examples thereof, a covalent linking group generated by the coupling reaction, a biological related substance, a reaction between a polyethylene glycol derivative and the biological related substance, site-specific modification, medicine, application fields of small molecule medicine and the like. The preferred embodiments referred to above are also included in the scope of the present invention.
In particular, the structures of CN201710126727.8 and the cited documents relating to the protection and deprotection of functional groups, especially "protection and deprotection of functional groups in reaction raw materials and intermediates" are also included in the present invention. Including, but not limited to, commonly used protecting groups and corresponding protecting and deprotecting methods, thiol protecting groups and protected thiols, amino protecting groups and protected amino groups, hydroxyl protecting groups and protected hydroxyl groups, alkynyl protecting groups and protected alkynyls, aldehyde protecting groups, carboxyl protecting groups, protecting groups of aminocarboxylic acids, protecting groups of natural amino acids, protecting and deprotecting methods of aminocarboxylic acids, protecting and deprotecting methods of natural amino acids (e.g., protecting and deprotecting of amino, carboxyl, amido, thiol, guanidino, β, γ -carboxyl, imidazolyl, ω -amino, β, γ -amide, indolyl, etc.), selective protecting and deprotecting, and the like.
The contents of the document CN201710126727.8 and the cited documents on alkylation reaction and alkylation method are also incorporated into the present invention, including but not limited to three types of substitution, addition and condensation.
Some terms are briefly summarized as follows.
In the present invention, a "group" contains at least 1 atom and refers to a radical formed by a compound having one or more atoms removed. In contrast to compounds, the radicals formed after a compound has lost part of the radical are also referred to as residues. The "group" in the present invention is not a hydrogen atom. The valence of the group is not particularly limited, and may be classified into monovalent group, divalent group, trivalent group, tetravalent group, … …, hundredth-valent group, and the like, as examples. Monovalent groups are exemplified by phenyl, chloro, aldehyde, methoxy, hydroxy, and ethyl groups. With respect to the valence of a group, "multivalent" means that the valence is at least 3.
"group" means the remaining portion of an organic substance after the loss of an atom or group of atoms, the valence being at least 1 and the number of atoms being at least 1. Wherein, the groups with the valence of more than or equal to 2 are collectively called connecting groups. The linking group may also contain only one atom, such as oxy, thio. "substituent" refers to a group in a compound or group that replaces a hydrogen atom. A "substituent" can be an atom (a substituent atom) or a group of two or more atoms (a substituent group). The substituted atoms, substituents, and substituents in the above cited references and in the respective cited references are included in the scope of the "substituents" of the present invention.
In the present invention, alkenyl, alkynyl, dienyl and aryl groups have the same concept as alkenyl, alkynyl, dialkenyl and aryl groups, respectively. And alkenylhydrocarbyl, alkynylalkyl, alkadienylalkyl, arylhydrocarbyl are each composed of "alkenyl + hydrocarbylene," alkynyl + hydrocarbylene, "" alkadienyl + hydrocarbylene, "" aryl + hydrocarbylene. Unsaturated hydrocarbons such as alkenes, alkynes, dienes, and arenes lose a hydrogen atom to form alkenyl groups or alkenyl groups (alkenyl groups), alkynyl groups or alkynyl groups (alkynyl groups), dienyl groups or dienyl groups (dienyl groups), aryl groups (aryl groups), or aryl groups (aryl hydrocarbon groups), respectively. Alkenyl, alkynyl, dienyl, aryl, and the like, with alkylene groups constitute alkenylhydrocarbyl, alkynylalkyl, dienylalkyl, arylalkyl, and the like, while hydrocarbyl groups with alkenylene, alkynylene, dienylene, arylene, and the like, constitute hydrocarbylalkenyl, hydrocarbylalkynyl, hydrocarbyldienyl, hydrocarbylaryl, and the like, respectively. In the case where the present invention is not particularly specified, alkenyl-hydrocarbyl groups (alkenyl-hydrocarbyl groups), alkynyl-hydrocarbyl groups (alkynyl-hydrocarbyl groups), dienyl-hydrocarbyl groups (dienyl-hydrocarbyl groups), and the like include, in particular, those in which the free end group is not located at an unsaturated bond, specifically, allyl (2-propenyl) or propargyl (2-propynyl); the hydrocarbyl-alkenyl groups, hydrocarbyl-alkynyl groups, hydrocarbyl-dienyl groups, and the like particularly include those in which the free terminal group is located on an unsaturated bond, and specific examples thereof include allyl (3-propenyl) and propargyl (3-propynyl).
Monovalent radicals are exemplified. The radicals of aromatic hydrocarbons which are formed by losing one hydrogen atom form aryl or arylalkyl radicals. Arenes are aryl-hydrocarbyl groups formed by the loss of hydrogen atoms from non-aromatic rings. Aryl groups and alkylene groups constitute arylalkyl groups, and alkyl groups and arylene groups constitute alkylaryl groups. Where not specifically stated in the present invention, arylalkyl specifically includes the case where the free end group is not located on an aromatic ring, and alkylaryl specifically includes the case where the free end group is located on an aromatic ring. Arylalkanes lose hydrogen atoms from non-aromatic rings to form arylalkyl groups (aryl groups or arylalkyl groups). Aralkyl groups also fall within the category of aryl, arenyl, and arylalkyl groups. By way of example, most typical aryl groups are phenyl, phenylene, and most typical arylalkyl groups are benzyl.
For simplicity, the range of carbon atoms in a group of the present invention may also be indicated in subscript form at the position of the subscript of C, indicating the number of carbon atoms the group has, e.g. C 1-10 Denotes "having 1 to 10 carbon atoms", C 3-20 Means "having 3 to 20 carbon atoms". "substituted C 3-20 Hydrocarbyl "means C 3-20 A group obtained by substituting a hydrogen atom of a hydrocarbon group. "C 3-20 The substituted hydrocarbon group "means a group having 3 to 20 carbon atoms in the group obtained by substituting the hydrogen atom of the hydrocarbon group. Also for example, when a group can be selected from C 1-10 When hydrocarbyl, it may be selected from hydrocarbyl groups having any one of the number of carbon atoms in the range indicated by the subscript, i.e., equivalent to and may be selected from C 1 、C 2 、C 3 、C 4 、C 5 、C 6 、 C 7 、C 8 、C 9 、C 10 Any of hydrocarbon groups.
The above definitions of integer numerical intervals consisting of integers are not limited to subscripts of carbon atoms, but also apply to subscript intervals of other atoms, and also apply to any other integer numerical intervals in the form of subscripts or non-subscripts, including numerical intervals marked by dashes (e.g., 1 to 6) and numerical intervals marked by wavy lines (e.g., 2 to 250), but not to numerical intervals consisting of non-integers. In the present invention, unless otherwise specified, the integer intervals marked as intervals may represent the group of all integers within the range of the interval, and the range includes both endpoints. The integer range is 2-12 and represents 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12. For another example, the number of oxyethylene units in a PEG segment is selected from 4 to 20, meaning that the number of oxyethylene units can be selected from 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20. The range of the non-integer values is, for example, 22 to 100 as the average number of EO units, and the range is not limited to the integer in the range, and may be any non-integer.
In the present invention, two or more objects "are independently preferred" and, when there are preferences in multiple stages, there is no requirement that they are all selected from the same group of preferences in the same stage, but one may be a wide-range preference, one may be a narrow-range preference, one may be a maximum range, and the other may be any preference, and may also be selected from the same group of preferences. For example, "R 8 、R 9 、R 10 、R 11 、R 12 The number of carbon atoms is preferably 1 to 20, more preferably 1 to 10 ", and may be 1 to 20, 1 to 10, or 1 to 20, and 1 to 10 for some, and 1 to 10 for others. Even if the same class or preference is given, it is not intended that the structures of the two entities be completely identical, for example A, B each independently selected from alkyl, cycloalkyl, aryl, aralkyl, and may be a methyl group and B an ethyl group (both alkyl groups), or a butyl group and B a benzyl group (one alkyl group and one aralkyl group).
In the present invention, the divalent linking group such as alkylene, arylene, amide bond and the like is not particularly limited, and any of the two linking ends may be selected when other groups are linked, for example, in A-CH 2 CH 2 -and-CH 2 When an amide bond is used as a divalent linking group between-B, it may be A-CH 2 CH 2 -C(＝O)NH-CH 2 -B or A-CH 2 CH 2 -NHC(＝O)-CH 2 -B。
In the structural formula of the partial group of the present invention, one or more free ends are marked with asterisks as oriented attachment ends. For example for U 0 The asterisk end points to the nitrogen atom trivalent branching center, and three identical non-asterisk ends point to L respectively 2 E.g. U 0 Is composed of
Figure BDA0002963906360000081
When the asterisk end points to the nitrogen atom trivalent branching center, three identical non-asterisk ends point to three L 2 。
When the structure concerned has an isomer, any of the isomers may be used unless otherwise specified. For example, a cis-isomer or trans-isomer may be present in a structure; when the optical rotation property exists, the optical rotation property can be left-handed rotation or right-handed rotation. As for the alkyl group, a hydrocarbon group formed by an alkane which has lost one or more hydrogen atoms at any position is referred to, and in the case where it is not particularly specified, a monovalent alkyl group is generally referred to. Specifically, for example, propyl means any of n-propyl and isopropyl, and propylene means any of 1, 3-propylene, 1, 2-propylene and isopropylene.
In the formula, when the terminal group of the linking group is easily confused with the substituent contained in the linking group, as in the formula
Figure BDA0002963906360000082
In (1), adopt
Figure BDA0002963906360000083
To mark the position of the divalent linking group to which the other group is attached, both of which represent-CH (CH) 2 CH 2 CH 3 ) 2 -、-CH 2 CH 2 CH(CH 3 ) 2 -CH 2 CH 2 -. When no ambiguity occurs, it may not be marked specifically, e.g. structural formula marked as phenylene structure
Figure BDA0002963906360000084
Are respectively equivalent to
Figure BDA0002963906360000085
In the present invention, the ring structure is represented by a circle, and the ring structure is labeled differently according to the characteristics of the ring structure. For example,
Figure BDA0002963906360000086
represents an arbitrary cyclic structure;
Figure BDA0002963906360000087
represents an aliphatic cyclic structure and does not contain any aromatic or heteroaromatic ring, also known as an aliphatic ring;
Figure BDA0002963906360000088
an aromatic cyclic structure containing at least one aromatic or heteroaromatic ring, also called an aromatic ring (may be an all-carbocyclic ring or may contain a heteroatom);
Figure BDA0002963906360000091
represents a skeleton of a saccharide or saccharide derivative having a cyclic monosaccharide skeleton, also referred to as a saccharide ring;
Figure BDA0002963906360000092
a ring having a chemical bond such as an amide bond, an ester bond, an imide, or an acid anhydride in the ring is referred to as a condensed ring;
Figure BDA0002963906360000093
a cyclic backbone of water-soluble polymers, also known as polymeric rings; the molecular weight of the water-soluble polymer is not particularly limited herein.
By way of example, such as
Figure BDA0002963906360000094
Figure BDA0002963906360000095
Respectively represent a ring structure containing nitrogen atoms, carbon-carbon double bonds, azo groups, carbon-carbon triple bonds, disulfide bonds, conjugated diene bonds, anhydride units, imide bonds and aromatic triazole units.
The definition of the above-mentioned various ring structures also includes, but is not limited to, the lower structures, the corresponding examples and preferred embodiments of the structures, etc. disclosed and cited in the above cited documents and the respective cited documents. For example:
Alicyclic rings (aliphatic rings) include alicyclic and alicyclic rings. Alicyclic rings (alicyclic rings) refer to carbon rings derived from aliphatic hydrocarbon groups, and are all-carbon alicyclic rings. In the present invention, aliphatic-derived heterocycles (alicyclic-derived heterocycles) refer to heterocycles in which a carbon atom of a ring is replaced with a heteroatom. By way of example, aliphatic heterocyclic hydrocarbons are meant heterocyclic hydrocarbons derived from alicyclic hydrocarbons, such as 3-oxetane, 1, 4-dioxan. Examples of the aliphatic ring include cyclopropane, oxirane, aziridine, cyclobutane, cyclobutene, squaric acid, cyclobutanedione, hemisquaric acid, cyclopentane, cyclopentadiene, tetrahydrofuran, pyrrolidine, thiazolidine, dihydroisoxazole, oxazolidine, cyclohexane, cyclohexene, tetrahydropyran, piperidine, 1, 4-dioxane, norbornane, norbornene, norbornadiene, 1,4, 7-triazacyclononane, cyclen, etc., and it is to be noted that a ring having weak aromaticity such as furan, thiophene, pyrrole, imidazole, oxazole, isoxazole, thiazole, isothiazole, pyrazole, etc. is also included in the aliphatic ring in the present invention, and triazole is also included in this class and classified as an alicyclic ring. The triazole comprises a group generated by the reaction of ethynyl and azido, and also comprises a group generated by the reaction of cycloalkynyl and azido.
Aromatic rings (aromatic rings) include aromatic rings and aromatic heterocycles. Aromatic rings (aryl rings) refer to carbocyclic rings of aromatic hydrocarbon origin, an aromatic all-carbon ring (aromatic all-carbon rings). Aromatic-derived heterocycles (aromatic-derived heterocycles) refer to heterocycles in which a carbon atom of a ring of an aromatic ring is replaced by a heteroatom. For example, aromatic rings include, but are not limited to, benzene, pyridine, pyridazine, pyrimidine, pyrazine, 1,3, 5-triazine, tetrazine (1,2,3,4-, 1,2,4, 5-and 1,2,3, 5-three isomers), indene, indane, indole, isoindole, purine, naphthalene, dihydroanthracene, xanthene, thioxanthene, dihydrophenanthrene, 10, 11-dihydro-5H-dibenzo [ a, d ] cycloheptane, dibenzocycloheptene, 5-dibenzocycloheptenone, quinoline, isoquinoline, fluorene, carbazole, iminodibenzyl, naphthylene, dibenzocyclooctyne, azadibenzocyclooctyne, and the like, substituted versions of any, and hybridized versions of any. Wherein the nitrogen atom of the ring is also allowed to exist in a cationic form. For example, pyridine, pyridazine, pyrimidine, pyrazine are aza forms of benzene, indole, isoindole are aza forms of indene, carbazole is aza form of fluorene, xanthene is oxa form of dihydroanthracene, thioxanthene is thia form of dihydroanthracene, 9H-thioxanthene-10, 10-dioxide is sulfone hybrid form of dihydroanthracene. Pyridinium is a substituted form of pyridine, in which case the nitrogen atom is present in a cationic form. The aromatic ring includes, but is not limited to, paragraphs [267] to [284] in addition to paragraphs [130] to [131] in CN 104530417A. It should be noted that biphenyl in trivalent biphenyl is not a basic cyclic core structure, but is formed by combining a trivalent phenyl cyclic core structure and a divalent phenyl (phenylene); trivalent diphenylmethane is similar to trivalent biphenyl.
Examples of sugar rings (sugar rings) include, but are not limited to, furanose rings, pyranose rings, cyclodextrins, and the like. The sugar ring is selected from the skeleton of cyclic monosaccharide or cyclic monosaccharide derivative
Figure BDA0002963906360000096
Backbones of oligosaccharides or oligosaccharide derivatives
Figure BDA0002963906360000097
Polysaccharide or polysaccharide derivative backbone
Figure BDA0002963906360000101
Any of the above.
Figure BDA0002963906360000102
And preferred versions of the three include, but are not limited to, those described, exemplified and cited in documents CN104530415A, CN104530417A, WO/2016/206540A, CN201610252378X (CN106967213A), CN201710125672.9, CN201710126727.8, CN201911013365.7, CN202010209573.0 and the documents cited therein. Taking CN104530417A as an example, the corresponding segment [0231]～[0234]。
The skeleton of the cyclic monosaccharide or the cyclic monosaccharide derivative has 3, 4, 5, 6 or 7 carbon atoms, and the structure of the skeleton is any one form or a combination form of any two or more forms of an isomer, a chiral isomer, an optical isomer, a conformational isomer and a rotational isomer. Monosaccharides or monosaccharide derivatives having a cyclic monosaccharide backbone of 6 carbon atoms are preferred, and include, by way of example and not limitation, any monosaccharide of glucose, allose, altrose, mannose, gulose, idose, galactose, talose, aloketose, fructose, sorbose, tagatose, inositol. The cyclic structure is preferably a five-membered ring or a six-membered ring.
The skeletons of the oligosaccharides or oligosaccharide derivatives are combined in a cyclic monosaccharide skeleton mode, including but not limited to linear, branched, hyperbranched, dendritic, comb-shaped and cyclic modes. The number of monosaccharide units is 2-10. Taking a cyclic mode as an example, any cyclodextrin of alpha-cyclodextrin, beta-cyclodextrin and gamma-cyclodextrin or derivatives thereof can be formed in a combined mode.
The combination mode of the polysaccharide or polysaccharide derivative skeleton and the cyclic monosaccharide skeleton thereof includes but is not limited to linear, branched, hyperbranched, dendritic, comb-shaped and cyclic modes. The number of monosaccharide units is more than 10. By way of example, D-glucopyranose units are linked in sequence via alpha-1, 4 glycosidic linkages to form linear combinations; the linear structures are connected end to end, so that a ring combination mode can be formed. For another example, when at least one D-glucopyranose unit is bonded to an adjacent glucose unit via at least two glycosidic linkages, such as an alpha-1, 2 glycosidic linkage, an alpha-1, 3 glycosidic linkage, an alpha-1, 4 glycosidic linkage, and an alpha-1, 6 glycosidic linkage, a combination of branching and hyper-branching is formed. When all glucose units are repeatedly linked in a regular manner by specific three or more glycosidic linkages, a comb-like pattern of combinations can be formed. Specifically, the polysaccharide or polysaccharide derivative may be any one of starch, chitin, cellulose, and dextran, for example.
Condensed rings (condensed rings) include, but are not limited to, lactones (e.g., beta-lactide), lactides (cyclic diesters of hydroxycarboxylic acids, e.g., lactide), lactams (e.g., beta-propiolactam), cyclic imides (e.g., maleimide, succinimide, 3H-1,2, 4-triazoline-3, 5-dione), cyclic anhydrides, cyclic peptides, and the like.
The term "substituted" as used herein means that any one or more hydrogen atoms at any position of the "substituted" hydrocarbon group may be substituted, for example, "substituted" or "hydrocarbon group".
Compounds formed by replacement of a carbon atom in at least one position in a hydrocarbon with a heteroatom-containing group are collectively referred to as heterohydrocarbons. The heteroatom in the present invention is not particularly limited, but includes, but is not limited to, O, S, N, P, Si, F, Cl, Br, I, B and the like.
Besides the hydrocarbon group, the substituent group containing hetero atom includes but is not limited to halogenated alkyl, nitro, silicon group (trimethylsilyl, tert-butyldimethylsilyl, trimethoxysilyl, etc.); further, there are included, but not limited to, groups formed by directly connecting a hydrocarbon group or a heterohydrocarbon group to a heteroatom-containing linking group such as an oxy group, a thio group, an acyl group, an acyloxy group, an oxyacyl group, -NH-C (═ O) -, -C (═ O) -NH-, and the like; taking the hydrocarbyl group as an example, a hydrocarbyloxy group, a hydrocarbylthio group, an acyl group, an acyloxy group, a hydrocarbyloxyacyl group, an aminoacyl group, an acylamino group, and the like are formed in this order.
"acyl" includes both carbonyl and non-carbonyl groups, and examples include, but are not limited to, carbonyl, sulfonyl, sulfinyl, phosphoryl, phosphorylidene, nitroxyl, nitrosyl, thiocarbonyl, imidoyl, thiophosphoryl, dithiophosphoryl, trithiophosphoryl, thiophosphorylidene, dithiophosphorylidene, thiophosphoryl, dithiophosphono, thiophosphoryl, and the like. And is preferably carbonyl, thiocarbonyl, sulfonyl or sulfinyl. Unless otherwise specified, acyl refers specifically to carbonyl.
With respect to a compound, a group or an atom, both substituted and hybridized, e.g. by nitrophenyl (nitrophenyl as substituent also containing the heteroatom N), also e.g. -CH 2 -CH 2 -CH 2 -is replaced by-CH 2 -S-CH(CH 3 ) - (both substituted by methyl and hybridized by thio).
"atomic separation" or "atomic distance" refers to the number of atoms of the backbone chain spaced along the backbone chain, regardless of the side groups and side chains, and is typically the shortest atomic distance, and can be used to refer to the length of the linker; for example, in A-CO-NH-B the atomic separation of A and B is 2, A-p-Ph-CH 2 The atomic spacing of A and B in-B is 5(p-Ph is p-phenylene), and as another example A-CH (CH) 2 CH 2 CH 2 CH 3 ) The atomic spacing of B is 1. The "backbone atoms" participating in the formation of the atomic spacer can only be non-hydrogen atoms. Wherein, for divalent linking groups containing cyclic structures, the atomic spacing thereof means the shortest atomic number calculated along the ring-forming atoms, for example, the atomic spacing of p-phenylene, i.e., 1, 4-phenylene, is 4, the atomic spacing of m-phenylene is 3, and the atomic spacing of o-phenylene is 2. And as-CH 2 –、–CH(CH 3 )–、 –C(CH 3 ) 2 –、–CH(CH 2 Ph) 2 –、–C(CH 2 OX) -all had an atomic spacing of 1.
"carbon chain linker" refers to a linker in which all of the main chain atoms are carbon atoms, while the side chain moieties allow for the substitution of a heteroatom or heteroatom-containing group for a hydrogen atom of the main chain carbon. When the "backbone atom" is a heteroatom, it is also referred to as a "backbone heteroatom", e.g., A-S-CH 2 -B、 A-O-CH 2 -B、
Figure BDA0002963906360000111
(the atomic separation is 4) is considered to contain a main chain heteroatom. The carbon chain linking group can be divided into alkylene and a carbon chain linking group of which the side group contains hetero atoms; heteroatom-containing carbon chain linking groups of the pendant groups include, but are not limited to, oxo (═ O), thio (═ S), amino (attached to the backbone carbon by a carbon-nitrogen double bond), oxahydrocarbyl groups in the form of ether linkages, thiahydrocarbyl groups in the form of thioether linkages, azahydrocarbyl groups in the form of tertiary amino groups, and the like.
The "carbon chain linker" backbone is composed entirely of carbon atoms, and the pendant groups of the carbon chain are allowed to contain heteroatoms. I.e. linked by methylene or substituted methylene groups. The substituted methylene group may be substituted with a monovalent substituent, two monovalent substituents or a divalent substituent (e.g. divalent oxygen, e.g. together with the divalent methylene group to form a three-membered ring
Figure BDA0002963906360000112
) And (4) substitution. The substituted methylene group may be one in which a hydrogen atom is substituted (e.g., -CH (CH) 3 ) -, may also be twoEach hydrogen atom being substituted (e.g. - (CH) 3 )C(OCH 3 ) -, it being possible for two hydrogen atoms to be substituted simultaneously (for example carbonyl, thiocarbonyl, -C (-NH) -, -C (-N) + H 2 ) -) or may be cyclic side groups (e.g.
Figure BDA0002963906360000113
The atomic separation is denoted as 1).
Secondary amino or hydrazine bonds in the context of the present invention mean that the "NH-" is capped at both ends by alkylene groups, e.g. -CH 2 -NH-CH 2 -; if-C (═ O) -NH-is referred to as an amide bond, it is not considered to contain a secondary amino bond.
The number of the corresponding repeating units in the "polymer chain" in the present invention is at least 2.
In the present invention, "molecular weight" means the mass size of one molecule of a compound, "average molecular weight" means the mass size of a component of a compound of the formula in a macroscopic material, and "average molecular weight" means generally "number average molecular weight" M "when not specified otherwise n . The number average molecular weight may be the molecular weight of the polydisperse block or substance, or the molecular weight of the monodisperse block or substance. Unless specifically stated otherwise, the units of "molecular weight" and "average molecular weight" are reported in daltons, Da. The molecular weight of the polyethylene glycol chain can also be characterized by the degree of polymerization, specifically the number of repeating units (oxyethylene units, EO units) in one compound molecule. Accordingly, the average, number average, of the number of repeating units is characterized by an "average degree of polymerization", preferably a "number average degree of polymerization".
In the present invention, in the case of polydispersity, "equal" or "the same" or "equal" (including other forms of equivalent expressions) of the molecular weight/degree of polymerization of individual molecules of the compounds, the number average molecular weight/degree of number average polymerization of the components of the compounds in the macroscopic material, are not restricted to strictly equal values in the case of no particular designation, but are close to or approximately equal in index value, which close to or approximately equal preferably deviate by not more than ± 10%, usually based on a predetermined number. "about", "about" and "approximately" generally refer to a range of values of. + -. 10%, and in some cases may be magnified to. + -. 15%, but not more than. + -. 20%. For example, the deviation of 10kDa from 11kDa and 12kDa is 10% and 20%, respectively. As another example, when the molecular weight of a PEG component of a given general formula is 5kDa, the corresponding molecular weight or number average molecular weight is allowed to vary within a range of 5 kDa. + -. 10%, i.e., 4500-5500 Da. In the case of monodispersity, the same or equal numbers of oxyethylene units in the individual compound molecules and in the formula means strictly equal in value; for example, if the number of EO units for a PEG component is set to 11, a value equal to 12 does not fall within the set range; however, in the case of a macroscopic product obtained by a certain preparation method for obtaining a compound component containing a set EO unit number, due to the limitations of the preparation method and the purification method, the macroscopic product may contain impurities of other EO unit number components except for the target EO unit number component, and when the average number of EO units deviates from the preset EO unit number by not more than. + -. 5% (base number. gtoreq.10) or not more than. + -. 0.5 (base number <10), the macroscopic product containing the target component is regarded as being obtained; furthermore, when the content of the component in accordance with the number of EO units or the average number of EO units reaches a certain percentage (preferably ≥ 90%, more preferably > 95%, more preferably > 96%, more preferably > 98%, more preferably 99% to 100%), the macroscopic product obtained falls within the scope of the present invention; even if the above-mentioned content ratio is not attained, the product having an insufficient content, the component appearing as a coproduct or a byproduct, or the like, is obtained within the scope of the present invention regardless of whether separation and purification are carried out, as long as the production method of the present invention or a similar method using substantially the same production concept is employed.
When the molecular weight of the compound formula of the polydisperse composition is described in Da, kDa, number of repeating units, number of EO units, the values fall within a certain range of the given values (including the endpoints, preferably within ± 10%) for a single compound molecule; when the molecular weight of the formula of the compound in which the monodisperse component is described by the oxyethylene unit is predetermined, there is no fluctuation in the range as discrete points, but the product produced may have a fluctuation in the average number of EO units within a certain range (not more than. + -. 10% or. + -. 1, preferably not more than. + -. 5% or. + -. 0.5) due to the nonuniformity of the molecular weight. For example, mPEG has a molecular weight of 5kDa, which means that the molecular weight of a single molecule in the general formula is 4500-5500 Da, the average molecular weight of the corresponding component of the prepared product is 5kDa, that is, the product with the average molecular weight of 4500-5500 Da is the target product, and the component with the molecular weight falling in the range contributes to the content of the target component; also, if mPEG is designed to have 22 oxyethylene units, the number of EO units of all the molecules of the compound in the formula should be strictly 22, but the product may be a mixture of compounds having 20, 21, 22, 23, 24 EO units, where an average number of EO units falling within a range of 22. + -. 2.2 (preferably within a range of 22. + -. 1.1) is considered to be the target component, and components having a molecular weight falling within this range may be considered to be the target component for purity calculation. In the present invention, the product PDI <1.005 is regarded as monodispersity and can be described as PDI ═ 1.
In the invention, the general structural formula of PEG is
Figure BDA0002963906360000121
Wherein one end is connected with L 2 Is connected at the other end with F G And n is any suitable integer (the value corresponding to the average degree of polymerization in the product is not limited to an integer, and may be a non-integer, about n). In the present invention, PEG defines only the general structural formula, and does not limit the molecular weight.
While not limited to the above definitions of molecular weight and number average molecular weight, reference to values in the present disclosure as "about" or "about" generally means a range of values of. + -. 10%, and in some cases may be magnified to. + -. 15% but not more than. + -. 20%. The preset value is used as a base number.
Regarding the polydispersity PDI in the present invention, when other parameters are the same or considered to be the same for different batches of raw materials, as long as the PDI does not exceed a predetermined value, it is considered that there is no significant difference and the raw materials are considered to be the same.
As used herein, a percentage of "about" generally means. + -. 0.5%.
The numerical ranges in the present invention include, but are not limited to, numerical ranges expressed as integers, non-integers, percentages, and fractions, and include both endpoints unless otherwise specified. For example, a value of 4 to 8 includes a left end integer of 4 and a right end integer of 8. As another example, 99% to 100% includes the left endpoint of 99% and the right endpoint of 100%.
"Stable existence" and "degradable" of groups in the present invention are a pair of opposite concepts.
"degradable" (be degradable or can be degraded) means that the cleavage of a chemical bond occurs and that the cleavage is at least two residues independently of one another. If the structure is altered by a chemical change, but the entire linker is still only one complete linker, the linker is still classified as "stably available". The degradable condition is not particularly limited, and may be an in vivo physiological condition, or an in vitro simulated physiological environment or other conditions, preferably an in vivo physiological condition and an in vitro simulated physiological condition. The physiological condition is not particularly limited, and includes, but not limited to, serum, heart, liver, spleen, lung, kidney, skeleton, muscle, fat, brain, lymph node, small intestine, gonad, etc., and may refer to intracellular, extracellular matrix, normal physiological tissue, and pathological physiological tissue (such as tumor, inflammation, etc.). The in vitro simulated environment is not particularly limited and includes, but is not limited to, physiological saline, buffer, culture medium, and the like. The degradation rate is not particularly limited, and may be, for example, rapid degradation by an enzyme, slow hydrolysis under physiological conditions, or the like. The physiological condition in vivo includes physiological condition during treatment, such as ultraviolet irradiation, thermotherapy, etc. Including but not limited to, degradable under conditions of light, heat, low temperature, enzymes, redox, acidic, basic, physiological conditions, in vitro simulated environments, and the like, preferably under conditions of light, heat, enzymes, redox, acidic, basic, and the like. By degradable is meant degradation under stimulation under any of the conditions described above. The light conditions include, but are not limited to, visible light, ultraviolet light, infrared light, near infrared light, mid-infrared light, and the like. The thermal conditions refer to temperature conditions above normal physiological temperature, typically above 37 ℃, and typically below 45 ℃, preferably below 42 ℃. The low temperature condition is lower than the physiological temperature of human body, preferably lower than 25 ℃, more preferably less than or equal to 10 ℃, and specific examples are refrigeration temperature, freezing temperature, liquid nitrogen treatment temperature, 2-10 ℃, 4-8 ℃, 4 ℃, 0 ℃, and-20 +/-5 ℃, and the like. The enzyme conditions are not particularly limited, and enzymes that can be produced under physiological conditions are included, and examples thereof include peptidases, proteases, lyases and the like. The redox conditions are not particularly limited, such as redox transition between a thiol group and a disulfide bond, and hydrogenation reduction transition. The acidic and alkaline conditions mainly refer to the pH conditions of the internal body parts such as normal tissues, diseased tissues, organs or tissues in the treatment period, for example, the stomach is acidic, and the tumor part is often acidic. Degradable herein refers to degradation by metabolic action in vivo (e.g., physiological action, such as enzymes, such as redox, etc.), degradation at specific sites in the body by micro-environmental stimuli (e.g., acidic, basic), or degradation under clinical therapeutic stimuli (e.g., light, such as heat, such as hypothermia), etc. It should be noted that some extreme conditions in organic chemistry relative to organisms, such as bond cleavage under strong acid, strong base, high temperature (e.g., above 100 ℃), etc., are not included in the scope of the degradable conditions of the present invention. For another example, although ether linkages can be cleaved under strong acid conditions such as hydrobromic acid, they are always classified as stably available linkers in the present invention.
In contrast, a linker is defined as "stably present" (be table or can remaining table) as long as it remains as an intact linker (one linker is stably covalently linked to its neighboring groups), wherein chemical changes that preserve the integrity of the linker are allowed to occur. The chemical changes are not particularly limited and include, but are not limited to, isomerization, oxidation, reduction, ionization, protonation, deprotonation, substitution reactions, and the like. The conditions that can be stably present are not particularly limited, and include, but are not limited to, light, heat, low temperature, enzymes, redox, neutrality, acidity, alkalinity, physiological conditions, in vitro simulated environment, and the like, and preferably, light, heat, enzymes, redox, acidity, alkalinity, and the like. The stable existence here means that stable linkage can be maintained in the metabolic cycle in vivo without specific stimulation (such as pH condition at a specific site, light, heat, low temperature at the time of treatment, etc.) and the decrease in molecular weight due to chain cleavage (as long as integrity is maintained) does not occur.
In addition, the term "stably exist" is not an absolute concept for the same linker, for example, an amide bond is more stable under acidic or basic conditions than an ester bond, and the linker "stably exist" in the present invention includes an amide bond. However, for example, a peptide bond, an amide bond formed by the dehydrocondensation of an α -carboxyl group of one molecule of an amino acid and an α -amino group of one molecule of an amino acid, can be cleaved when subjected to a particular enzyme, and is therefore also included in a "degradable" linker. Similarly, carbamate, thiocarbamate, or the like may be a stably existing linker or a degradable linker. More generally, carbamate groups, thiocarbamate groups, and the like are more prone to slow degradation, while amide bonds other than peptide bonds may be stable during in vivo circulation. Also, for example, common ester bonds can be degraded under acid and alkali conditions, while ester bonds contained in a special structure can be degraded under ultraviolet light conditions. For another example, even if some chemical bonds are degraded by a specific enzyme, if they are used clinically, the corresponding chemical bonds can be considered to be stably present if the circulating pathway does not pass through or substantially passes through the specific enzyme environment (such as in the case of site-specific administration).
In order to define the degradable nature of the compound structure more clearly, a reference criterion is provided, i.e. the chemical bonds under investigation are kept within a certain percentage (e.g. 90%) for a limited time interval. In the case of 90%, the pharmacokinetic profile of the functionalized polyethylene glycol modified product is usually used as a reference, based on the percentage of the dose that meets the clinical evaluation criteria. For example, for an intravenously administered pegylated drug, when the blood concentration (based on the active drug ingredient, including the pegylated drug and the non-pegylated ingredient after degradation) is less than 15% of the initial concentration (or other ratio more consistent with clinical evaluation of the drug), based on the remaining 85%, it is within the scope of the invention that it is a group that can exist stably if the ratio of one linker group remaining chemically bonded exceeds 90%, and conversely it is a degradable group if it is less than 90%.
Hydrolytic stabilization, enzymatic degradation, and the like reported in the published literature are also incorporated into the present invention. Taking the hydrolysis stability as an example, the hydrolysis rate when the hydrolysis is stable as reported in the published literature is included, preferably the hydrolysis rate under physiological conditions is less than 1-2% (generally 2%), by mass or molar amount per day. The rate of hydrolysis of typical chemical bonds can be found in most standard chemical manuals.
For pegylated drugs that circulate in the blood, the ether linkage CH between the repeating units of the polyethylene glycol component 2 CH 2 -O-CH 2 CH 2 Are generally considered stable to the art. A reference may be provided for distinguishing "stably present" from "degradable", but this is not a critical criterion, i.e. a comparison reference for "off or on" (on/off). A linker is considered to be stably present when it is more stable in the environment than the ether linkage in the PEG fragment.
Similar to the definition of "multivalent" in the valence state, the same number of functional groups is present for the polyfunctional compound. The "functional group" in the present invention means a functional group, and is preferably a reactive group, a protected reactive group, a precursor of a reactive group, or the like. "polyol" means that the number of functional groups is at least 3, e.g., polyol means a compound having at least 3 hydroxyl groups, polythiol means a compound having at least 3 mercapto groups, and the like. It is to be noted that other types of heterogeneous functional groups are allowed, for example, tris (hydroxymethyl) aminomethane is a triol having one amino group, and citric acid is a tricarboxylic acid having one hydroxyl group.
"heterogeneous functional group" in the present invention means another functional group different from a certain type of functional group, for example, in the case of a triol, in which an amino group is a heterogeneous functional group with respect to a hydroxyl group. The "pair of heterofunctional groups" in the present invention are mutually heterogeneous functional groups. "heterogeneous functional groups" and "heterofunctional group pairs" are not limited to describing different functional groups present in the same molecule, but can also describe different functional groups present in different molecules or structures, such as reacting an alkynyl-containing compound with an azido-containing compound, the alkynyl and azido groups being heterogeneous with respect to each other, and also a set of "heterofunctional group pairs".
The term "functional group source" as used herein refers to a source that is reactive or potentially reactive, photosensitive or potentially photosensitive, targeted, or potentially targeted. The term "latent" refers to a chemical moiety that can be converted into a reactive group by a chemical process selected from the group consisting of, but not limited to, functional modification (e.g., grafting, substitution, etc.), deprotection, salt complexation and decomplexing, ionization, protonation, deprotonation, change in the dissociated group, etc., and that can emit light or generate targeting by external stimuli such as light, heat, enzymes, specific binding molecules, in vivo microenvironment, etc. The luminescence is not particularly limited, and includes, but is not limited to, visible light, fluorescence, phosphorescence, and the like.
The change form in the invention refers to a structure form which can be converted into a target reactive group through any chemical change process of oxidation, reduction, hydration, dehydration, electronic rearrangement, structural rearrangement, salt complexation and decomplexing, ionization, protonation, deprotonation, substitution, deprotection, leaving group change and the like.
"reactive group modification" as used herein refers to a modification in which a reactive group remains active (i.e., remains a reactive group) after undergoing at least one chemical change, such as oxidation, reduction, hydration, dehydration, electronic rearrangement, structural rearrangement, salt complexation and decomplexing, ionization, protonation, deprotonation, substitution, deprotection, or change in the form of the dissociated group, or a modification in which a reactive group remains inactive after being protected.
The term "micro-modification" as used herein refers to a chemical modification process that can be accomplished by a simple chemical reaction process. The simple chemical reaction process mainly refers to chemical reaction processes of deprotection, salt complexation and decomplexing, ionization, protonation, deprotonation, leaving group conversion and the like.
"minor variations" correspond to "minor modifications" and refer to structural forms that can form the target reactive group after undergoing simple chemical reaction processes such as deprotection, salt complexation and decomplexing, ionization, protonation, deprotonation, leaving group conversion, and the like. The conversion of the leaving group, such as the conversion of the ester form to the acid chloride form.
"any suitable" in "any suitable linking group", "any suitable reactive group", and the like means a structure that conforms to the basic principle of chemical structure and enables the production method of the present invention to be favorably carried out. The chemical structures described in this way can be regarded as having a clear, defined range.
When at least two structure types are recited, "any combination" of the recited structure types refers to a combination of any two or more of the aforementioned recited related structure types; the number of the structural units is not limited, the number of any structural unit can be zero, one or more than one, when the number of the structural units of the same type is more than 1, the structural units of the same or different chemical structures can be formed, and the sum of the number of the structural units is at least 2. For example, any combination of alkylene, divalent cycloalkyl, divalent cycloalkenyl, divalent cycloalkynyl, divalent cycloalkadienyl, arylene, carbon-carbon double bond, carbon-carbon triple bond, conjugated carbon-carbon double bond, divalent lipoheterocyclic linkage, divalent heteroaromaticheterocyclic linkage, pendant heteroatom-containing carbon chain linkage is exemplified by-Ph-CH 2 -Ph- (arylene-alkylene-arylene), -CH 2 -Ph-CH 2 CH 2 - (alkylene-arylene-alkylene, wherein the number of alkylene groups is 2 and has a different chemical structure), or the structure of the aforementioned examples in which the benzene ring is replaced by hexylene, diazacyclo, 1- (2-pyridyl) hexahydro-1H-1, 4-diazepin. As another example, cycloalkenylhydrocarbyl + hydrocarbylene as substituents for the hydrocarbyl group, and cycloalkadienylhydrocarbyl as substituents for the hydrocarbyl group. The alkylene group, that is, the divalent alkyl group in the present invention includes an open chain alkylene group which means a divalent alkyl group not containing a cyclic structure and a divalent cycloalkyl group which means a divalent alkyl group containing a cyclic structure.
"alternative ranges including, but not limited to," means that structures within the stated ranges are optional and not limited to structures within the stated ranges, but not all structures within the stated ranges are applicable, especially if the invention specifically excludes them from candidate lists. The basic principle is that the preparation method of the invention is successfully implemented as a screening standard.
"aminocarboxylic acid" in the present invention means an NH group at one end 2 And compounds with a COOH terminal, in addition to various natural amino acids, also include some non-natural compounds. Omega-amino carboxylic acids are also preferred. The omega-aminocarboxylic acids of the invention are preferably NH 2 -L 5 -COOH, wherein L 5 Is any one of alkylene, arylene or any combination thereof, and contains at least two carbon atoms, such as methylene, ethylene (1, 2-ethylene or 1, 2-ethylene-CH (CH) 3 ) -), propylene, 1, 4-cyclohexylene, phenylene, benzylidene, -CH (Bn) -, where Bn is benzyl. Wherein arylene means that both covalent bonds are directly from an aromatic ring. The omega-aminocarboxylic acid is preferably an omega-aminoalkanecarboxylic acid, in which case L 5 Is alkylene (which may be open-chain or cyclic); further preferred is H 2 N(CH 2 ) j1 COOH, wherein, the integer j 1 The amino acid is selected from 2 to 20, preferably 2 to 12, more preferably 2 to 6, and examples thereof include 3-aminopropionic acid (. beta. -alanine), 4-aminobutyric acid (. gamma. -butyrate), 5-aminopentanoic acid, 6-aminocaproic acid, 7-aminoheptanoic acid, and 8-aminocaprylic acid.
The source of the amino acid in the present invention is not particularly limited unless otherwise specified, and may be a natural source, a non-natural source, or a mixture of both. The amino acid structure type in the present invention is not particularly limited unless otherwise specified, and may be any type as long as it is L Type-can also mean D Type-or a mixture of both.
The definitions and examples of the amino acid skeleton, the amino acid derivative skeleton and the cyclic monosaccharide skeleton in references CN104877127A, WO/2016/206540A, CN201610252378X (CN106967213A), CN201710125672.9 and CN201710126727.8 and in the respective citations are also incorporated by reference into the present invention. The amino acid skeleton refers to a residue having basic characteristics of an amino acid, and specifically refers to a residue formed by losing a carboxyl hydroxyl group (including all C-terminal carboxyl hydroxyl groups, and also including carboxyl hydroxyl groups on side groups such as aspartic acid and glutamic acid), a hydrogen atom on a hydroxyl group, a hydrogen atom on a phenolic hydroxyl group (tyrosine), a hydrogen atom on a mercapto group (such as cysteine), a hydrogen atom on a nitrogen atom (including all N-terminal hydrogen atoms, and also including hydrogen atoms in amino groups in side groups such as lysine, a hydrogen atom on an epsilon-amino group on ornithine, a hydrogen atom in an amino group on a ring of histidine and tryptophan, and the like), an amino group on an amide (such as asparagine, glutamine, and the like), an amino group in a guanidine group, or a hydrogen atom in an amino group. The amino acid derivative skeleton means an atom or group portion having its essential characteristics in addition to the amino acid skeleton.
In the present invention, the "C-carboxyl group" and "N-amino group" of an amino acid are both alpha-positions unless otherwise specified.
"biologically relevant substances" include, but are not limited to, the substances described, exemplified and cited in documents CN104877127A, WO/2016/206540A, CN201610252378X (CN106967213A), CN201710125672.9, CN201710126727.8 and the respective cited documents. In general terms, biologically relevant substances include, but are not limited to, the following: drugs, proteins, polypeptides, oligopeptides, protein mimetics, fragments and analogs, enzymes, antigens, antibodies and fragments thereof, receptors, small molecule drugs, nucleosides, nucleotides, oligonucleotides, antisense oligonucleotides, polynucleotides, nucleic acids, aptamers, polysaccharides, proteoglycans, glycoproteins, steroids, lipid compounds, hormones, vitamins, phospholipids, glycolipids, dyes, fluorescent substances, targeting factors, targeting molecules, cytokines, neurotransmitters, extracellular matrix substances, plant or animal extracts, viruses, vaccines, cells, vesicles, liposomes, micelles, and the like. The biologically-relevant substance may be a biologically-relevant substance itself, or a precursor, an activated state, a derivative, an isomer, a mutant, an analog, a mimetic, a polymorph, a pharmaceutically-acceptable salt, a fusion protein, a chemically-modified substance, a gene recombinant substance, or the like thereof, or a corresponding agonist, an activator, an inhibitor, an antagonist, a modulator, a receptor, a ligand or a ligand, an antibody or a fragment thereof, an acting enzyme (e.g., a kinase, a hydrolase, a lyase, an oxidoreductase, an isomerase, a transferase, a deaminase, a deiminase, a invertase, a synthetase, or the like), a substrate for an enzyme (e.g., a substrate for a coagulation cascade protease, or the like), or the like. The derivatives include, but are not limited to, glycosides, nucleosides, amino acids, and polypeptide derivatives. The chemical modification products formed by modifying the reactive groups to change types, additionally introducing structures such as functional groups, reactive groups, amino acids or amino acid derivatives, polypeptides and the like are chemical modification substances of biologically related substances. The bio-related substance may also allow for a target molecule, adjunct or delivery vehicle to be bound thereto, either before or after binding to the functionalized polyethylene glycol, to form a modified bio-related substance or a complexed bio-related substance. Wherein, the pharmaceutically acceptable salt can be inorganic salt, such as hydrochloride, sulfate and phosphate, and can also be organic salt, such as oxalate, malate, citrate, etc. The term "drug" as used herein includes any agent, compound, composition or mixture that provides a physiological or pharmacological effect, either in vivo or in vitro, and often provides a beneficial effect. The class is not particularly limited and includes, but is not limited to, pharmaceuticals, vaccines, antibodies, vitamins, foods, food additives, nutritional agents, nutraceuticals, and other agents that provide a beneficial effect. The "drug" is not particularly limited in the range that produces physiological or pharmacological effects in vivo, and may be a systemic effect or a local effect. The activity of the "drug" is not particularly limited, and is mainly an active substance that can interact with other substances, and may also be an inert substance that does not interact with other substances; however, inert drugs can be converted to the active form by in vivo action or some stimulus. Wherein, the small molecule drug is a biological related substance with the molecular weight not more than 1000Da, or a small molecule mimicry or an active fragment of any biological related substance.
When the number of a certain symbol in a molecule of the present invention is 2 or more than 2Without being specifically stated, have the same structure or polymer formula, for which different molecular weights are allowed. Such as Q, Q in the invention 3 、Q 5 、 Q e The same symbols define that when the number is greater than 1 in the same molecule, any two may have different structures.
The "electron-altering group" in the present invention means that the electron cloud density on the unsaturated structure (particularly, aromatic ring structure) can be altered with respect to the hydrogen atom. Taking an aromatic ring as an example, comprehensively considering the sum of the induction effect, the conjugation effect and the super-conjugation effect of the electron change group on the aromatic ring, the electron cloud density of the aromatic ring is increased to be an electron-donating group, and the electron cloud density of the aromatic ring is decreased to be an electron-withdrawing group. Examples of the electron-donating group include a lower alkoxy group, a lower alkylamino group, a di-lower alkylamino group, a lower alkyl group, an aryloxy group, an aralkyloxy group, an aminoaryl group, a hydroxyl group, an amino group, a mercapto group, an alkylthio group and the like. A typical example of a lower alkyl group referred to in the present invention is C 1-6 Alkyl, including but not limited to lower alkoxy, lower alkylamino, di-lower alkylamino, lower alkyl of lower alkyl. Examples of the electron-withdrawing group include a halogen group (bromine group, chlorine group, fluorine group, iodine group and the like), a nitro group, a trihalomethyl group, a cyano group, a carboxyl group, a formyl group, a keto group, an azo group, an amidocarbonyl group, an amidosulfonyl group, a carboxamide group, a sulfoxy group, a sulfonamide, a urea group and the like. In addition, such as benzene rings, double bonds, either as electron donating groups or electron withdrawing groups, depend on the overall effect on the electron cloud density.
In the present invention, for a single bond derived from a cyclic structure, when it is not labeled to a specific ring-forming atom, but is directed to the inside of the ring through a covalent bond between two ring-forming atoms, e.g.
Figure BDA0002963906360000161
It means that the single bond may be located at any suitable position on the ring; wherein the symbol of the atom directed to the end inside the ring represents a ring-forming atom (e.g., M), and the symbol directed to the end outside the ring represents a hydrogen atom or a substituent (e.g., Q, Q) 5 、Q e ) (ii) a Inside of the directional ringAnd no other label, represents a ring-forming atom attached at any suitable position, points outward and terminates with a wavy line to represent a free base end, which may form a covalent single bond with other groups.
1. A nitrogen-branched nine-arm polyethylene glycol derivative has a structure shown in a general formula (1):
Figure BDA0002963906360000162
wherein N is a nitrogen atom trivalent branching center;
U 0 is a tetravalent radical, 3U's in the same molecule 0 The same; a nitrogen atom with a trivalent branching center and 3U 0 Together form a nine-valent branched structure;
L 2 is absent or L 2 Is a divalent linking group; in the same molecule, 9L 2 Are the same or different from each other;
PEG is a polyethylene glycol block taking ethylene oxide as a repeating unit, and both ends of the PEG are oxygen groups;
F G is a hydrogen atom or a terminal functional group; when F is present G Containing at least one terminal functional group R when not a hydrogen atom 01 (ii) a Wherein R is 01 Is a functional group capable of interacting with biologically-relevant substances.
Preferably, when F G When it is a terminal functional group, F G Is- (L) 0 -G) g -(F) k Wherein g is 0 or 1; g is a terminal branching group selected from a trivalent or higher valent connecting group which connects the PEG chain segment with a terminal functional group; l is 0 Is a divalent linking group which connects the PEG chain segment with the terminal branching group G; the integer k is the number of F in a single functionalized end and is selected from 1 or 2-250; f contains functional group, and the structure of F is- (Z) 2 ) q -(Z 1 ) q1 -R 01 Wherein q and q1 are each independently 0 or 1, Z 1 、Z 2 Each independently is a divalent linking group, R 01 Is a functional group capable of interacting with a biologically relevant substance; f is hydrogen atomAnd form a terminal functional group of hydroxyl, amino or sulfhydryl;
when g is 0, k is 1, L 0 G is not present, F is not a hydrogen atom, hydroxyethyl, hydroxyl terminated PEG chain;
when G is 1, G is present, L 0 And k may be present or absent, and k is 2 to 250, where F is allowed to be a hydrogen atom.
1.1. Trivalent branching center of nitrogen atom
The nitrogen atom trivalent branching center is derived from a trifunctional small molecule, any of whose residues can be trivalent nitrogen atoms, preferably from any of the following trifunctional small molecules:
Figure BDA0002963906360000171
1.2. Tetravalent radical U 0
U 0 Comprises
Figure BDA0002963906360000172
Any of the tetravalent core structures; wherein the star ends in the quaternary structure point to the trivalent branching center of the nitrogen atom and the three non-star ends point to the three divalent linking groups L 2 ；
The U is 0 Further preferably, the compound has any one of the following structures:
Figure BDA0002963906360000173
Figure BDA0002963906360000181
Figure BDA0002963906360000182
wherein j is 1 Is 0 or 1, n i1 Is 0, 1 or 2, n i2 Is 0, 1, 2 or 3;
the U is 0 Further selected from any one of the following structures:
Figure BDA0002963906360000191
Figure BDA0002963906360000201
Figure BDA0002963906360000211
Figure BDA0002963906360000221
wherein the asterisk end of the quadrivalent structure points to the trivalent branching center of the nitrogen atom, and the three non-asterisk ends point to three divalent linking groups L 2 。
1.3. Divalent linking group L U0
Nitrogen atom trivalent branching center and U 0 Covalently linked moieties excluding nitrogen and U 0 The part of the branched core of (A) is denoted by L U0 ， L U0 Y is preferably a divalent linking group containing an ether bond (-O-), a thioether bond (-S-), an ester bond (-C (O) O-, or-OC (O) -), an amide bond (-C (O) NH-, or-NHC (O) -), a carbonate bond (-OC (O) O-), a carbamate bond (-NHC (O) O-, or-OC (O) NH-), or a urea bond (-NHCONH-). "trivalent branching center of nitrogen atom and U 0 The atomic distance between the branched cores is more than or equal to 1, preferably 1 to 100, namely, the atomic distance can be selected from any one of 1, 2, 3, 4, 5, … …, 98, 99 and 100, or from any one of 1 or 1 to 10, 10 to 20, 20 to 50 and 50 to 100, and the interval does not include a left end point and includes a right end point.
Said "U" is 0 By branched core "is meant a tetravalent radical U 0 The simplest tetravalent portion of (a), excluding the divalent linking group with the nitrogen atom branching center; 'U' is provided 0 Any of the base atoms at the free ends of the "branched cores" of (A) is indispensable for constituting tetravalent phases, and together constitute tetravalent U 0 The outermost periphery of the nucleus skeleton of (A) lacks any one of the free-base terminalsAnd all of them result in reduction of valence or incomplete structure.
The "nitrogen atom trivalent branching center and U 0 By atomic distance between the branched nuclei of "is meant the distance U from the nitrogen atom 0 The branching nuclei of (a) are separated by atoms.
“U 0 Judgment of branched core of (1) "exemplifies: u in example 5 0
Figure BDA0002963906360000222
U in example 6 0
Figure BDA0002963906360000223
U in example 7 0
Figure BDA0002963906360000224
Etc. are all branched nuclei>C<(ii) a U in example 8 0
Figure BDA0002963906360000225
The branched core of (A) is
Figure BDA0002963906360000226
Example 9U 0
Figure BDA0002963906360000227
The branched core of (A) is
Figure BDA0002963906360000228
And so on.
L U0 When U is taken as an example 0
Figure BDA0002963906360000229
When, L U0 is-CH 2 CH 2 OCH 2 -, containing ether linkages, trivalent branching centres for nitrogen atoms and U 0 Has an atomic distance of 4; when U is turned 0 Is composed of
Figure BDA00029639063600002210
When L is U0 is-CH 2 C (O) NH-containing an amide bond, a trivalent branching center for the nitrogen atom and U 0 The atomic distance between the branched nuclei of (a) is 3.
1.4. Divalent linking group L 2 、L 0 (g＝1)、Z 1 、Z 2
L 2 Is absent, or L 2 To be bound to a tetravalent radical U 0 And a divalent linking group of PEG segment, 9L 2 Are all the same.
L 2 Are groups that can be stably present or degradable. When L is 2 In the case of a group which can be present stably, L 2 Preferred are alkyl groups including, but not limited to, methyl, ethyl, n-propyl, isopropyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, eicosyl, benzyl, substituted C 1-20 Alkyl, substituted aryl, substituted C 1-20 An open-chain heterocarbon group, a substituted heteroaromatic hydrocarbon group, or the like. Wherein, butyl includes but is not limited to n-butyl and tert-butyl. Octyl includes, but is not limited to, n-octyl, 2-ethylhexyl. Wherein the substituent atom and the substituent are selected from any one of a halogen atom, a hydrocarbon substituent and a hetero atom-containing substituent, and preferably a fluorine atom, a chlorine atom, a bromine atom, an iodine atom, C 1-6 Alkyl, alkoxy or nitro.
L in the above general formula (1) 0 When present, L is not a hydrogen atom 2 、L 0 (g＝1)、Z 1 、Z 2 Are each a divalent linking group, and each is independent of the other, and may be the same as or different from each other in the same molecule.
L 2 、L 0 (g＝1)、Z 1 、Z 2 The structure of (a) is not particularly limited, and each independently includes, but is not limited to, a linear structure, a branched structure, or a cyclic-containing structure.
L 2 、L 0 (g＝1)、Z 1 、Z 2 The number of the non-hydrogen atoms of (b) is not particularly limited, and each is independently preferably 1 to 50 non-hydrogen atoms; more preferably 1 to 20 non-hydrogen atoms; furthermore, the utility modelPreferably 1 to 10 non-hydrogen atoms. The non-hydrogen atom is a carbon atom or a heteroatom. The heteroatoms include, but are not limited to, O, S, N, P, Si, B, and the like. When the number of non-hydrogen atoms is 1, the non-hydrogen atoms may be carbon atoms or hetero atoms. When the number of non-hydrogen atoms is more than 1, the kind of non-hydrogen atoms is not particularly limited; may be 1 species, or may be 2 or more than 2 species; when the number of non-hydrogen atoms is more than 1, any combination of carbon atoms and carbon atoms, carbon atoms and hetero atoms, and hetero atoms may be used.
L 2 、L 0 (g＝1)、Z 1 、Z 2 Each independently preferably having 1 to 50 non-hydrogen atoms; wherein the non-hydrogen atom is C, O, S, N, P, Si or B; when the number of the non-hydrogen atoms is more than 1, the kind of the non-hydrogen atoms is 1, 2, or more than 2, and the non-hydrogen atoms are any combination of carbon atoms and carbon atoms, carbon atoms and heteroatoms, and heteroatoms.
L 2 、L 0 (g＝1)、(Z 2 ) q -(Z 1 ) q1 Is not particularly limited, and any one of the divalent linking groups or any one of the divalent linking groups consisting of a group adjacent to a heteroatom is independently a stably existing linking group STAG or a degradable linking group DEGG. For the preferred case, L 0 (g＝1)、(Z 2 ) q -(Z 1 ) q1 Any one of the divalent linking groups or any one of the divalent linking groups consisting of a group with an adjacent heteroatom is independently a stably available linking group STAG or a degradable linking group DEGG.
1.5. Degree of polymerization and dispersibility of polyethylene glycol chain
In the general formula (1), PEG has the general formula
Figure BDA0002963906360000231
Wherein n is the polymerization degree of a polyethylene glycol chain and is selected from 1-2000; the polymerization degrees of the nine PEG chains may be the same as or different from each other, and are respectively represented by n 1 、n 2 、n 3 、n 4 、n 5 、n 6 、n 7 、n 8 、n 9 . Both to allow the same divisionThe numbers of EO units of the nine PEG chains in a subgroup are the same or different from each other, and it is also permissible that the degrees of polymerization of the nine PEG chains in the macropolymer are the same or different from each other. The nitrogen-branched nine-arm polyethylene glycol can be polydisperse or monodisperse. Wherein, the PEG chain n i (i ═ 1,2,3,4,5,6,7,8,9) can be both monodisperse, both polydispersions, or any combination of monodisperse and polydispersions, preferably both monodisperse and polydispersions.
Nine polyethylene glycol chains are obtained in a polymerization mode, and the polydispersity is the same.
The nitrogen-branched nine-arm polyethylene glycol, PDI, obtained by the coupling method depends on the polydispersity property of the raw material, and preferably nine polyethylene glycol chains are both monodisperse or both polydisperse.
In the present invention, the "molecular weight" is not particularly limited, and is "number average molecular weight", and M n It may be either a polydisperse block or a molecular weight of a material or a monodisperse block or a molecular weight of a material, unless otherwise specified, generally referring to polydisperse polymers. When not specifically written, the units are daltons, Da.
PEG chain n for polydispersity i (i ═ 1,2,3,4,5,6,7,8or 9), the number average degree of polymerization of which is preferably from 2 to about 1500; more preferably from 2 to about 1000; more preferably from 2 to about 500; more preferably from 5 to about 500; more preferably from about 11 to about 500; more preferably from about 22 to about 500; more preferably from about 30 to about 250; more preferably from about 34 to about 150. The more preferred the above, the more conventional the molecular weight of the corresponding PEG chain segment is, the simpler and easier the preparation, and the narrower the PDI (polydispersity index) of the molecular weight, the more uniform the performance. The number average molecular weight of linear PEG obtained by the conventional polymerization method is about 2kDa to 40kDa, and in the present invention, the number average molecular weight of each PEG of the nitrogen-branched nine-arm polyethylene glycol derivative also prepared by the polymerization method is selected from 2kDa to 40 kDa. In the context of the present invention, the number-average molecular weight is also preferably about 500,600,700,800,900,1000,1500,2000,2500,3000,3350,3500,4000,5000, 5500,6000,6500,7000,7500,8000,8500,9000,9500,10000,11000,12000,13000,14000,15000, 16000,17000,18000,19000 or 20000, in Da. More preferably, about 1000,1500, 2000,2500,3000,3350, 3500,4000,5000,5500,6000,6500,7000,7500,8000,8500,9000,9500,10000,11000 or 12000 Da. More preferably about 1000,1500,2000,3000,3350,3500,4000,5000,6000,7000,8000,9000 or 10000 Da. More preferably about 1000,1500,2000,3350,3500,4000,5000 or 6000 Da.
For a monodisperse PEG block, the molecular weight is defined by the number of oxyethylene units (reported as EO units). The number of EO units of monodisperse polyethylene glycols prepared according to the prior art is between about 1 and 70, including but not limited to the EO units recited in references { Extert Rev.mol.Diagn.2013,13(4), 315-. Typical numbers of EO units for monodisperse PEG include, but are not limited to, 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 12, 13, 16, 20, 22, 24, 27, 29, 36, 44, 48, 56, 62, 64, 67, and the like. It is specifically noted that the polydispersity of the nitrogen-branched nine-arm polyethylene glycol of formula (1) is determined collectively by the combination of nine PEG chains, and that the nitrogen-branched nine-arm polyethylene glycol species may be a single component or a mixture of different components, as long as the polymer has a PDI of 1. When a single component, nine PEG chains have the same number of EO units. When a mixture of different components, the total molecular weight of each nitrogen-branched nine-arm polyethylene glycol molecule in the polymer is fixed, but the number of EO units in the nine PEG chains therein may each independently be the same or different. Preferably the relative mole percentages of PEG chain components of different numbers of EO units are determined. Most preferably nine PEG chains have the same number of EO units. When a mixture of different components is used, the corresponding number average degree of polymerization may be an integer or a non-integer. Polymers composed of monodisperse blocks of different EO unit numbers still form polydisperse blocks or species if the contents of the individual components are undefined and the PDI is greater than 1. The number of EO units of the monodisperse PEG block is preferably 2-70; more preferably 3 to 70; more preferably 3 to 50; more preferably 3 to 25. The more preferable, the more various the production method. The number of EO units of the monodisperse PEG chain is preferably selected from any of 2, 3, 4, 5, 6,7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 18, 20, 22, 24, 26, 27, 28, 29, 30, 32, 34, 36, 38, 40, 42, 44, 45, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 67, 68, 70.
For the entire nitrogen-branched nine-arm polyethylene glycol derivative, the polydispersity coefficient may be the same or different from that of the individual PEG chains. But the lower the PDI of the whole compound, the better. Therefore, for the chain length distribution of nine PEG chains of the nitrogen-branched nine-arm polyethylene glycol derivative represented by the general formula (1), n is preferable 1 ≈n 2 ≈n 3 ≈n 4 ≈n 5 ≈n 6 ≈n 7 ≈n 8 ≈n 9 (in which case the number average molecular weights of the nine chains may each independently be the same or close to each other) or n 1 ＝n 2 ＝n 3 ＝n 4 ＝n 5 ＝n 6 ＝n 7 ＝n 8 ＝n 9 (nine chains in this case have a fixed molecular weight and are equal to each other). The PEG chains have equal or similar chain lengths, so that the biological related substances modified by the PEG chains can obtain a uniform structure more easily, and the purity and the performance of the product are improved. n is 1 ≈n 2 ≈n 3 ≈n 4 ≈n 5 ≈n 6 ≈n 7 ≈n 8 ≈n 9 The situation applies to polydisperse structures, which can satisfy the requirements of different molecular weights, and n 1 ＝n 2 ＝n 3 ＝n 4 ＝n 5 ＝n 6 ＝n 7 ＝n 8 ＝n 9 The situation of (2) is suitable for a monodispersity structure, the product structure is controlled more accurately, and a modified product with better quality can be obtained.
1.6. Functional group (F)
The functional group F is- (Z) 2 ) q -(Z 1 ) q1 -R 01 ，-(Z 2 ) q -(Z 1 ) q1 -R 01 See document CN104877127A mid-section [0280 ]]～[0505]The description of (A) is not repeated herein. The following description is made for the case where F is not a hydrogen atom, in which case the functional group R is contained 01 。
1.6.1. Functional group R 01 Definition of (1)
R 01 Is a functional group capable of interacting with biologically relevant substances. The interaction with the bio-related substance includes, but is not limited to, formation of covalent bonds, formation of hydrogen bonds, fluorescence, and targeting. R 01 Selected from reactive groups, variations of reactive groups, functional groups with therapeutic targeting properties, functional groups with fluorescent properties. The reactive group is reactive, forms a linkage by a bonding reaction with a biologically relevant substance, and mainly means a reaction of forming a covalent bond, and when forming a non-covalent linkage, performs a complex through a double hydrogen bond or multiple hydrogen bonds. The covalent bond includes, but is not limited to, a covalent bond that can exist stably, a degradable covalent bond, and a dynamic covalent bond. Such variations include, but are not limited to, precursors to reactive groups, reactive forms that are precursors thereto, substituted reactive forms, protected forms, deprotected forms, and the like. The precursor of the reactive group refers to a structure which can be converted into the reactive group through at least one process of peroxidation, reduction, hydration, dehydration, electronic rearrangement, structural rearrangement, salt complexation and decomplexing, ionization, protonation, deprotonation and the like. The precursor may be reactive or non-reactive. The variation of the reactive group refers to a reactive group which is still active (still reactive group) after at least one process of oxidation, reduction, hydration, dehydration, electronic rearrangement, structural rearrangement, salt complexation and decomplexing, ionization, protonation, deprotonation, substitution, deprotection, etc., or an inactive form after being protected. Fluorescent functional groups can be classified as fluorescent functional groups as long as the fluorescent functional groups can emit fluorescence, or can emit fluorescence through in vivo microenvironment action (such as fluorescein diacetate) or can emit fluorescence through clinical stimulation (such as light stimulation, thermal stimulation and the like). The dynamic covalent bonds include, but are not limited to { Top Curr Chem (2012)322: 1-32}, { Top Curr Chem (2012)322:291-, { Synlett 2016; 177-.
R 01 Including but not limited to groups A through H or variations thereof, groups R 01 Is a reactive group or a modified form thereof is a reactive group.
Class A: active ester groups (including, but not limited to, succinimide active ester groups (e.g., A1, A6), p-nitrophenyl active ester groups (e.g., A2, A7), o-nitrophenyl active ester groups (e.g., A11, A12), benzotriazole active ester groups (e.g., A5, A10), 1,3, 5-trichlorobenzene active ester groups (e.g., A3, A8), fluorophenyl active ester groups (e.g., A13, e.g., 1,3, 5-trifluorobenzene active ester groups, pentafluorobenzene active ester groups), imidazole active ester groups (e.g., A4, A9)), and similar structures of active ester groups A16-A18 (e.g., 2-thione-3-thiazolidinecarboxylate (tetrahydrothiazole-2-thione-N-formate), 2-sulfoxothiazolidine-3-carboxylate, 2-pyrrolidine-N-carboxylate, 2-thione-N-formate, succinimide-formate, and p-nitrophenyl-butyrophenolate groups (e.g., a-butyrophenolate, a-tetrazolium, and a mixture thereof), and a salt thereof, 2-thioketone benzothiazole-N-formate group, 1-oxo-3-thiooxoisoindoline-N-formate group, etc.);
Class B: sulfonate, sulfinate, sulfone, sulfoxide, 1, 3-disulfonyl-2-propylcarbonylphenyl, sulfone methacryl, and the like;
class C: hydroxylamino group, mercapto group, amino group (primary amino group such as C4, or secondary amino group such as C5, C15), halogen atom, haloacetamido group (such as iodoacetamido group), tetramethylpiperidinyloxy group, dioxopiperidinyloxy group (3, 5-dioxo-1-cyclohexylamine-N-oxy group), ammonium salt (amine salt), hydrazine group, disulfide/disulfide compound (such as linear O-dithiopyridine, etc., such as cyclic lipoic acid, etc.), C17 (ester group, thioester group), C18 (carbonate group, thiocarbonate group, dithiocarbonate group, trithiocarbonate group/trithiocarbonate group), C19 (hydroxylamine amide group), xanthate group, peroxythiocarbonate group, tetrathiodiester group, O-carbonylhydroxylamino group, amide group, imide group, hydrazide group, sulfonyl hydrazide group, hydrazone group, imine group, hydrazine group, or the like, An enamine group, an alkynylamine group, a protected hydroxyl group or mercapto group (carbamate, monothiocarbamate, dithiocarbamate), a protected amino group (carbamate, monothiocarbamate, dithiocarbamate), or the like;
class D: carboxyl group, sulfonic acid group, sulfenic acid group, hydroxamic acid group, thiohydroxamic acid group, xanthic acid group, acid halide, sulfonyl chloride group, aldehyde group, glyoxal group, acetal group, hemiacetal group, hydrated aldehyde group, ketone group, ketal group, hemiketal group, ketal group, hydrated ketone group, orthoacid group, orthoester group, cyanate group, thiocyanate group, isonitrile group, isothiocyanate group, ester group, oxycarbonylhalide group, dihydrooxazolyl group (oxazoline D13, isoxazoline), thioaldehyde group, thioketone group, thioacetal group, thioketone hydrate group, thioketal group, thioester group (e.g., D26), thioester group (e.g., D27), dithioester group (e.g., D18), thiohemiacetal group, monothiohydrate group, dithiohydrate group, thiol hydrate group, thiocarboxylic acid group [ monothiocarboxylic acid (thiocarbonyl D16 or thiohydroxy D15) ] Dithiocarboxylic acid group D17], urea group, thiourea group, guanidine group and protonated form thereof, amidine group and protonated form thereof, acid anhydride group, squaric acid ester group, hemisquaric acid group, N-carbamoyl-3-imidazolyl group, N-carbamoyl-3-methylimidazolium iodide, imido group, nitrone group, oxime group, urea group, thiourea group, pseudourea group and the like;
Class E: maleimide group, acrylate group, N-acrylamide group, methacrylate group, N-methacrylamide group, protected maleimide group (e.g., E5), maleamidic acid group, 1,2, 4-triazoline-3, 5-diketo group, azo group (e.g., linear azo compound, cyclic E7, etc.), cycloalkene group (e.g., cyclooctene group, norbornene group, 7-oxa-bicyclo [2.2.1 ] group]Hept-5-en-2-yl, bicycloheptadiene/2, 5-norbornadiene, 7-oxabicycloheptadienyl, etc.), etc., wherein W in E13 3 Including but not limited to halogen, PhS-, etc. leaving groups;
class F: epoxy groups (glycidyl ether groups), alkenyl groups (including vinyl groups, propenyl groups, etc.), alkenyl hydrocarbon groups (such as allyl groups, etc.), alkynyl groups (such as propynyl groups), alkynyl hydrocarbon groups (such as propargyl groups), etc.;
the class of the signal is a class G,
class Ga: cycloalkynyls or cycloalkynheteroalkyls (e.g., G1, G2, G3, G4, G7, G8, G9, G10), conjugated dienyls (e.g., linear butadienyl, such as cyclic cyclopentadiene), hybrid conjugated dienyl (e.g., furyl), 1,2,4, 5-tetrazinyl, etc.;
class Gb: azido, a nitrile oxide group, a cyanooxide group, a cyano group, an isocyano group, an aldoximo group, a diazo group, a diazonium ion, an azoxy group, a nitrilo imino group, an aldimino-N-oxide group, a tetrazolo group, a 4-acetyl-2-methoxy-5-nitrophenoxy group (G31), and a diazotized form thereof (G32); other functional groups capable of undergoing 1, 3-dipolar cycloaddition are also included in the present invention;
Class H: a hydroxyl group (including but not limited to alcoholic hydroxyl, phenolic hydroxyl, enolic hydroxyl, hemiacetal hydroxyl, etc.), a protected hydroxyl group, a siloxy group, a protected dihydroxy group, a trihydroxysilyl group, a protected trihydroxysilyl group, etc.;
functional groups related to the click reaction reported in and cited in adv.funct.mater, 2014,24,2572 are incorporated herein by reference. CN is its oxidized form C ≡ N + O - A precursor of (2), -NH 2 Is ammonium ion-NH 3 + Amine salt-NH 2 HCl precursor-COOH is its sodium salt-COONa, anion-COO — G25 and G26 are precursors of each other, G5 and G6 are precursors of G2 and G3, respectively, and G31 is a precursor of G32, and the like. Protected forms include, but are not limited to, protected hydroxyl (e.g., H2), protected dihydroxy (e.g., H3), protected trihydroxy (e.g., H5), protected orthocarbonic acid (D8), protected thiol (e.g., C2), protected amino (e.g., C6, C16), protected carboxy (e.g., D11), protected aldehyde (e.g., D7), protected maleimide (e.g., E4), protected alkynyl (e.g., F4), and the like. Substituted forms are also included in A13, A14, E9-E12. -NH (C ═ NH) 2 + )NH 2 Is a protonated form of the guanidino group. One functional group can belong to both sub-classes simultaneously. The o-pyridine disulfide of C13 also belongs to Protected forms of sulfhydryl groups. C9 is both a protected amino group and a protected dihydroxy H3. Esters, thioesters and carbonates or thiocarbonates of C17 and C18 also belong to the group of protected hydroxy or mercapto groups.
1.6.2. The use of the above functional groups (including variations thereof) includes, by way of example, but is not limited to:
groups of class a can be modified with amino groups to form amide or carbamate linkages.
The sulfonic acid ester or sulfinic acid ester in the group of the B can be used for alkylation modification, and the group containing a sulfone group or a sulfoxide group can be used for modification of a sulfydryl group or a disulfide bond.
C-like groups are also frequently present at modified sites of biologically relevant substances, such as sulfhydryl groups, amino groups, disulfide bonds, etc. This class is mainly composed of groups with similar reactivity (e.g., hydroxylamine, hydrazine), protected forms, salt forms, etc., and further includes readily removable halogens, etc. C10 such as iodoacetamide may also be modified with a thiol group. C13 and C14 may also belong to the protected mercapto group C3. Typical examples of C14 are lipoic acid.
Groups of class D or deprotected forms may be reactive with hydroxyl groups or groups of class C, such as D1-D6, D9, D10, D12, D13, D14-D16, D19, D20, D21, D22, D23, D25, D29, or deprotected forms of D7, D8, D11, D18, D24, D26-D28, with appropriate groups of amino, mercapto, hydroxyl, or halo. Groups in class D may also react with other groups in this class, for example D25 may react with D1 and D13 may react with D1, D4. Wherein, the guanidyl can form a dihydrobond with two carbonyl groups of the tanshinone IIa.
Groups of class E contain α, β -unsaturation and can undergo 1, 2-addition reactions, for example with amino groups in class C, mercapto groups, and hydroxyl groups in class H. E13 can also undergo a substitution reaction with a dimercapto group.
The F-like group, the most common structures of which have similarity in preparation method, can be obtained by substitution reaction of corresponding halides. The epoxy group includes, but is not limited to, a dihydroxy group exposed by ring opening, a ring-opening addition reaction with an amino group, and the like. The alkenyl group of F2 may undergo an addition reaction. F3 and deprotected F4 are common groups for click reactions.
Groups of the class G can carry out click reaction and are divided into two subclasses of Ga and Gb, cycloalkyne and precursor thereof in Ga, conjugated diene and 1,2,4, 5-tetrazinyl can carry out cycloaddition reaction or Diels-Alder addition reaction, and allyl, propargyl, allene and other groups in Gb can carry out 1, 3-dipolar cycloaddition reaction. In addition, G31 can be converted into a reactive group shown as G32 by diamine treatment or the like, and G32 can react with a carboxyl group to form an ester bond.
H-like groups are hydroxyl, dihydroxy, trihydroxy, or any protected form, and are important functional modifications of the present invention as the starting group (e.g., from the PEG terminus), and groups containing hydroxyl groups or their deprotonated forms are also essential components of the initiator center for initiating polymerization of ethylene oxide in the present invention. The hydroxyl group in class H may also be present at the modified site of the biologically relevant substance. In addition, H6, H7 can be converted to enolic hydroxyl under light conditions, which in turn can undergo addition reactions with α, β -unsaturated bonds as in class E.
R 01 Or does not have a bonding reaction with the biologically relevant substance, and has a specific function including both the functional groups of the targeting group and the fluorescent group or a substituted form thereof. The substituted forms need to still have the corresponding special function and can be classified as corresponding targeting groups and fluorescent groups. Such R 01 Including but not limited to class I to class J:
class I: targeting groups and pharmaceutically acceptable salts thereof, such as folic acid and derivatives thereof, cholesterol and derivatives thereof, biotin and derivatives thereof, and the like. Derivatives of biotin such as D-desthiobiotin, 2-iminobiotin and the like.
Class J: examples of the photosensitive group or the fluorescent group include phthalocyanine complexes, fluorescein, rhodamine, anthracene, pyrene, coumarin, fluorescein 3G, carbazole, imidazole, indole, alizarin violet, and any of the above functional derivatives. The derivatives of rhodamine include, but are not limited to, tetramethylrhodamine, tetraethylrhodamine (rhodamine B, RB200), rhodamine 3G, rhodamine 6G (rhodamine 590), 5-carboxy-X-rhodamine, 6-carboxy-X-rhodamine, sulforhodamine B, sulforhodamine G, sulforhodamine 101, rhodamine X (R101), rhodamine 101, rhodamine 110, rhodamine 123, rhodamine 700, rhodamine 800, and the like, and also include, but are not limited to, rhodamine derivatives described in the literature { Progress in Chemistry,2010,22(10):1929-1939} and citations thereof.
In the present invention, - (Z) 1 ) q1 -R 01 Functional groups as a whole. Wherein, for example, R 01 Functional groups include, but are not limited to, groups described and exemplified in documents CN104877127A, CN104530413A, CN104530415A, CN104530417A, including, but not limited to, CN104530417A paragraph [0423 ], active ester, amino group, aldehyde group, carboxyl group, acid halide, acid anhydride, cyano group, alkynyl group, hydroxyl group, and the like]～[0432]Section CN104877127A [0308]～[0328]For example, they are not described in detail herein.
1.6.3. Functional group R 01 Structural classification of
Specifically, R 01 Including but not limited to functional groups in any of classes A through J, variations of classes A through H, functional derivatives of class I through J; the variant is selected from any one of a precursor of a reactive group, an active form thereof as a precursor, a substituted active form, a protected form, a deprotected form:
class A:
Figure BDA0002963906360000271
Figure BDA0002963906360000281
or class B:
Figure BDA0002963906360000282
or class C:
Figure BDA0002963906360000283
or class D:
Figure BDA0002963906360000284
or class E:
Figure BDA0002963906360000291
or class F:
Figure BDA0002963906360000292
or class G: class Ga:
Figure BDA0002963906360000293
or class Gb:
Figure BDA0002963906360000294
or class H:
Figure BDA0002963906360000301
or class I:
Figure RE-GDA0003038345320000302
or class J:
Figure BDA0002963906360000303
Figure BDA0002963906360000311
and the like.
Wherein Q 5 、M 5 And M 5 The rings are consistent with the above definitions and are not described further herein.
Wherein, Y 1 Is a leaving group attached to a sulfonyl, sulfinyl, oxysulfonyl or oxysulfinyl group. Y is 1 There is no particular limitation. Y is 1 Preferably having C 1-10 Hydrocarbyl or fluoro C 1-10 A hydrocarbyl group. Y is 1 More preferably having C 1-10 Alkyl radical, C 1-10 Alkenyl, phenyl, and the like, or substituted forms thereof. Wherein, the substituted atom or the substituted group is halogen atom, alkenyl, alkoxy or nitro. Specifically, as an example Y 1 Can be selected from any one of the group including, but not limited to, methyl, ethyl, n-propyl, isopropyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, vinyl, phenyl, benzyl, p-methylphenyl, 4- (trifluoromethoxy) phenyl, trifluoromethyl, 2,2, 2-trifluoroethyl, and the like. Wherein, butyl includes but is not limited to n-butyl and tert-butyl. Octyl includes, but is not limited to, n-octyl, 2-ethylhexyl. Y is 1 More preferably, it is any of methyl, p-methylphenyl, 2,2, 2-trifluoroethyl, trifluoromethyl, vinyl, and the like.
Wherein W is F, Cl, Br or I, preferably Br or Cl.
Wherein, W 3 Is a leaving group including, but not limited to, F, Cl, Br, I, PhS-, preferably Br or Cl.
Wherein, W 2 Is F, Cl, Br or I, preferably I.
Wherein the content of the first and second substances,
Figure BDA0002963906360000312
Figure BDA0002963906360000313
are respectively a ringCyclic structures containing nitrogen atoms, azonium ions, double bonds, azo, triple bonds, disulfide bonds, anhydrides, imides, dienes in the backbone, including but not limited to carbocycles, heterocycles, benzoheterocycles, substituted carbocycles, substituted heterocycles or substituted benzoheterocycles, and the like.
Wherein M is a carbon or heteroatom in the ring, including but not limited to carbon, nitrogen, phosphorus, silicon.
Wherein, M 8 Is a carbon atom or a heteroatom located on a ring. M 8 Preferably a carbon atom, a nitrogen atom, a phosphorus atom or a silicon atom. M 8 The number of ring-forming atoms of the ring is not particularly limited, but is preferably 4 to 50, more preferably 4 to 32, still more preferably 5 to 18, and most preferably 5 to 8. M 8 Can be a carbon atom or a heteroatom in a 4-50 membered ring, preferably a carbon atom, a nitrogen atom, a phosphorus atom or a silicon atom in a 4-32 membered ring, more preferably a carbon atom, a nitrogen atom, a phosphorus atom or a silicon atom in a 5-18 membered ring; most preferably a carbon atom, a nitrogen atom, a phosphorus atom or a silicon atom on a 5-to 8-membered ring.
Wherein M is 22 Is a carbon atom or a hetero atom on an alicyclic or alicyclic ring, and may be selected from a carbon atom, a nitrogen atom, a phosphorus atom or a silicon atom. M 22 The number of ring-forming atoms of the ring is 4, 5, 6, 7 or 8, preferably 4, 5 or 6.
Wherein PG 8 Is a protecting group for orthocarbonic acid or orthosilicic acid, D8 is a protected form of orthosilicic acid, H5 is a protected form of orthosilicic acid. PG (PG) 8 May be a single trivalent end group such as
Figure BDA0002963906360000314
Taking D8 as an example, corresponding to
Figure BDA0002963906360000315
PG 8 It may also be two or three separate end groups, and D8 corresponds to
Figure BDA0002963906360000316
H5 corresponds to
Figure BDA0002963906360000317
Wherein R is 2 Is an end group or a divalent linking group connecting oxygen or sulfur atoms in an acetal, ketal, hemiacetal, hemiketal, orthoester, thioacetal, thioketal, thiohemiacetal, thiohemiketal, thioorthoester, orthosilicate ester and the like structure, such as D7, D18, D8 and H5.
R 2 Can be selected from hydrogen atoms, R 21 Or R 3 Any one atom or group.
Wherein R is 21 Is a divalent linking group and participates in ring formation.
R 21 The number of carbon atoms of (A) is not particularly limited, but is preferably 1 to 20, more preferably 1 to 10.
R 21 The structure of (a) is not particularly limited and includes, but is not limited to, a linear structure, a branched structure containing a side group, or a cyclic structure. Among them, the cyclic structure is not particularly limited, and the above-mentioned aliphatic ring, aromatic ring, sugar ring, and condensed ring are preferable.
R 21 May or may not contain heteroatoms.
R 21 Is selected from C 1-20 Alkylene, divalent C 1-20 Heterohydrocarbyl, substituted C 1-20 Alkylene, substituted divalent C 1-20 Any one divalent linking group or any two or any three of divalent linking groups in the heterohydrocarbon group. Wherein, the substituent atom or substituent group is not particularly limited, including but not limited to any substituent atom or any substituent group of the term part, selected from any one of halogen atom, alkyl substituent group, and heteroatom-containing substituent group.
R 21 Preferably C 1-20 Open-chain alkylene, C 1-20 Alkenyl radical, C 1-20 Cycloalkylene radical, C 1-20 Cycloalkylene, arylene, divalent C 1-20 Aliphatic heteroalkyl, divalent C 1-20 Aliphatic heteroalkenyl, divalent heteroaryl, divalent heteroarylalkyl, substituted alkylene, substituted C 1-20 Open alkenylene, substituted C 1-20 Cycloalkylene, substituted C 1-20 Cycloalkylene radical, substituted arylene radical, substituted divalent C radical 1-20 Lipoheteroalkyl, substituted divalent C 1-20 Any one of divalent linking groups of lipoheteroalkenyl, substituted divalent heteroaryl, substituted divalent heteroarylalkyl, a combination of any two, or a combination of any three. Among them, the substituent atom or the substituent is preferably a halogen atom, an alkoxy group and a nitro group.
R 21 More preferably C 1-10 Open-chain alkylene, C 1-10 Alkenyl radical, C 3-10 Cycloalkylene radical, C 1-10 Cycloalkylene radical, arylene radical, divalent C 1-10 Aliphatic heteroalkyl, divalent C 1-10 Lipoheteroalkenyl, divalent heteroaryl, divalent heteroarylalkyl, substituted alkylene, substituted C 1-10 Open alkenylene, substituted C 1-10 Cycloalkylene, substituted C 1-10 Cycloalkylene radical, substituted arylene radical, substituted aralkylene radical, substituted divalent C 1-10 Lipoheteroalkyl, substituted divalent C 1-10 Any one of divalent linking groups of the lipoheteroalkenyl group, the substituted divalent heteroaryl group and the substituted divalent heteroaryl hydrocarbon group, any two of the divalent linking groups or any three of the divalent linking groups.
Specifically, R 21 Selected from the group consisting of methylene, ethylene, propylene, butylene, pentylene, hexylene, heptylene, octylene, nonylene, decylene, 1, 2-phenylene, benzylene, C 1-20 Oxaalkylene, C 1-20 Thiaalkylene group, C 1-20 Any one group of azaalkylene and azaaralkyl, substituted forms of any one group, and any two or more of the same or different groups or substituted forms thereof. Among them, the substituent atom or the substituent is selected from any one of a halogen atom, a hydrocarbon substituent and a substituent containing a hetero atom, and a halogen atom, an alkoxy group or a nitro group is preferable.
R 21 More preferred are 1, 2-ethylene group and 1, 3-propylene group.
Wherein R is 3 To connect oxygenRadicals or sulfur radicals.
R 3 The number of carbon atoms of (A) is not particularly limited, but is preferably 1 to 20, more preferably 1 to 10.
R 3 The structure of (a) is not particularly limited and includes, but is not limited to, a linear structure, a branched structure containing a side group, or a cyclic structure. Among them, the cyclic structure is not particularly limited, and the above-mentioned aliphatic ring, aromatic ring, sugar ring, and condensed ring are preferable.
R 3 May or may not contain heteroatoms.
R 3 Is selected from C 1-20 Hydrocarbyl radical, C 1-20 Heterohydrocarbyl radical, C 1-20 Substituted hydrocarbyl radical, C 1-20 Any of substituted heterohydrocarbyl groups. For substitution of R 3 The heteroatom or substituent of (a) is not particularly limited and includes, but is not limited to, any heteroatom or any substituent of the term moiety, preferably any one selected from a halogen atom, a hydrocarbyl group, a heteroatom-containing substituent.
R 3 Preferably C 1-20 Alkyl radical, C 3-20 Alkylene, aryl, C 1-20 Aliphatic heterocarbyl, heteroaryl, heteroaromatic hydrocarbyl, substituted C 1-20 Alkyl, substituted C 3-20 Alkylene, substituted aryl, substituted aralkyl, substituted C 1-20 Any one of aliphatic heterocarbon group, substituted heteroaryl group and substituted heteroaromatic hydrocarbon group. Wherein the substituent atom or substituent is selected from any one of halogen atom, alkyl substituent and heteroatom-containing substituent.
R 3 Preferably C 1-20 Straight chain alkyl, C 1-20 Branched alkyl radical, C 3-20 Cycloalkyl, aryl, aralkyl, C 1-20 Aliphatic heterocarbyl, heteroaryl, heteroaromatic hydrocarbyl, substituted C 1-20 Straight chain alkyl, substituted C 1-20 Branched alkyl, substituted C 3-20 Cycloalkyl, substituted aryl, substituted aralkyl, substituted C 1-20 Any one of aliphatic heterocarbon group, substituted heteroaryl group and substituted heteroaromatic hydrocarbon group. Wherein the substituent atom or substituent is selected from any one of a halogen atom, a hydrocarbon substituent and a heteroatom-containing substituent, preferably Selected from halogen atoms, alkoxy, alkyl, aryl or nitro.
R 3 More preferably C 1-10 Straight chain alkyl, C 1-10 Branched alkyl radical, C 3-10 Cycloalkyl, aryl, aralkyl, C 1-20 Aliphatic heterocarbyl, heteroaryl, heteroaromatic hydrocarbyl, substituted C 1-10 Straight chain alkyl, substituted C 1-10 Branched alkyl, substituted C 3-10 Cycloalkyl, substituted aryl, substituted arylalkyl, substituted C 1-10 Any one of aliphatic heterocarbon group, substituted heteroaryl group and substituted heteroaromatic hydrocarbon group. Wherein, the substituent atom or substituent group is selected from any one of halogen atom, alkyl substituent group and substituent group containing hetero atom, preferably fluorine atom, chlorine atom, bromine atom, iodine atom, alkyl, aryl or nitro; more preferably a halogen atom, an alkoxy group or a nitro group.
Specifically, R 3 Selected from any one or any one of methyl, ethyl, n-propyl, isopropyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, benzyl, allyl and the like. Wherein, butyl includes but is not limited to n-butyl and tert-butyl. Octyl includes, but is not limited to, n-octyl, 2-ethylhexyl. Wherein, the substituent atom or substituent group is selected from any one of halogen atom, alkyl substituent group and substituent group containing hetero atom, preferably fluorine atom, chlorine atom, bromine atom, iodine atom, alkyl, aryl or nitro; more preferably a halogen atom, an alkoxy group or a nitro group.
R 3 Most preferred is methyl, ethyl or benzyl.
Wherein R is 4 Is- (R) 4 )C＝N + ＝N — Or- (R) 4 )C - -N + A hydrogen atom, a substituent atom or a substituent on C in the structure of [ identical to ] N.
When taken as a substituent atom, R 4 Selected from any one of halogen atoms. Fluorine atoms are preferred.
When taken as a substituent, R 4 The number of carbon atoms of (A) is not particularly limited, but is preferably 1 to 20, more preferably 1 to 10.
When taken as a substituent, R 4 Is of a structureWith specific limitations including, but not limited to, linear structures, branched structures containing pendant groups, or cyclic structures. Among them, the cyclic structure is not particularly limited, and the above-mentioned aliphatic ring, aromatic ring, sugar ring, and condensed ring are preferable.
When taken as a substituent, R 4 May or may not contain heteroatoms.
R 4 Selected from hydrogen atoms, halogen atoms, C 1-20 Hydrocarbyl radical, C 1-20 Heterohydrocarbyl, substituted C 1-20 Hydrocarbyl or substituted heterohydrocarbyl. Wherein R is 4 The substituent atom or substituent in (1) is not particularly limited, and includes, but is not limited to, any substituent atom or any substituent group of the term moiety selected from any one of a halogen atom, a hydrocarbon group substituent, and a heteroatom-containing substituent.
R 4 More preferably a hydrogen atom, a halogen atom, C 1-20 Alkyl radical, C 1-20 Unsaturated aliphatic radical, aryl radical, aromatic radical, C 1-20 Heterohydrocarbyl radical, C 1-20 Hydrocarbyloxyacyl group, C 1-20 Hydrocarbyl thioacyl, C 1-20 Any one atom or group of hydrocarbyl aminoacyl, or substituted version of any one group. Wherein R is 4 The acyl group in (1) is not particularly limited, including but not limited to any acyl type of the term moiety. R is 4 The acyl group in (1) is more preferably a carbonyl group or a thiocarbonyl group.
R 4 More preferably a hydrogen atom, a halogen atom, C 1-20 Alkyl radical, C 1-20 Alkenyl, aryl, arylalkyl, C 1-20 Aliphatic heteroalkyl, heteroaryl, heteroarylalkyl, C 1-20 Alkoxyacyl, aryloxyacyl, C 1-20 Alkylthio acyl, arylthio acyl, C 1-20 Any one of an alkylaminoacyl group, an arylaminoacyl group, or a substituted version of any one of the groups.
R 4 More preferably a hydrogen atom, a halogen atom, C 1-20 Alkyl radical, C 1-20 Alkenyl, aryl, arylalkyl, C 1-20 Aliphatic heterocarbyl, heteroaryl, heteroaromatic hydrocarbyl, C 1-20 Alkoxycarbonyl, aryloxycarbonyl, C 1-20 Alkyl radicalThiocarbonyl, arylthiocarbonyl, C 1-20 Alkylaminocarbonyl, arylaminocarbonyl, C 1-20 Alkoxythiocarbonyl, aryloxylthiocarbonyl, C 1-20 Alkylthio thiocarbonyl, arylthio thiocarbonyl, C 1-20 Any one atom or group of an alkylaminothiocarbonyl group, arylaminothiocarbonyl group, or a substituted version of any one group.
Specifically, R 4 Selected from the group consisting of, but not limited to, a hydrogen atom, a fluorine atom, a chlorine atom, a bromine atom, an iodine atom, a methyl group, an ethyl group, an n-propyl group, an isopropyl group, a butyl group, a pentyl group, a hexyl group, a heptyl group, an octyl group, a nonyl group, a decyl group, an undecyl group, a dodecyl group, a tridecyl group, a tetradecyl group, a pentadecyl group, a hexadecyl group, a heptadecyl group, an octadecyl group, a nonadecyl group, an eicosyl group, an allyl group, an propenyl group, a vinyl group, a phenyl group, a methylphenyl group, a butylphenyl group, a benzyl group, a methoxycarbonyl group, an ethoxycarbonyl group, a phenoxycarbonyl group, a benzyloxycarbonyl group, a methylthiocarbonyl group, an ethylthiocarbonyl group, an ethoxythiocarbonyl group, a phenoxythiocarbonyl group, a benzyloxythiocarbonyl group, a methylthiothiocarbonyl group, an ethylthiocarbonyl group, a phenylthiocarbonyl group, a, Benzylthiothiocarbonyl, ethylaminothiocarbonyl, benzylaminothiocarbonyl, substituted C 1-20 Alkyl, substituted C 1-20 Alkenyl, substituted aryl, substituted arylalkyl, substituted C 1-20 Aliphatic heterocarbyl, substituted heteroaryl, substituted heteroarylalkyl, substituted C 1-20 Alkoxycarbonyl, substituted aryloxycarbonyl, substituted C 1-20 Alkylthio carbonyl, substituted arylthio carbonyl, substituted C 1-20 Alkylaminocarbonyl, substituted arylaminocarbonyl, substituted C 1-20 Alkoxythiocarbonyl, substituted aryloxythiocarbonyl, substituted C 1-20 Alkylthio thiocarbonyl, substituted arylthio thiocarbonyl, substituted C 1-20 An alkylaminothiocarbonyl group, a substituted arylaminothiocarbonyl group, or the like. Wherein, butyl includes but is not limited to n-butyl and tert-butyl.Octyl includes, but is not limited to, n-octyl, 2-ethylhexyl. The substituent atom or substituent is selected from any one of a halogen atom, a hydrocarbon substituent and a heteroatom-containing substituent, and is preferably a fluorine atom, a chlorine atom, a bromine atom, an iodine atom, an alkenyl group or a nitro group.
R 4 More preferred is a hydrogen atom, methyl group, ethyl group, n-propyl group, isopropyl group, butyl group, pentyl group, hexyl group, heptyl group, octyl group, nonyl group, decyl group, allyl group, propenyl group, vinyl group, phenyl group, methylphenyl group, butylphenyl group, benzyl group, methoxycarbonyl group, ethoxycarbonyl group, phenoxycarbonyl group, benzyloxycarbonyl group, methylthiocarbonyl group, ethylthiocarbonyl group, phenylthiocarbonyl group, benzylthiocarbonyl group, ethylaminocarbonyl group, benzylaminocarbonyl group, methoxythiocarbonyl group, ethoxythiocarbonyl group, phenoxythiocarbonyl group, benzyloxythiocarbonyl group, methylthiothiocarbonyl group, ethylthiocarbonyl group, phenylthiocarbonyl group, benzylthiocarbonyl group, ethylaminothiocarbonyl group, benzylaminothiocarbonyl group, C 1-10 Halogenated hydrocarbon group, halogenated phenyl, halogenated benzyl, nitro phenyl and any kind of atom or group, or any kind of substituted form of group.
R 4 Preferably any one atom or group of hydrogen atom, methyl group, ethyl group, n-propyl group, isopropyl group, butyl group, pentyl group, hexyl group, heptyl group, octyl group, nonyl group, decyl group, allyl group, propenyl group, vinyl group, phenyl group, methylphenyl group, butylphenyl group, and benzyl group.
R 4 Most preferably a hydrogen atom, a methyl group or a benzyl group.
Wherein R is 8 、R 9 、R 10 、R 11 、R 12 Each independently is a hydrogen atom, a substituent atom or a substituent on a double bond (-C-), and R is in the same molecule 8 、R 9 、R 10 、R 11 、R 12 May be the same as or different from each other.
When it is a substituted atom, R 8 、R 9 、R 10 、R 11 、R 12 Each independently selected from any one halogen atom of F, Cl, Br and I. Each independently preferably being a fluorine atom.
When it is a substituent, R 8 、R 9 、R 10 、R 11 、R 12 The number of carbon atoms of (a) is not particularly limited. R 8 、R 9 、R 10 、R 11 、 R 12 The number of carbon atoms of (A) is preferably 1 to 20, more preferably 1 to 10, independently of each other.
When it is a substituent, R 8 、R 9 、R 10 、R 11 、R 12 The structure of (a) is not particularly limited, and each independently includes, but is not limited to, a linear structure, a branched structure containing a pendant group, or a cyclic structure. Among them, the cyclic structure is not particularly limited, and the above aliphatic ring, aromatic ring, sugar ring, and condensed ring are preferable.
When it is a substituent, R 8 、R 9 、R 10 、R 11 、R 12 Each independently may or may not contain a heteroatom.
R 8 、R 9 、R 10 、R 11 、R 12 Each independently selected from a hydrogen atom, a halogen atom, C 1-20 Hydrocarbyl radical, C 1-20 Heterohydrocarbyl, substituted C 1-20 Hydrocarbyl or substituted heterohydrocarbyl. Wherein R is 8 The substituent atom or substituent in (1) is not particularly limited, and includes, but is not limited to, any substituent atom or any substituent group of the term moiety, selected from any one of a halogen atom, a hydrocarbyl substituent group, and a heteroatom-containing substituent group.
R 8 、R 9 、R 10 、R 11 、R 12 Each independently more preferably a hydrogen atom, a halogen atom, C 1-20 Alkyl radical, C 1-20 Unsaturated aliphatic, aryl, C 1-20 Heterohydrocarbyl radical, C 1-20 Hydrocarbyloxyacyl group, C 1-20 Hydrocarbyl thioacyl, C 1-20 Any atom or group of a hydrocarbylaminoacyl group, or a substituted version of any group. Wherein R is 8 The acyl group in (1) is not particularly limited, including but not limited to any acyl type of the term moiety.
R 8 、R 9 、R 10 、R 11 、R 12 Each independently more preferably a hydrogen atom, a halogen atom, C 1-20 Alkyl radical, C 1-20 Alkenyl, aryl, arylalkyl, C 1-20 Aliphatic heterocarbyl, heteroaryl, heteroaromatic hydrocarbyl, C 1-20 Alkoxyacyl, aryloxyacyl, C 1-20 Alkylthio acyl, arylthio acyl, C 1-20 Any one atom or group of an alkylaminoacyl group, an arylaminoacyl group, or a substituted version of any one group. The substituent atom or substituent is selected from any one of halogen atom, alkyl substituent and hetero atom-containing substituent, preferably halogen atom, alkenyl or nitro
R 8 、R 9 、R 10 、R 11 、R 12 Each independently more preferably a hydrogen atom, a halogen atom, C 1-20 Alkyl radical, C 1-20 Alkenyl, aryl, arylalkyl, C 1-20 Aliphatic heteroalkyl, heteroaryl, heteroarylalkyl, C 1-20 Alkoxycarbonyl, aryloxycarbonyl, C 1-20 Alkylthio carbonyl, arylthio carbonyl, C 1-20 Alkylaminocarbonyl, arylaminocarbonyl, C 1-20 Alkoxythiocarbonyl, aryloxylthiocarbonyl, C 1-20 Alkylthio thiocarbonyl, arylthio thiocarbonyl, C 1-20 Any one atom or group of an alkylaminothiocarbonyl group, arylaminothiocarbonyl group, or a substituted version of any one group. R 8 The acyl group in (1) is more preferably a carbonyl group or a thiocarbonyl group. Wherein, the substituent atom or substituent is selected from any one of halogen atom, alkyl substituent and substituent containing heteroatom, preferably fluorine atom, chlorine atom, bromine atom, iodine atom, alkenyl or nitro.
In particular, R 8 、R 9 、R 10 、R 11 、R 12 Each independently selected from the group consisting of, but not limited to, hydrogen atom, fluorine atom, chlorine atom, bromine atom, iodine atom, methyl group, ethyl group, n-propyl group, isopropyl group, butyl group, pentyl group, hexyl group, heptyl group, octyl group, nonyl group, decyl group, undecyl group, dodecyl group, tridecyl group, tetradecyl group, pentadecyl group, hexadecyl group, heptadecyl group, octadecyl group A group, nonadecyl group, eicosyl group, allyl group, propenyl group, vinyl group, phenyl group, methylphenyl group, butylphenyl group, benzyl group, methoxycarbonyl group, ethoxycarbonyl group, phenoxycarbonyl group, benzyloxycarbonyl group, methylthiocarbonyl group, ethylthiocarbonyl group, phenylthiocarbonyl group, benzylthiocarbonyl group, ethylaminocarbonyl group, benzylaminocarbonyl group, methoxythiocarbonyl group, ethoxythiocarbonyl group, phenoxythiocarbonyl group, benzyloxythiocarbonyl group, methylthiothiocarbonyl group, ethylthiothiocarbonyl group, phenylthiothiocarbonyl group, benzylthiocarbonyl group, ethylaminothiocarbonyl group, benzylaminothiocarbonyl group, substituted C 1-20 Alkyl, substituted C 1-20 Alkenyl, substituted aryl, substituted arylhydrocarbon, substituted C 1-20 Aliphatic heterocarbyl, substituted heteroaryl, substituted heteroarylalkyl, substituted C 1-20 Alkoxycarbonyl, substituted aryloxycarbonyl, substituted C 1-20 Alkylthio carbonyl, substituted arylthio carbonyl, substituted C 1-20 Alkylaminocarbonyl, substituted arylaminocarbonyl, substituted C 1-20 Alkoxythiocarbonyl, substituted aryloxythiocarbonyl, substituted C 1-20 Alkylthio thiocarbonyl, substituted arylthio thiocarbonyl, substituted C 1-20 An alkylaminothiocarbonyl group, a substituted arylaminothiocarbonyl group, or the like. Wherein, butyl includes but is not limited to n-butyl and tert-butyl. Octyl includes, but is not limited to, n-octyl, 2-ethylhexyl. Among them, the substituent atom or the substituent is selected from any one of a halogen atom, a hydrocarbon substituent and a heteroatom-containing substituent, and is preferably a halogen atom, an alkenyl group or a nitro group.
R 8 、R 9 、R 10 、R 11 、R 12 Further preferred are, independently of one another, a hydrogen atom, methyl group, ethyl group, n-propyl group, isopropyl group, butyl group, pentyl group, hexyl group, heptyl group, octyl group, nonyl group, decyl group, allyl group, propenyl group, vinyl group, phenyl group, methylphenyl group, butylphenyl group, benzyl group, methoxycarbonyl group, ethoxycarbonyl group, phenoxycarbonyl group, benzyloxycarbonyl group, methylthiocarbonyl group, ethylthiocarbonyl group, phenylthiocarbonyl group, benzylthiocarbonyl group, ethylaminocarbonyl group, benzylaminocarbonyl groupA group selected from the group consisting of methoxythiocarbonyl, ethoxythiocarbonyl, phenoxythiocarbonyl, benzyloxythiocarbonyl, methylthiothiocarbonyl, ethylthiothiocarbonyl, phenylthiothiocarbonyl, benzylthiocarbonyl, ethylaminothiocarbonyl, benzylaminothiocarbonyl and C 1-10 Halogenated hydrocarbon group, halogenated phenyl, halogenated benzyl, nitro phenyl and any kind of atom or group, or any kind of substituted form of group. The substituent atom or substituent is selected from any one of a halogen atom, a hydrocarbon substituent and a heteroatom-containing substituent, and is preferably a fluorine atom, a chlorine atom, a bromine atom, an iodine atom, an alkenyl group or a nitro group.
R 8 、R 9 、R 10 、R 11 、R 12 Each independently is more preferably a hydrogen atom, a fluorine atom or a methyl group.
In class E3, R 8 Most preferred is methyl.
Wherein R is 24 For attachment to the end groups of disulfide bonds, C is preferred 1-20 Alkyl, aryl, heteroaryl, and the like, such as ortho-pyridyl.
Wherein R is 27 For the substituent attached to the azo, a phenyl group, a substituted phenyl group or a hybridized phenyl group is preferred.
Wherein R is 30 Is a hydrocarbon radical, preferably C 1-20 Alkyl, benzyl, phenyl ring hydrogen atoms by C 1-20 Hydrocarbyl-substituted benzyl.
Wherein M is 19 、M 20 、M 21 Each independently an oxygen atom or a sulfur atom, and may be the same as or different from each other in the same molecule.
Wherein, X 11 To attach a carbonyl or thiocarbonyl end group, preferably C 1-20 Alkyl groups, more preferably methyl, ethyl, isopropyl, tert-butyl.
Wherein, X 12 Terminal groups to which carbonate or thiocarbonate groups are attached, selected from hydrocarbon groups (which may or may not include a benzene ring), preferably C 1-20 Hydrocarbyl, more preferably C 1-20 Alkyl, phenylhydrocarbyl or hydrocarbyl substituted phenyl.
Wherein, X 13 To a sulfur radicalTerminal group selected from thiol protecting group or group LG 2 。
When it is a mercapto-protecting group, X 13 Is selected from PG 2 Thiol protecting groups in the listed groups.
Wherein LG is 2 The number of carbon atoms of (a) is not particularly limited. LG (Ligno-lead-acid) 2 The number of carbon atoms of (A) is preferably 1 to 20, more preferably 1 to 10.
LG 2 The structure of (a) is not particularly limited and includes, but is not limited to, a linear structure, a branched structure containing a side group, or a cyclic structure. The cyclic structure is not particularly limited, and the above-mentioned aliphatic ring, aromatic ring, sugar ring, and condensed ring are preferable.
LG 2 May or may not contain heteroatoms.
LG 2 Is selected from C 1-20 Hydrocarbyl radical, C 1-20 Heterohydrocarbyl, substituted C 1-20 Any one of a hydrocarbon group and a substituted heterohydrocarbon group. Wherein LG is 2 The substituted heteroatom or substituent in (1) is not particularly limited, including but not limited to any substituted heteroatom or any substituent of the term moiety selected from any of halogen atoms, hydrocarbyl substituents, heteroatom-containing substituents.
LG 2 More preferably C 1-20 Alkyl radical, C 1-20 Unsaturated aliphatic, aryl, C 1-20 Heterohydrocarbyl radical, C 1-20 Alkylthio radical, C 1-20 Aliphatic heterocarbylthio, arylthio, C 1-20 Fatty hydrocarbyl acyl radical, C 1-20 Lipoheteroalkylacyl, arylacyl, heteroarylacyl, C 1-20 Hydrocarbyloxyacyl group, C 1-20 Hydrocarbyl thioacyl, C 1-20 Hydrocarbyl aminoacyl radical, C 1-20 Heterohydrocarbyloxyacyl group, C 1-20 Heterocarbylthioacyl radical, C 1-20 Any one group or substituted version of any one group of a heterocarbylaminoacyl group. Wherein LG is 2 The acyl group in (1) is not particularly limited, including but not limited to any acyl type of the term moiety. By way of example, LG 2 The acyl group in (1) may be selected from the group consisting of a carbonyl group, a sulfonyl group, a sulfinyl group, a phosphoryl group,Nitroxyl, nitrosyl, thiocarbonyl, imidoyl, thiophosphoryl, dithiophosphoryl, trithiophosphoryl, thiophosphorous acyl, dithiophosphoryl, thiophosphinic acyl, thiophosphinic group, thiophosphoryl, dithiophosphono, thiophosphinic group, and the like. Any of acyl groups such as a carbonyl group, a thiocarbonyl group, a sulfonyl group, and a sulfinyl group is preferable. LG (Ligno-lead-acid) 2 The acyl group in (1) is more preferably a carbonyl group, thiocarbonyl group or sulfonyl group.
LG 2 More preferably C 1-20 Alkyl, aryl, aralkyl, C 1-20 Heteroalkyl, heteroaryl, heteroaralkyl, C 1-20 Alkylthio, arylthio, aralkylthio, C 1-20 Heteroalkylthio, heteroarylthio, heteroaralkylthio, C 1-20 Alkylcarbonyl, arylcarbonyl, aralkylcarbonyl, C 1-20 Heteroalkylcarbonyl, heteroarylcarbonyl, heteroaralkylcarbonyl, C 1-20 Alkoxycarbonyl, aryloxycarbonyl, aralkyloxycarbonyl, C 1-20 Alkylthio-carbonyl, arylthio-carbonyl, aralkylthiocarbonyl, C 1-20 Alkylaminocarbonyl, arylaminocarbonyl, aralkylaminocarbonyl, C 1-20 Heteroalkyloxycarbonyl, heteroaryloxycarbonyl, heteroarylalkyloxycarbonyl, C 1-20 Heteroalkylthio-carbonyl, heteroarylthio-carbonyl, heteroaralkylthio-carbonyl, C 1-20 Heteroalkylaminocarbonyl, heteroarylaminocarbonyl, heteroarylalkylaminocarbonyl, C 1-20 Alkylthio, arylthio, aralkylthiocarbonyl, C 1-20 Heteroalkylthiocarbonyl, heteroarylthiocarbonyl, heteroarylalkylthiocarbonyl, C 1-20 Alkoxythiocarbonyl, aryloxylthiocarbonyl, aralkyloxythiocarbonyl, C 1-20 Alkylthio thiocarbonyl, arylthio thiocarbonyl, aralkylthio thiocarbonyl, C 1-20 Alkylaminothiocarbonyl, arylaminothiocarbonyl, aralkylaminothiocarbonyl, C 1-20 Heteroalkyloxythiocarbonyl, heteroaryloxythiocarbonyl, heteroarylalkoxythiocarbonyl, C 1-20 Heteroalkylthio thiocarbonyl, heteroarylthio thiocarbonyl, heteroarylalkylthioSubstituted carbonyl, C 1-20 A heteroalkylaminothiocarbonyl group, a heteroarylaminothiocarbonyl group, a heteroarylalkylaminothiocarbonyl group or a substituted version of any group.
LG 2 More preferably C 1-20 Alkyl, aryl, aralkyl, C 1-20 Heteroalkyl, heteroaryl, heteroaralkyl, C 1-20 Alkylthio, arylthio, aralkylthio, C 1-20 Any one of the groups or substituted versions of any one of the groups heteroarylthio, heteroaralkylthio.
Specifically, LG 2 Selected from the group including but not limited to methyl, ethyl, n-propyl, isopropyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, eicosyl, allyl, trityl, phenyl, benzyl, methylbenzyl, nitrobenzyl, tert-butylthio, benzylthio, 2-pyridylthio, ethylacoyl, phenylformyl, methoxyacyl, ethoxyacyl, tert-butyloxyacyl, phenoxyacyl, benzyloxyacyl, methylthioacyl, ethylthioacyl, tert-butylthioacyl, phenylthioacyl, benzylthioacyl, 2-pyridylcarbonyl, methylaminoacyl, ethylaminoacyl, tert-butylaminoacyl, benzylaminoacyl, and the like, or a substituted form of any of these groups. Wherein, butyl includes but is not limited to n-butyl and tert-butyl. Octyl includes, but is not limited to, n-octyl, 2-ethylhexyl. Among them, the substituent atom or the substituent is selected from any one of a halogen atom, a hydrocarbon group substituent and a hetero atom-containing substituent, and is preferably a fluorine atom, a chlorine atom, a bromine atom, an iodine atom or a nitro group.
LG 2 More preferably, it is methyl, ethyl, n-propyl, isopropyl, tert-butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, eicosyl, allyl, trityl, phenyl, benzylA group, a methylbenzyl group, a nitrobenzyl group, a tert-butylthio group, a benzylthio group, a 2-pyridylthio group, an acetyl group, a benzoyl group, a methoxycarbonyl group, an ethoxycarbonyl group, a tert-butyloxycarbonyl group, a phenoxycarbonyl group, a benzyloxycarbonyl group, a methylthiocarbonyl group, an ethylthiocarbonyl group, a tert-butylthiocarbonyl group, a phenylthiocarbonyl group, a benzylthiocarbonyl group, a 2-pyridylcarbonyl group, a methylaminocarbonyl group, an ethylaminocarbonyl group, a tert-butylaminocarbonyl group, an ethylaminocarbonyl group, an ethylthiocarbonyl group, a phenylthiocarbonyl group, a methylthiothiocarbonyl group, an ethoxythiocarbonyl group, a tert-butyloxythiocarbonyl group, a phenoxythiocarbonyl group, a benzyloxythiocarbonyl group, a methylthiothiocarbonyl group, an ethylthiocarbonyl group, a tert-butylthiocarbonyl group, a phenylthiocarbonyl group, a benzylthiocarbonyl group, a methylaminocarbonyl group, a methylthiocarbonyl group, a phenylthiocarbonyl group, a, Ethylaminothiocarbonyl, tert-butylaminothiocarbonyl, benzylaminothiocarbonyl, C 1-10 Halogenated hydrocarbon group, three fluorine acetyl, halogenated phenyl, halogenated benzyl, nitro phenyl or any kind of groups substituted form. Among them, the substituent atom or the substituent is preferably a fluorine atom, an alkoxy group or a nitro group.
LG 2 More preferably, it is any of tert-butyl, trityl, phenyl, benzyl, methylbenzyl, tert-butylthio, benzylthio, 2-pyridylthio, 2-pyridylcarbonyl, tert-butyloxycarbonyl, phenoxycarbonyl, benzyloxycarbonyl, tert-butyloxythiocarbonyl, phenoxythiocarbonyl, benzyloxythiocarbonyl, tert-butylthiocarbonyl, phenylthiocarbonyl, benzylthiocarbonyl, trifluoroacetyl and the like.
LG 2 More preferably, it is any of tert-butyl, trityl, phenyl, benzyl, methylbenzyl, tert-butylthio, benzylthio, 2-pyridylthio and the like.
LG 2 Most preferred is methyl, ethyl, allyl or benzyl.
Wherein, X 6 Is a terminal group attached to the oxygen atom of the ester group and is selected from a hydroxyl protecting group or the group LG 4 . Wherein LG is 4 The definitions of (a) and (b) are consistent with the above.
In this case, Q is not particularly limited as long as it contributes to the induction of unsaturated bond electrons and the conjugation effect.
When Q is on the ring, it may be one or more. When a plurality of such structures are provided, the same structure may be provided, or a combination of two or more different structures may be provided.
Q may be an atom or a substituent.
When atomic, Q is selected from a hydrogen atom or a halogen atom, preferably a hydrogen atom or a fluorine atom.
When a substituent, Q is selected from the group consisting of, but not limited to, all combinations of substituents listed in the term part. May or may not contain carbon atoms. In the case where no carbon atom is contained, for example, a nitro group may be mentioned. When carbon atoms are contained, the number of carbon atoms is not particularly limited, but 1 to 20 carbon atoms are preferable, and 1 to 10 carbon atoms are more preferable.
When a substituent, the structure of Q is not particularly limited, and includes, but is not limited to, a linear structure, a branched structure containing a pendant group, or a cyclic structure. Among them, the cyclic structure is not particularly limited, and the above-mentioned aliphatic ring, aromatic ring, sugar ring, and condensed ring are preferable.
Q may be selected from any one atom or group of a hydrogen atom, a halogen atom, a non-carbon containing substituent, a hydrocarbyl group, a heterohydrocarbyl group, a substituted hydrocarbyl group or a substituted heterohydrocarbyl group.
Q is preferably a hydrogen atom, a halogen atom, a nitro group-containing substituent, an acyl group-containing substituent, or C 1-20 Haloalkyl, C 1-20 Alkyl radical, C 2-20 Alkenyl radical, C 3-20 Open-chain alkylene radical, C 3-20 Cycloalkyl, aryl, arylalkyl, C 1-20 Heteroalkyl, heteroaryl, heteroaralkyl, C 1-20 Alkoxy, aryloxy, aralkyloxy, C 1-20 Heteroalkyloxy, heteroaryloxy, heteroarylhydrocarbyloxy, C 1-20 Alkylthio, arylthio, aralkylthio, C 1-20 Any one atom or group, or substituted version of any one group, of heteroalkylthio, heteroarylthio, heteroarylalkylthio, and the like. Wherein the substituted heteroatom or substituent in Q is not particularly limited, including but not limited to the term moietyAny one of the substituted hetero atoms or any one of the substituents of (1) is selected from any one of a halogen atom, a hydrocarbon group substituent and a heteroatom-containing substituent.
Q is more preferably a hydrogen atom, a halogen atom, a nitro group-containing substituent, an acyl group, an ester group-containing substituent at the terminal group, a thioester group-containing substituent at the terminal group, an amide bond-containing substituent at the terminal group, C 1-20 Haloalkyl, C 2-20 Alkenyl radical, C 3-20 Open-chain alkylene radical, C 3-20 Cycloalkyl, aryl, arylalkyl, C 1-20 Heteroalkyl, heteroaryl, heteroaralkyl, C 1-20 Alkoxy, aryloxy, C 1-20 Heteroalkyloxy, heteroaryloxy, heteroarylhydrocarbyloxy, C 1-20 Alkylthio, arylthio, aralkylthio, C 1-20 Heteroalkylthio, heteroarylthio, heteroarylalkylthio, and the like, or a substituted version of any group. Wherein, the acyl group is not particularly limited, including but not limited to any acyl type of the term moiety. By way of example, the acyl group in Q may be selected from carbonyl, sulfonyl, sulfinyl, phosphoryl, phosphorylidene, hypophosphoryl, nitroxyl, nitrosyl, thiocarbonyl, imidoyl, thiophosphoryl, dithiophosphoryl, thiophosphoryl, thiophosphorylidene, dithiophosphorylidene, thiophosphorylidene, thiophosphonyl, dithiophosphono, thiophosphinyl, and the like. Any of acyl groups such as a carbonyl group, a thiocarbonyl group, a sulfonyl group, and a sulfinyl group is preferable. More preferably, the acyl group is a carbonyl group, thiocarbonyl group, sulfonyl group or sulfinyl group.
Q is more preferably a hydrogen atom, a halogen atom, a nitro group-containing substituent, or C 1-20 Carbonyl group, C 1-20 Alkylthio carbonyl of C 1-20 Sulfonyl radical, C 1-20 Alkyloxycarbonyl, C 1-20 Alkylthio carbonyl group, C 1-20 Alkylaminocarbonyl radical, C 1-20 Alkyloxythiocarbonyl radical, C 1-20 Alkylthio thiocarbonyl radical, C 1-20 Alkylaminothiocarbonyl, C 1-20 Alkyloxysulfonyl, C 1-20 Alkyloxysulfinyl, arylthioCarbonyl, aryloxycarbonyl, arylthiocarbonyl, arylaminocarbonyl, aryloxythocarbonyl, arylthiothiocarbonyl, arylaminothiocarbonyl, aryloxysulfonyl, aryloxysulfinyl, aralkylthiocarbonyl, aralkyloxycarbonyl, aralkylthiocarbonyl, aralkylaminocarbonyl, aralkyloxythiocarbonyl, aralkylaminothiocarbonyl, aralkyloxysulfonyl, aralkyloxysulfinyl, C 1-20 Alkyl radical, C 2-20 Alkenyl radical, C 3-20 Open-chain alkenyl, C 3-20 Cycloalkyl, aryl, arylalkyl, C 1-20 Heteroalkyl, heteroaryl, heteroaralkyl, C 1-20 Alkoxy, aryloxy, aralkyloxy, C 1-20 Heteroalkyloxy, heteroaryloxy, heteroarylhydrocarbyloxy, C 1-20 Alkylthio, arylthio, aralkylthio, C 1-20 Heteroalkylthio, heteroarylthio, heteroarylalkylthio, C 1-20 Haloalkyl, and the like, or a substituted version of either atom or group.
Q is more preferably a hydrogen atom, a halogen atom, a nitro group-containing substituent, or C 1-10 Carbonyl group, C 1-10 Alkylthio carbonyl group, C 1-10 Sulfonyl, C 1-10 Alkyloxycarbonyl, C 1-10 Alkylthio carbonyl group, C 1-10 Alkylaminocarbonyl radical, C 1-10 Alkyloxythiocarbonyl radical, C 1-10 Alkylthio thiocarbonyl radical, C 1-10 Alkylamino thiocarbonyl radical, C 1-10 Alkyloxysulfonyl, C 1-10 Alkyloxysulfinyl, arylthiocarbonyl, aryloxycarbonyl, arylthiocarbonyl, arylaminocarbonyl, aryloxysulfonyl, aryloxysulfinyl, aralkylthiocarbonyl, aralkyloxycarbonyl, aralkylthiocarbonyl, aralkylaminocarbonyl, aralkyloxythiocarbonyl, aralkylthiocarbonyl, aralkylthiothiocarbonyl, aralkyloxysulfonyl, aralkyloxysulfinyl, C 1-20 Alkyl radical, C 2-10 Alkenyl radical、C 3-10 Open-chain alkenyl, C 3-10 Cycloalkyl, aryl, arylalkyl, C 1-10 Heteroalkyl, heteroaryl, heteroaralkyl, C 1-10 Alkoxy, aryloxy, aralkyloxy, C 1-10 Heteroalkyloxy, heteroaryloxy, heteroarylhydrocarbyloxy, C 1-10 Alkylthio, arylthio, aralkylthio, C 1-10 Heteroalkylthio, heteroarylthio, heteroarylalkylthio, C 1-10 Haloalkyl, and the like, or a substituted version of either atom or group.
Specifically, Q may be selected from the group consisting of a hydrogen atom, a fluorine atom, a chlorine atom, a bromine atom, an iodine atom, a nitro group, a nitrophenyl group, an acetyl group, a benzoyl group, a p-toluenesulfonate group, a methanesulfonate group, a methoxycarbonyl group, an ethoxycarbonyl group, a tert-butyloxycarbonyl group, a phenoxycarbonyl group, a benzyloxycarbonyl group, a methylthioacyl group, an ethylthioacyl group, a tert-butylthiocarbonyl group, a phenylthiocarbonyl group, a benzylthiocarbonyl group, an ethylaminoacyl group, a tert-butylaminocarbonyl group, a phenylaminocarbonyl group, a benzylaminocarbonyl group, a methoxythiocarbonyl group, an ethoxythiocarbonyl group, a tert-butyloxycarbonyl group, a phenoxythiocarbonyl group, a benzyloxythiocarbonyl group, a methylthiothiocarbonyl group, an ethylthiothiocarbonyl group, a tert-butylthiocarbonyl group, a phenylthiocarbonyl group, a benzylthiocarbonyl group, an ethylaminoacyl group, a tert-butylaminothiocarbonyl group, a, Phenylaminothiocarbonyl, benzylaminothiocarbonyl, methyl, ethyl, n-propyl, isopropyl, butyl, pentyl, hexyl, heptyl, 2-ethylhexyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, eicosyl, vinyl, propenyl, allyl, propynyl, propargyl, cyclopropyl, cyclopropenyl, phenyl, benzyl, butylphenyl, p-methylphenyl, methoxy, ethoxy, phenoxy, benzyloxy, methylthio, ethylthio, phenylthio, benzylthio, C 1-20 Haloalkyl, and the like, or substituted versions of either group. Wherein, butyl includes but is not limited to n-butyl and tert-butyl. Octyl includes but is not limited to nOctyl and 2-ethylhexyl. Among them, the substituent atom or the substituent is selected from any one of a halogen atom, a hydrocarbon substituent and a heteroatom-containing substituent, and is preferably a halogen atom, an alkoxy group, an alkenyl group, an aryl group or a nitro group.
Q is preferably a hydrogen atom, a fluorine atom, a chlorine atom, a bromine atom, an iodine atom, a nitro group, a nitrophenyl group, an acetyl group, a benzoyl group, a p-toluenesulfonyl group, a methanesulfonic group, a methoxyacyl group, an ethoxyacyl group, a tert-butyloxycarbonyl group, a phenoxycarbonyl group, a benzyloxycarbonyl group, a methylthioacyl group, an ethylthioacyl group, a tert-butylthiocarbonyl group, a phenylthiocarbonyl group, a benzylthiocarbonyl group, an ethylaminoacyl group, a tert-butylaminocarbonyl group, a phenylaminocarbonyl group, a benzylaminocarbonyl group, a methyl group, an ethyl group, a n-propyl group, an isopropyl group, a butyl group, a pentyl group, a hexyl group, a heptyl group, an octyl group, a nonyl group, a decyl group, a vinyl group, a propenyl group, an allyl group, a propynyl group, a propargyl group, a cyclopropyl group, a cyclopropenyl group, a phenyl group, a benzyl group, a butylphenyl group, a p-methylphenyl group, a methoxy group, an ethoxy group, a phenoxy group, a benzyloxy group, a methylthio group, an ethylthio group, a phenylthio group, Any atom or group, or substituted version of any group, of benzylthio, trifluoromethyl, 2,2, 2-trifluoroethyl, and the like. Among them, the substituent atom or the substituent is preferably a fluorine atom, an alkoxy group, an alkenyl group, an aryl group or a nitro group.
Q is more preferably any one atom or group selected from a hydrogen atom, a fluorine atom, a methyl group, a trifluoromethyl group, a methoxy group, a methyloxycarbonyl group, a p-toluenesulfonyl group, a methanesulfonyl group and the like.
Q is more preferably any one atom or group selected from a hydrogen atom, a fluorine atom, a methyl group, a trifluoromethyl group, a methoxy group, a methyloxycarbonyl group and the like.
Wherein Q 3 Is an H atom or a group that contributes to the induction, conjugation effect of unsaturated bond electrons;
Q 3 selected from the group consisting of, but not limited to, all of the substituent atoms and combinations of substituents listed in the term part, as long as they contribute to the induction, conjugation effect of the unsaturated bond electrons.
Q 3 May or may not contain carbon atoms. When no carbon atom is contained, for example, it may beA nitro group. When a carbon atom is contained, the number of carbon atoms is not particularly limited, but is preferably 1 to 20 carbon atoms, more preferably 1 to 10 carbon atoms.
Q 3 The structure of (a) is not particularly limited and includes, but is not limited to, a linear structure, a branched structure containing a side group, or a cyclic structure. Among them, the cyclic structure is not particularly limited, and the above-mentioned aliphatic ring, aromatic ring, sugar ring, and condensed ring are preferable.
Q 3 Can be selected from any atom or group of hydrogen atom, halogen atom, substituent without carbon, alkyl, heteroalkyl, substituted alkyl or substituted heteroalkyl. Wherein Q is 3 The substituted heteroatom or substituent in (1) is not particularly limited, including but not limited to any substituted heteroatom or any substituent of the term moiety selected from any of halogen atoms, hydrocarbyl substituents, heteroatom-containing substituents.
Q 3 More preferably a hydrogen atom, a halogen atom, C 1-20 Alkyl radical, C 2-20 Alkenyl radical, C 3-20 Open-chain alkenyl, C 3-20 Cycloalkyl, aryl, arylalkyl, C 1-20 Heteroalkyl, heteroaryl, heteroaralkyl, C 1-20 Alkoxy, aryloxy, aralkyloxy, C 1-20 Heteroalkyloxy, heteroaryloxy, heteroarylhydrocarbyloxy, C 1-20 Heteroalkylthio, heteroarylthio, heteroarylalkylthio, C 1-20 Haloalkyl, and the like, or substituted versions of either group.
Q 3 More preferably a hydrogen atom, a halogen atom, C 1-10 Haloalkyl, C 1-10 Alkyl radical, C 2-10 Alkenyl radical, C 3-10 Open-chain alkenyl, C 3-10 Cycloalkyl, aryl, arylalkyl, C 1-10 Heteroalkyl, heteroaryl, heteroaralkyl, C 1-10 Alkoxy, aryloxy, aralkyloxy, C 1-10 Any atom or group, or substituted version of any group, of heteroalkyloxy, heteroaryloxy, heteroarylhydrocarbyloxy, and the like.
Specifically, Q 3 Can be selected from hydrogen atom, fluorine atom, chlorine atom, bromine atom, iodine atom Methyl, ethyl, n-propyl, isopropyl, butyl, pentyl, hexyl, heptyl, 2-ethylhexyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, eicosyl, vinyl, propenyl, allyl, propynyl, propargyl, cyclopropyl, cyclopropenyl, phenyl, benzyl, butylphenyl, p-methylphenyl, nitrophenyl, p-methoxyphenyl, azaphenyl, methoxy, ethoxy, phenoxy, benzyloxy, methylthio, ethylthio, phenylthio, benzylthio, C 1-20 Haloalkyl, and the like, or substituted versions of either group. Wherein, butyl includes but is not limited to n-butyl and t-butyl. Octyl includes, but is not limited to, n-octyl, 2-ethylhexyl. Among them, the substituent atom or the substituent is selected from any one of a halogen atom, a hydrocarbon substituent and a heteroatom-containing substituent, and is preferably a halogen atom, an alkoxy group, an alkenyl group or a nitro group.
Q 3 Preferably any one atom or group selected from the group consisting of a hydrogen atom, a fluorine atom, a chlorine atom, a bromine atom, an iodine atom, a methyl group, an ethyl group, an n-propyl group, an isopropyl group, a butyl group, a pentyl group, a hexyl group, a heptyl group, an octyl group, a nonyl group, a decyl group, a vinyl group, a propenyl group, an allyl group, a propynyl group, a propargyl group, a cyclopropyl group, a cyclopropenyl group, a phenyl group, a benzyl group, a butylphenyl group, a p-methylphenyl group, a p-nitrophenyl group, an o-nitrophenyl group, a p-methoxyphenyl group, an azaphenyl group, a methoxy group, an ethoxy group, a phenoxy group, a benzyloxy group, a methylthio group, an ethylthio group, a phenylthio group, a benzylthio group, a trifluoromethyl group, a 2,2,2, 2-trifluoroethyl group and the like, or a substituted form of any one of the groups. Among them, the substituent atom or the substituent is preferably a fluorine atom, an alkoxy group, an alkenyl group or a nitro group. The azaphenyl group is preferably pyridyl, pyrimidine, pyrazine, 1,3, 5-triazine.
Q 3 More preferably any one atom or group of a hydrogen atom, a methyl group, a trifluoromethyl group, a phenyl group, a p-nitrophenyl group, an o-nitrophenyl group, a pyridyl group or a substituted form thereof, a diazophenyl group or a substituted form thereof, a triazophenyl group or a substituted form thereof, and the like.
Q 3 More preferably a hydrogen atom, methyl group, phenyl group, pyridyl group, diazophenyl group, triazophenyl group.
Q 3 More preferably a hydrogen atom, a methyl group, a phenyl group or a pyridyl group.
Q 3 Most preferably a hydrogen atom, a phenyl group or a pyridyl group.
Wherein Q is 6 Is a hydrogen atom or a methyl group. Q 7 Is hydrogen atom, methyl, phenyl or substituted phenyl. Such as p-methoxyphenyl. In the same molecule, Q 6 And Q 7 May be the same or different.
Wherein Q is 8 The substituent atom or substituent on the imidazolyl group is not particularly limited, and is preferably selected from the group consisting of an H atom, a methyl group, an ethyl group, a propyl group, a butyl group and a phenyl group. When Q is 8 May be one or more. When the number is more than 1, the structures may be the same, or a combination of two or more different structures may be used.
Wherein Q 11 Is a substituent on the nitrogen atom of tetrazole, preferably phenyl, substituted phenyl or aza phenyl.
Wherein PG 2 Is a thiol protecting group, the protected thiol group being denoted as SPG 2 。
Wherein PG 3 Being an alkynyl protecting group, the protected alkynyl group is represented by C ≡ CPG 3 。
Wherein PG 4 Is a hydroxy protecting group, the protected hydroxy group being represented by OPG 4 。
Wherein PG 5 Is an amino protecting group, the protected amino group being represented by NPG 5 。
PG 2 、SPG 2 、PG 3 、PG 4 、OPG、PG 5 、NPG 5 Including but not limited to the structures described and exemplified in documents CN104877127A, CN104530413A, CN104530415A, CN104530417A and the respective cited documents. Take CN104530417A as an example, corresponding to section [0520]～[0530]. In general terms, the use of a single,
the SPG 2 Preferred are thioether, disulfide, silyl sulfide, thioester, and the like. In particular, SPG 2 Preference is given toT-butyl sulfide, trityl sulfide, substituted trityl sulfide, t-butyl dimethyl silyl sulfide, triisopropyl silyl sulfide, benzyl sulfide, substituted benzyl sulfide, p-nitrobenzyl sulfide, o-nitrobenzyl sulfide, acetyl thioester, benzoyl thioester, trifluoroacetyl thioester, t-butyl disulfide, substituted phenyl disulfide, 2-pyridine disulfide, or the like.
The PG 3 Preferred silicon groups include, but are not limited to, the following structures: trimethylsilyl group, triethylsilyl group, t-butyldimethylsilyl group, dimethyl (1,1, 2-trimethylpropyl) silyl group, dimethyl [1, 1-dimethyl-3- (tetrahydrofuran-2H-2-oxy) propyl group ]Silicon group, biphenyl dimethyl silicon group, triisopropyl silicon group, biphenyl diisopropyl silicon group, tert-butyl diphenyl silicon group, 2- (2-hydroxy) propyl group and the like.
The PG 4 It may be a protecting group for alcoholic hydroxyl group or phenolic hydroxyl group. OPG 4 Preferred are ether, silyl ether, ester, carbonate, sulfonate, and the like. In particular, OPG 4 Preferably methyl ether, 1-ethoxy ethyl ether, tert-butyl ether, allyl ether, benzyl ether, p-methoxy benzyl ether, o-nitrobenzyl ether, p-nitrobenzyl ether, 2-trifluoromethyl benzyl ether, methoxy methyl ether, 2-methoxyethoxymethyl ether, benzyloxy methyl ether, p-methoxybenzyloxymethyl ether, methylthio methyl ether, tetrahydropyranyl ether, trimethylsilyl ether, triethylsilyl ether, triisopropylsilyl ether, tert-butyldimethylsilyl ether, acetate, chloroacetate, trifluoroacetate, carbonate, etc. Among the ether protecting structures, 1-ethoxyethyl ether, benzyl ether, p-methoxybenzyl ether, o-nitrobenzyl ether, p-nitrobenzyl ether, 2-trifluoromethylbenzyl ether, vinyl ethyl ether, benzyloxymethyl ether, p-methoxybenzyloxymethyl ether, tetrahydropyranyl ether are preferable.
The PG 5 May be a protecting group for primary amine, secondary amine, hydrazine, etc. NPG 5 Preferred are structures such as carbamate, amide, imide, N-alkylamine, N-arylamine, imine, enamine, imidazole, pyrrole, indole, and the like. In particular, NPG 5 Preferably formamide, acetamide and trifluoroethylAmide, carbamic acid tert-butyl ester, carbamic acid 2-iodoethyl ester, carbamic acid benzyl ester, carbamic acid 9-fluorenylmethyl ester, carbamic acid 2-trimethylsilylethyl ester, carbamic acid 2-methylsulfonylethyl ester, carbamic acid 2- (p-toluenesulfonyl) ethyl ester, phthalimide, diphenylmethyleneamine, 1,3, 5-dioxaazacyclohexyl, methylamino, triphenylmethylamino, tert-butylamino, allylamino, benzylamino, 4-methoxybenzylamino, benzylimine, and the like.
Wherein PG 6 Is a bishydroxy protecting group, and PG 6 And two oxygen atoms form an acetal structure of a five-membered ring or a six-membered ring. PG (Picture experts group) 6 Selected from methylene or substituted methylene. The PG 6 The substituent(s) of (a) is a hydrocarbyl substituent or a heteroatom-containing substituent, including, but not limited to, the following: methylene, 1-methylmethylene, 1-dimethylmethylene, 1-cyclopentylene, 1-cyclohexylene, 1-phenylmethylene, 3, 4-dimethylphenylmethylene, and the like.
1.6.4. Containing functional groups- (Z) 1 ) q1 -R 01 Examples of (2)
By way of example, - (Z) 2 ) q -(Z 1 ) q1 -R 01 Including but not limited to the structures described and exemplified in documents CN104877127A, CN104877127A, CN104530413A, CN104530415A, CN104530417A and the respective cited documents. Including but not limited to CN104530417A paragraph [0531]～[0592]CN104877127A paragraph [0458]～[0505]The structures listed.
1.7. Description of the groups which can be stabilized and degraded
The linker STAG or the degradable linker DEGG which can be stably present in the present invention may be present in the above-mentioned L U0 、L 2 、 L 0 (g＝1)、Z 1 、Z 2 Any of the divalent linking groups, or a divalent linking group present between any of the divalent linking groups and an adjacent heteroatom group, may also be present in any of the multivalent groups U 0 G, or any divalent linking group formed by a polyvalent group and an adjacent group.
The nitrogen-branched nine-arm polyethylene glycol derivative can be stabilizedPresent or degradable. When degradable, the number of degradable sites in the same molecule may be 1 or more. With respect to the degradable site: (1) can be positioned at U 0 、L 2 、L 0 、G、(Z 2 ) q -(Z 1 ) q1 Any one of the degradable groups; (2) but not for Z, at the junction of any of the above groups with an adjacent group 1 -R 01 The degradability of the attachment site of (a) is defined. In the first case, the degradable group may contain a degradable divalent linking group DEGG such as an ester group, a carbonate group, etc. For the second case, then, one can choose to be on O-L 0 、L 0 -G、G-Z 2 、Z 2 -Z 1 Degradation of either attachment site can occur.
According to the number of degradable sites and the position difference of the degradable sites in the nitrogen-branched nine-arm polyethylene glycol derivative, the stability of the polymer and the releaseability of the modified drug are greatly influenced. When degradation can occur between the functional group at the end of the nine polyethylene glycol chains and the polyethylene glycol chain, i.e. - (Z) 2 ) q -(Z 1 ) q1 -the position of the drug molecule and the polyethylene glycol structure are separated, so that the active site of the drug molecule is exposed to the maximum extent, and the drug molecule can approach the unmodified state to the maximum extent.
Several degradation modes are typical as follows: (a) when g is 0 or 1, when only in L 2 When the connection position of the PEG is degraded, the PEG can be degraded into nine PEG single-chain structures; (b) when g is 0 or 1, when g is only in- (Z) 2 ) q -(Z 1 ) q1 -a degradation of the position containing its connection group with the adjacent group on the PEG side into a nitrogen branched nine-arm PEG structure with an independent functional group residue; (c) when g is 1, and is only in L 0 Position (including L) 0 Internal, O-L 0 Connection, L 0 -G-attachment) to a nitrogen branched nine-arm PEG structure and a cluster of multiple functional groups attached through G; (d) when G ═ 1, and degradation occurs only in G, degradation to nitrogen branched nine-arm PEG, individual functional group residues, or residues of multiple functional clusters; (e) when g is 0 or 1, only in U 0 When the connecting position is degraded, the structure can be degraded into nine PEG single-chain structures or three branched three-arm polyethylene glycol structures.
1 or more than 1 degradation mode is allowed to exist in the nitrogen branched nine-arm polyethylene glycol derivative. When more than one degradation mode exists, gradient degradation can occur, and the degradation kinetic process of the modified product can be controlled more flexibly; for PEG modified drugs, the control of pharmacokinetics in vivo is more flexible and fine, and the requirement of complex treatment effect can be met.
1.7.1. Stag, a bivalent connecting group stably existing in the present invention
The stable presence condition of STAG is not particularly limited, and it may be stable under any condition including, but not limited to, light, heat, low temperature, enzyme, redox, acidic, alkaline condition, physiological condition, in vitro simulated environment, and the like, and preferably, it may be stable under any condition of light, heat, enzyme, redox, acidic, alkaline, and the like.
STAG types are not particularly limited and include, but are not limited to, alkylene groups, divalent heteroalkyl groups, double bonds, triple bonds, divalent dienyl groups, divalent cycloalkyl groups, divalent cycloalkenyl groups, divalent cycloalkenylene groups, divalent cycloalkynylene groups, aromatic groups, alicyclic groups, heteroheterocyclic groups, heteroaromatic groups, substituted alkylene groups, substituted divalent heteroalkyl groups, substituted double bonds, substituted dienyl groups, substituted divalent cycloalkyl groups, substituted divalent cycloalkenyl groups, substituted divalent cycloalkenylene groups, substituted divalent cycloalkynylene groups, substituted aromatic groups, substituted alicyclic groups, substituted heteroaromatic groups, ether linkages, thioether linkages, urea linkages, thiourea linkages, carbamate groups, thiocarbamate groups, divalent silicon groups free of active hydrogen, divalent linking groups containing a boron atom, Secondary amino, tertiary amino, carbonyl, thiocarbonyl, amido, thioamido, sulfamide, enamine, triazolyl, 4, 5-dihydro isoxazolyl, any divalent linking group in skeleton of amino acid and its derivative, and stable divalent linking group composed of any two or more groups.
In particular, STAGs include, but are not limited to, the structures described and exemplified in documents CN104530413A, CN104530415A, CN 104530417A. For example, CN104530417A corresponds to the segments [0627] to [0704 ]. The manner in which two or more species of bivalent linking groups that can be stably present are combined to form STAG is not particularly limited. Including but not limited to segment [704] of CN 104530417A.
As a combination of any two or more of the structures, for example, -CH 2 O-、-OCH 2 -、-CH 2 CH 2 O-、 -OCH 2 CH 2 -、-OCH 2 CH 2 O-、-(CH 2 ) 3 O-、-O(CH 2 ) 3 -、-(CH 2 ) 3 O-、-O(CH 2 ) 3 -and the like. As examples of L 0 An oligopeptide or polypeptide or the like formed end-to-end of the N-segment and C-terminus, which may comprise a plurality of amino acids, which may be the same or different, but does not include a polypeptide fragment that is degradable by in vivo biological enzymes. Furthermore, L 0 In one aspect, the composition may further comprise (L) 7 O) nj -、 -(OL 7 ) nj -、-(R 29 O) nj -、-(OR 29 ) nj -、-(CH 2 CH 2 O) nj -、-(OCH 2 CH 2 ) nj -and the like. Wherein L is 7 、 R 29 The definitions of (a) and (b) are consistent with the above. Wherein the integer nj is the number of the repeating units with a monodisperse structure, and is selected from 2-20, preferably 2-10. CORE of the invention 8 (O-) 8 In which no other than-CH is included 2 CH 2 A heteroatom-containing repeating unit other than O-.
1.7.2. Degradable bivalent connecting group DEGG in the invention
The DEGG is degradable under any conditions including, but not limited to, light, heat, low temperature, enzyme, redox, acidic, basic, physiological conditions, in vitro simulated environment, and the like, preferably under any conditions of light, heat, enzyme, redox, acidic, basic, and the like.
The divalent linking group formed by any combination of DEGG and any STAG remains a degradable linking group. For degradable divalent linking groups containing aromatic rings, the aromatic rings can also be combined with the degradable divalent linking groups.
The type of DEGG is not particularly limited and includes, but is not limited to, those containing disulfide bonds, vinyl ether bonds, ester groups, thioester groups, dithioester groups, carbonate groups, thiocarbonate groups, dithiocarbonate groups, trithiocarbonate groups, carbamate groups, thiocarbamate groups, dithiocarbamate groups, acetal groups, cyclic acetal groups, mercaptal groups, azaacetal groups, azathiolacetal groups, dithioacetal groups, hemiacetal groups, thiohemiacetal groups, azahemiacetal groups, ketal groups, mercaptide groups, azaketal groups, azathioketal groups, imine bonds, hydrazone bonds, acylhydrazone bonds, oxime bonds, sulfoximine ether groups, semicarbazone bonds, thiosemicarbazone bonds, hydrazine groups, hydrazide groups, thiocarbohydrazide groups, thioazocarbohydrazide groups, Hydrazinoformate groups, hydrazinothiocarbamate groups, carbazoyl groups, thiocarbazoic groups, isothiourea groups, allophanate groups, thioallophanate groups, guanidino groups, amidino groups, aminoguanidino groups, amidino groups, imido groups, thioester groups, sulfonate groups, sulfinate groups, sulfonylhydrazine groups, sulfonylurea groups, maleimide groups, orthoester groups, phosphate groups, phosphite groups, hypophosphite groups, phosphonate groups, phosphosilane groups, silane groups, carbonamide groups, thioamide groups, sulfonamide groups, polyamide groups, phosphoramide groups, phosphoramidite groups, pyrophosphoamide groups, cyclic phosphoramide groups, isocyclophosphamide groups, thiophosphoramide groups, aconityl groups, polypeptide fragments, backbones of nucleotides and derivatives thereof, divalent linking groups of any of deoxynucleotides and derivative backbones thereof, Any combination of two or more divalent linking groups.
The urethane group, thiocarbamate group, carbonamide group, phosphoramide group, etc. herein may be used as a linker which may exist stably, or may be used as a degradable linker. Depending on the environmental characteristics of its use.
In particular, DEGG includes, but is not limited to, the structures described and exemplified in documents CN104530413A, CN104530415A, CN 104530417A. For example, CN104530417A corresponds to segments [705] to [0725 ].
1.7.3. Degradable polyvalent radical
The degradable trivalent or tetravalent or higher group needs to contain at least one degradable divalent linking group DEGG.
For degradable trivalent groups, there are included, but not limited to, groups of stable trivalent groups containing trivalent atomic nucleus structures with degradable divalent linkers, groups of trivalent aromatic rings with degradable divalent linkers, combinations of degradable trivalent ring structures with divalent linkers that may be present stably, combinations of degradable trivalent ring structures with degradable divalent linkers, trivalent forms of any of the above degradable divalent linkers. Wherein a degradable trivalent ring structure refers to a trivalent ring structure that is degradable into at least two separate segments. The structure can be a trivalent closed ring structure formed by connecting 2 or more degradable groups in series. For example, cyclic peptides have a cyclic structure in which 2 or more ester bonds are connected in series. Including but not limited to the structures described and exemplified in documents CN104530413A, CN104530415A, CN 104530417A. For example, CN104530417A corresponds to segments [726] to [0731 ].
1.8. Examples of terminal branched structures G
The structure of G is not particularly limited, and each independently includes, but is not limited to, branched, cyclic structure-containing, comb-like, dendritic, hyperbranched, and the like. G may be degradable or may exist stably.
L 0 The divalent linking group, which is used to link the PEG segment to the terminal branched structure G, may or may not be present. L is 0 Can exist stably or can be degraded. L is 0 May be selected from any of the foregoing STAG or DEGG.
The terminal branching groups G of the nitrogen-branched nine-arm polyethylene glycol derivative have the same structure type, such as a three-branched structure, a four-branched structure, a comb-shaped structure, a tree-shaped structure, a hyperbranched structure or a cyclic structure. In the case of the same structure type, the structure allowing nine PEG chain ends is not completely identical. For example, for a comb structure, differences in valence states due to the non-uniform number of repeating units are allowed; for hyperbranched structures, the number of branching units is not required to be strictly uniform, but rather allows the branching units to be randomly linked. Therefore, in the same molecule, when the end of the PEG chain is in a comb-like or super-branched structure, the k at the end may be different. For tree and ring structures, the structures are required to be completely consistent, and the corresponding k is also completely equal.
In the case where the nitrogen-branched nonaarm polyethylene glycol derivative G ═ 1 in the present invention, G is selected from the group including, but not limited to, any of the above-mentioned k +1(k ═ 2 to 250) valent groups. Preferred structures of G include, but are not limited to, the structures described and exemplified in documents CN104530413A, CN104530415A, CN 104530417A. For example, CN104530417A corresponds to segments [0824] to [0825 ].
When the terminal reaction site k is 2, G is a trivalent group. L is 0 -G preferably comprises a structure selected from the group consisting of:
Figure BDA0002963906360000431
Figure BDA0002963906360000432
Figure BDA0002963906360000441
Figure BDA0002963906360000442
and the like; wherein, the asterisk marks in the structure, which indicates that the asterisk end points to the polyethylene glycol unit; the G can also be a trivalent framework structure of amino acid or a derivative thereof; wherein the amino acid is L Type-or D -type; the amino acid or the derivative thereof is derived from any one of the following: serine, threonine, cysteine, leucine, hydroxyproline, aspartic acid, glutamic acid, asparagine, glutamine, lysine, arginine, citrulline, histidine, tryptophan.
When the terminal reaction site k is 3, G is a tetravalent group, including but not limited to the above group G 4 In (1)A tetravalent group; tetravalent G preferably contains the atom CM 4 And unsaturated bond CB 4 Ring structure CC 4 Either tetravalent core structure, or contains two trivalent core structures. L is 0 -G further preferably comprises any of the following structures:
Figure BDA0002963906360000451
Figure BDA0002963906360000461
Figure BDA0002963906360000462
etc.; wherein the asterisks in the structure indicate that the asterisk ends point towards the polyethylene glycol unit.
When k.gtoreq.3 is the terminal reaction site, i.e., G has a valence of 4 or more, G having a valence of k +1 includes, but is not limited to, the above-mentioned group G k+1 The k + 1-valent group in (1). The G with the valence of k +1 can contain 1 nuclear structure with the valence of k +1, or is formed by directly connecting and combining 2-k-1 low-valence groups with the valence of 3-k or is formed by 1 or more than 1 divalent spacer groups L 10 Indirectly combined together. The 3 to k-valent lower groups may be the same or different, and the valences thereof may be the same or different. For example: two different trivalent groups are combined
Figure BDA0002963906360000463
For a k +1 valent core structure, when k.gtoreq.4, and when a k +1 valent core structure is contained, the k +1 valent core structure is preferably a cyclic structure. When containing two or more L 10 When L is 10 May be the same as or different from each other. L is a radical of an alcohol 10 The definition of (c) is consistent with the above.
The direct or indirect combination of K +1(k is more than or equal to 4) G, the combination mode includes but is not limited to comb combination mode, tree combination mode, branched combination mode, hyperbranched combination mode, cyclic combination mode, etc. For example, in the case of a comb-like, tree-like or hyperbranched group in which a plurality of low-valent groups are combined, the plurality of low-valent groups may be the same as or different from each other, and are preferably combined from the same low-valent group.
Wherein, the tree-like combination structure formed by the tree-like combination mode is DENR (U) denr NONE, d) or DENR (U) denr ,L 10 D) represents U denr Represents a polyvalent radical repeating unit, NONE represents a direct linkage of polyvalent repeating units, L 10 Denotes a polyvalent repeating unit via a divalent linking group L 10 Indirectly connected, d represents the generation of a tree combination mode, d is preferably 2-6 generations, more preferably 2-5 generations, and most preferably 2, 3 or 4 generations. Among them, the basic unit of the polyvalent G constituting the tree-like composite structure is preferably trivalent G or tetravalent G.
Examples of the tree-like composite structure are
Figure BDA0002963906360000464
Figure BDA0002963906360000465
Figure BDA0002963906360000471
Figure BDA0002963906360000472
And the like. Wherein ng represents an algebra of a tree combination mode.
Among them, the basic unit of the polyvalent G constituting the branched or hyperbranched composite structure is preferably trivalent G or tetravalent G. The preferred basic units include, but are not limited to, the tree combination method described above, and further include
Figure BDA0002963906360000473
And the like. The branched or hyperbranched composite structure differs from the above-described dendritic composite structure in that it is a mixed combination of multivalent G and its lower valent form. Lower forms of said polyvalent G, for example
Figure BDA0002963906360000474
Is selected from
Figure BDA0002963906360000475
Wherein the asterisks within the structure indicate that the asterisk end points to the polyethylene glycol unit.
Among them, the basic unit of the polyvalent G constituting the comb-like composite structure is preferably trivalent G, tetravalent G or pentavalent G. Preferred are the repeating units described in the stages [824] to [0825] of CN104530417A, and the stages [1130] and [1143] of CN 104530413A. The basic unit of the polyvalent G constituting the comb-like composite structure includes, but is not limited to, polyglycerol, polypentaerythritol, substituted propylene oxide, a group of substituted propylene oxide and carbon dioxide, acrylate and its derivatives, methacrylate and its derivatives, acetal structure-containing basic units (e.g., (1 → 6) β -D glucopyranoside), hydroxyl or thio group-containing amino acids and their derivatives, acidic amino acids and their derivatives, basic amino acids and their derivatives, and the like. G may be acetalized dextran formed by connecting D-glucopyranose units end to end via any one of the bond forms of β -1,6 glycosidic bond, α -1,6 glycosidic bond, β -1,4 glycosidic bond, α -1,4 glycosidic bond, β -1,3 glycosidic bond and α -1,3 glycosidic bond, or an oxidized form of the above multimer. The repeating units of the comb-like composite structure may also be in the form of suitable trihydric alcohols, suitable tetrahydric alcohols, open chain pentitols, open chain hexitols, the corresponding starting materials preferably being in the form of any one of which the hydroxyl groups other than the ether-bonded hydroxyl groups are protected, such as glycerol, trimethylolethane, trimethylolpropane, for example.
Among them, the polyvalent G in the cyclic combination is preferably a residue of a cyclic peptide or a derivative thereof, a residue of a cyclic monosaccharide or a derivative thereof, a residue of a cyclic polysaccharide or a derivative thereof (e.g., a functionalized derivative of cyclodextrin), a skeleton of 1,4, 7-tri-t-butoxycarbonyl-1, 4,7, 10-tetraazacyclododecane, a skeleton of 2-hydroxymethylpiperidine-3, 4, 5-triol, a skeleton of 6-amino-4- (hydroxymethyl) -4-cyclohexyl- [4H,5H ] -1,2, 3-triol, or the like.
For example, when the terminal reactive site k is 4, G is a pentavalent group, including but not limited to the above-mentioned group G 5 A pentavalent group in (1). Pentavalent G can include 1 pentavalent core structure, 1 tetravalent core structure and 1 trivalent core structure, or 3A trivalent nucleus structure. L is 0 -G preferably contains any one of the following structures:
Figure BDA0002963906360000476
Figure BDA0002963906360000481
Figure BDA0002963906360000482
a dendritic structure formed by directly or indirectly combining 3 trivalent G, a comb-shaped structure formed by directly or indirectly combining 3 trivalent G, and the like. Among them, examples of the dendritic structure in which 3 trivalent gs are directly or indirectly combined include a structure in which d is 2 as described above. Comb structure formed by directly combining 3 trivalent groups, including but not limited to a trilysine skeleton, a trimeric glutamic acid skeleton, a trimeric aspartic acid skeleton, a trimeric glycerol skeleton, and the like, such as
Figure BDA0002963906360000483
Figure BDA0002963906360000484
A comb structure formed by indirectly combining 3 trivalent groups, such as three lysines and the like which are combined together by taking amino acids such as glycine, alanine and the like as spacers. Wherein the asterisks within the structure indicate that the asterisk end points to the polyethylene glycol unit.
For example, when the terminal reaction site k ═ 5, G is a hexavalent group, including but not limited to the above set G 6 The hexavalent group of (1). The hexavalent G may include 1 hexavalent core structure, 1 pentavalent core structure and 1 trivalent core structure, 2 tetravalent core structures, 1 tetravalent core structure and 2 trivalent core structures, or 4 trivalent core structures. L is 0 -G preferably contains any one of the following structures: a comb structure composed of 4 trivalent G groups (e.g., tetrapolyglycerol, tetrapolylysine, tetrapolyaspartic acid, tetrapolyglutamic acid, etc.),
Figure BDA0002963906360000485
Figure BDA0002963906360000486
and the like. Wherein the asterisks in the structure indicate that the asterisk ends point to polyethylene glycol units.
2. Preparation method of nitrogen-branched nine-arm polyethylene glycol derivative
2.1. A preparation method of a nitrogen-branched nine-arm polyethylene glycol derivative relates to the following steps:
step one, adopting nine hydroxyl small molecules containing nine hydroxyl groups
Figure BDA0002963906360000491
The initiator system of (1); wherein, deprotonation of nine bare hydroxyl groups forms nonaoxyanions
Figure BDA0002963906360000492
Stably exists under the condition of anionic polymerization;
initiating ethylene oxide polymerization;
step three, adding a proton source into the intermediate product system with nine polyethylene glycol chains obtained in the step two after the reaction is finished, so as to obtain hydroxyl-terminated nitrogen-branched nine-arm polyethylene glycol;
step four, functionalizing the tail end of the nitrogen-branched nine-arm polyethylene glycol to obtain the nitrogen-branched nine-arm polyethylene glycol derivative, wherein the terminal functionalization is selected from any one of terminal linear functionalization and terminal branched functionalization; when g is 0, terminal linear functionalization is performed, and when g is 1, terminal branched functionalization is performed.
The above step may be carried out with or without a solvent, and the solvent is not particularly limited, but is preferably an aprotic solvent such as toluene, benzene, xylene, acetonitrile, ethyl acetate, tetrahydrofuran, chloroform, dichloromethane, dimethyl sulfoxide, dimethyl formamide or dimethyl acetamide, and more preferably dimethyl sulfoxide, dimethylformamide, toluene or tetrahydrofuran.
2.1.1. Nine-pieceHydroxyl small molecule initiator (IN- (OH) 9 )
The structural general formula of the nine-hydroxyl micromolecule initiator is
Figure BDA0002963906360000493
Wherein, N, U 0 、L 2 The definitions of (A) are the same as those in the above general formula (1), and are not described herein again.
The nonahydroxyl small molecule can be a trifunctional small molecule with 1 residue as a nitrogen atom trivalent branching center and 3 residues as U 0 The tetrafunctional micromolecules are obtained through coupling reaction; wherein the residue may be a tri-functional small molecule with a trivalent branching center for the nitrogen atom contains three identical functional groups; the residue may be U 0 The tetrafunctional small molecule contains three or four identical functional groups, when the residue can be U 0 When the tetrafunctional small molecule contains only three identical functional groups, the different functional group ends are connected with the trifunctional small molecule of which the residue can be a nitrogen atom trivalent branching center.
The nine hydroxyl groups of the nine hydroxyl small molecule prepared in the way are almost the same in activity, and the nine hydroxyl groups are respectively connected with U 0 The atom intervals of the branched cores are the same, the nitrogen-branched nine-arm polyethylene glycol derivative further prepared by taking the nine-hydroxyl micromolecule as an initiator molecule has more advantages in purity, the molecular weight and the distribution thereof are more accurately controlled in the product polymerization process, the product has a single structure, the structures of other multi-arm products cannot be generated, the performance is better, the purification difficulty is reduced, the using amount of organic reagents in the purification can be reduced, the cost is reduced, and the nitrogen-branched nine-arm polyethylene glycol derivative is more green and environment-friendly.
The tri-and tetra-functionalized small molecules may be naturally occurring, or purchased directly, or obtained by a suitable coupling reaction. The functional groups participating in the reaction are selected from suitable reactive groups of the class a-class H of the present invention.
In the preparation of the nine-hydroxyl small molecule initiator, the tri-functional small molecule in which the residue can be a nitrogen atom trivalent branching center is preferred
Figure BDA0002963906360000501
Any of the like; the residue may be U 0 The tetrafunctional small molecule of (a) is preferably
Figure BDA0002963906360000502
Figure BDA0002963906360000503
And the like.
The structure of a specific nine-hydroxyl small molecule initiator is exemplified as follows:
Figure BDA0002963906360000504
Figure BDA0002963906360000505
etc.; wherein n is i3 Is 0 or 1, R 1 Is a hydrogen atom or a methyl group.
2.1.2. Initiator system
The structure of the nonahydroxyl small molecule can be characterized and confirmed by the existing conventional technical means, including but not limited to nuclear magnetism, circular dichroism, MALDI-TOF, HPLC, ultraviolet spectrophotometer, FT-IR, mass spectrum, Raman spectrum, single crystal diffraction and the like. The hydroxyl value of the nonahydroxyl micromolecule can be measured by means existing in the related fields, including but not limited to GB/T12008.3-2009 (phthalic anhydride-pyridine method), HG/T2709-95 (acetic anhydride-pyridine method), other acylation methods (such as acetic anhydride-perchloric acid-ethyl acetate catalytic acylation method, acetic anhydride-perchloric acid-dichloroethane catalytic acylation method, acetic anhydride-N-methylimidazole-DMF catalytic acylation method and the like), normal-temperature catalytic acylation method can be adopted, and heating reflux acylation method can also be adopted. GB/T12008.3-2009 or HG/T2709-95 are preferred.
The initiator system also contains alkali, so that the nine-hydroxyl small molecule is deprotonated under the catalysis of the alkali to form oxygen anions.
Deprotonation is carried out under basic conditions. The base used for deprotonation is not particularly limited, but is preferably metallic sodium, potassium, sodium hydride, potassium hydride, sodium methoxide, potassium methoxide, lithium naphthalene, n-butyllithium, t-butyllithium, potassium t-butoxide or diphenylmethylpotassium, more preferably metallic sodium, potassium or diphenylmethylpotassium, and most preferably diphenylmethylpotassium. The catalyst is used in an amount of 5 to 320 mol%, preferably 5 to 80 mol%. If the amount of the catalyst used is less than 5 mol%, the polymerization rate is slow and the cumulative heat increases, resulting in the formation of by-products such as vinyl ether compounds by elimination of the terminal hydroxyl group. In the reaction in the absence of a solvent, the amount of the catalyst exceeding 50 mol% may result in an increase in the viscosity of the reaction solution or precipitation of solids, resulting in imbalance of the reaction and difficulty in purification. When toluene or tetrahydrofuran is used as a solvent, the problem of viscosity increase or solid precipitation of a reaction solution can be solved, and the amount of the catalyst can be correspondingly increased to 80 mol% or more.
Deprotonation is generally carried out at from 10 ℃ to 50 ℃ and preferably from 25 ℃ to 50 ℃. When the temperature is less than 10 ℃, deprotonation is incomplete, and the base participates in anionic polymerization as a nucleophile to obtain low molecular weight impurities with the target polymer chain being 0.5 times of the target molecular weight. Such impurities may react with biologically relevant substances and alter their physical properties. If the species used to initiate the polymerization of ethylene oxide contains protecting groups, temperatures above 50 deg.C can result in partial cleavage and deprotection of the protecting groups to yield high molecular weight impurities above the target molecular weight of the polymer chain of interest. When the drug is modified in a state containing such impurities, the drug preparation is inevitably uneven and unstable in quality, and modification of a high-purity drug cannot be satisfied.
The deprotonation time, preferably from 10 minutes to 24 hours, varies with the base. In general, strong bases with weak basicity or relatively small solubility in organic solvents (such as sodium methoxide, potassium methoxide, sodium hydride, potassium hydride, and the like) require longer deprotonation time, generally 1 to 24 hours; on the other hand, bases having strong basicity and good solubility in organic solvents (e.g., diphenylmethyl potassium, n-butyl lithium, t-butyl lithium, etc.) are sufficiently miscible with the initiator even in the absence of a solvent, and have a high deprotonation rate, generally in the range of 10 minutes to 24 hours, preferably 20 minutes to 1 hour. When deprotonation time is short and deprotonation is incomplete, alkali is used as a nucleophilic reagent to participate in anionic polymerization to obtain low-molecular-weight impurities with target molecular weight of 0.5 time of a target polymer chain; if the material used to initiate the polymerization of ethylene oxide contains a protecting group, then a deprotonation time greater than 24 hours will result in partial cleavage and deprotection of the protecting group to yield a high molecular weight impurity above the target molecular weight; modification of a drug in a state containing such impurities cannot satisfy modification of a high-purity drug.
When potassium methoxide, potassium tert-butoxide, sodium methoxide are used as the catalyst, potassium methoxide is preferred, and the amount thereof is 5 mol% to 80 mol%, and the reaction is carried out at 25 ℃ to 80 ℃, preferably 50 ℃ to 60 ℃, except that, in addition, the reaction is conducted under reduced pressure to promote proton exchange. Because potassium methoxide, potassium tert-butoxide or sodium methoxide itself will also polymerize with ethylene oxide under the polymerization conditions, the end-etherified polyethylene glycol with the target polymer chain molecular weight 0.5 times of the target molecular weight is obtained, interfering the subsequent reaction to generate byproducts. Such reactions require the removal of the lower alcohol by operation at reduced pressure while ensuring complete protonation at higher temperatures (preferably 50 ℃ to 60 ℃).
2.1.3. Polymerization of ethylene oxide
The amount of ethylene oxide used is determined by the design molecular weight of the polyethylene glycol chain, and the metered amount of ethylene oxide is added.
When the polymerization is carried out under aprotic solvent conditions, it is preferably carried out at 50 ℃ to 70 ℃. When the temperature is lower than 50 ℃, the molecular weight is gradually increased along with the polymerization, the viscosity of reaction liquid is increased or solids are separated out, so that the reaction system is not uniform, and the obtained target product is wide in distribution and is not suitable for modification of high-purity medicaments; when the temperature is higher than 70 ℃, the reaction system is easy to explode or generate side reactions, such as the elimination of terminal alcohol to obtain vinyl ether.
When the polymerization is carried out in the absence of a solvent, it is preferably carried out at 50 ℃ to 130 ℃, more preferably at 80 ℃ to 110 ℃. When the temperature is lower than 50 ℃, the polymerization rate is lower, and the accumulated heat is increased, so that the quality of the target product is reduced; in addition, when the temperature is higher than 130 ℃, side reactions such as elimination of a terminal alcohol are liable to occur to give a vinyl ether. Also, as the polymerization proceeds, the molecular weight gradually increases, the viscosity of the reaction liquid may increase or solidification may occur, so that the reaction is not uniform and the distribution of the target product obtained is broad.
The polymerization process may be carried out in a solvent or without a solvent, and the solvent is not particularly limited, but is preferably an aprotic solvent such as toluene, benzene, xylene, acetonitrile, ethyl acetate, tetrahydrofuran, chloroform, dichloromethane, dimethyl sulfoxide, dimethyl formamide or dimethyl acetamide. In general, preference is given to working in aprotic solvents, preferably in dimethyl sulfoxide, dimethylformamide, toluene or tetrahydrofuran.
2.1.4. The polymerization reaction is ended
The polymerization product obtained after the second step is a mixture of alcohol and oxygen anions. When polymerized to a certain extent, a proton source is added to give a hydroxyl-terminated intermediate compound having a specific degree of polymerization. Wherein the proton source is required to provide active hydrogen.
The protonating agent is not particularly limited, and is preferably selected from water, acetic acid, ethanol, and methanol. In general, the amount of protonating agent used is from 1 to 100 times, preferably from 2 to 10 times, that of the deprotonating agent. If the amount of the protonating agent is less than 1 time of the molar equivalent of the deprotonating agent, incomplete protonation can be caused, active oxygen anions can cause unstable product structure, and impurities with molecular weight larger than the target molecular weight can be formed after the product is placed in the air for a long time, so that the molecular weight distribution is widened. When the amount of the protonating agent is more than 20 times the molar equivalent of the deprotonating agent, an excess of the agent or compound causes troubles in purification and may cause side reactions.
The protonation is generally carried out at from 30 ℃ to 100 ℃, preferably from 50 ℃ to 80 ℃. When the temperature is less than 30 ℃, the high molecular weight polymerization product is liable to cause uneven stirring of the reaction system due to viscosity increase or solidification, resulting in incomplete protonation of the product. When the temperature is higher than 100 ℃, the product is easy to chain transfer, so that the molecular weight is increased and the distribution is widened.
The protonation time is preferably from 10 minutes to 60 minutes, the control of which varies depending on the acidity of the protonating agent. Generally, under the condition of weak acidity or two-phase reaction, protonation needs longer time, generally between 30 minutes and 60 minutes; the protonation speed is higher under the condition of stronger acidity or homogeneous reaction, and can be finished within 10 to 30 minutes generally.
2.2. A method for preparing nitrogen branched nine-arm polyethylene glycol derivatives relates to nine functionalized small molecules containing nine same functional groups
Figure BDA0002963906360000521
Reacting with 9 linear double-end functionalized PEG derivative bilPEG molecules to obtain a nitrogen-branched nine-arm polyethylene glycol derivative; wherein bilPEG is monodisperse or polydisperse; wherein functional groups at both ends of bilPEG may be the same or different; wherein, F 1 Contains a reactive functional group capable of reacting with a terminal functional group in the biliPEG to form a divalent linking group L 2 ；
The functional group at the other end of the bilPEG is the same as or different from the target structure; when different, the method further comprises branching the nitrogen into nine-arm polyethylene glycol
Figure BDA0002963906360000522
The terminal functionalization process of (a), the terminal functionalization being selected from any one of terminal linear functionalization and terminal branched functionalization; when g is 0, terminal linear functionalization is carried out, and when g is 1, terminal branched functionalization is carried out.
2.2.1. Nonine functionalized small molecules
The degradability of the nonafunctional small molecule is not particularly limited, and may contain only the stable linker STAG, or 1 or more degradable linkers DEGG. Among them, the kind of the degradable linking group may be 1 or more.
A nine-functional small molecule contains 9 identical reactive groups selected from suitable reactive groups in class a-class H of the present invention. The nine-functional small molecule can be a three-functional small molecule with 1 residue which can be a nitrogen atom trivalent branching center and 3 residues which can be U 0 The tetrafunctional micromolecules are obtained through coupling reaction; wherein the residue may be a tri-functional small molecule with a trivalent branching center for the nitrogen atom contains three identical functional groups; the residue may be U 0 The tetrafunctional small molecule contains three or four identical functional groups, when the residue can be U 0 When the tetrafunctional micromolecule only contains three same functional groups, the different functional group ends are connected with the trifunctional micromolecule of which the residue can be a nitrogen atom trivalent branching center; the nine terminal functional groups of the nine functionalized micromolecules obtained in the way have almost the same activity, so that the nitrogen-branched nine-arm polyethylene glycol derivatives can be endowed with accurate molecular weight and narrower molecular weight distribution in the subsequent coupling reaction with 9 linear polyethylene glycols, and in the subsequent purification process, the difficulty of purification and separation can be reduced, the using amount of organic reagents is reduced, the cost is reduced, and the nine terminal functional groups are more green and environment-friendly.
The residue can be a tri-functional small molecule with a nitrogen atom trivalent branching center and the residue can be U 0 The tetrafunctional small molecule of (a) may be naturally occurring, or purchased directly, or obtained by a suitable coupling reaction. The functional groups participating in the reaction are selected from suitable reactive groups of the class a-class H of the present invention.
Among them, the tri-functional small molecule in which the residue may be a nitrogen atom trivalent branching center is preferable
Figure BDA0002963906360000531
Figure BDA0002963906360000532
Figure BDA0002963906360000533
Any of the like; the residue may be U 0 The tetrafunctional small molecule of (2) is preferably
Figure BDA0002963906360000534
Figure BDA0002963906360000535
Figure BDA0002963906360000536
And the like.
The nine functional small molecule except for the 2.1.1. any nine hydroxyl small molecule initiator IN- (OH) described IN the above 2.1.1 for use IN polymerization 9 In addition, the compound can be selected from any one of the following structures:
Figure BDA0002963906360000541
Figure BDA0002963906360000551
Figure BDA0002963906360000552
etc. wherein n i3 Is 0 or 1, R 1 Is a hydrogen atom or a methyl group.
2.2.2. Linear double-end functionalized PEG derivative bilPEG
The bilPEG has a single functional group at one end capable of reacting with a nonafunctionalized small molecule and is coupled to form a divalent linking group L 2 。
The other end of the bilmpeg, which is the same as or different from the target structure, may have the same or different functional group as the target functional group. Corresponding to general formula (1) can be a PEG terminal hydroxyl group, a linear functionalized functional group (containing only one functional group) or a branched functionalized functional group (which can contain 2 or more functional groups). The functional group carried by the terminal is selected from the group consisting of, but not limited to, any one of the functional groups A to J, and includes a precursor of any one of the reactive groups, a variant thereof as a precursor, a substituted variant, a protected variant, a deprotected variant, and the like. When the terminal contains a reactive group, it preferably contains only one reactive group. For example, when the terminal is terminated with lysine, glutamic acid, aspartic acid, it may be the case that the terminal contains both carboxyl or amino groups, but by subsequent selective protection, it is finally achieved that the terminal contains only one reactive group. Also for example, it is permissible for the end to contain two or more kinds of protected reactive groups, but in subsequent applications, when used to modify a biologically relevant substance, only one of them is selectively deprotected to give a single kind of reactive group. When the structure and the functional group at the end are different from the target, the end of the nitrogen-branched nine-arm polyethylene glycol derivative obtained by the coupling reaction can be functionalized to obtain the target structure with the target functional group. Suitable terminal linear functionalization or terminal branched functionalization can be carried out. When the terminal is a linear structure or a branched structure as the target structure and differs only in the functional group at the terminal, the target functional group can be obtained preferably by deprotection alone.
The functional groups at both ends of the bilipeg may be the same or different, and a linear heterofunctional PEG derivative (bisheterolpeg) having two different functional groups at both ends is preferable. Pairs of heterofunctional groups that may be present simultaneously include, but are not limited to: hydroxyl and protected hydroxyl, hydroxyl or protected hydroxyl and non-hydroxyl reactive groups of class A-class H (such as amino, protected amino, amine salt, aldehyde group, active ester group, maleimide group, carboxyl, protected carboxyl, alkynyl, protected alkynyl, azide group, alkenyl, acrylate group, methacrylate group, epoxy group, isocyanate group and the like), hydroxyl or protected hydroxyl and functional groups or derivatives thereof (such as targeting group, photosensitive group and the like) in class I-class J, active ester group and maleimide group, active ester group and aldehyde group, active ester group and azide group, active ester group and alkyne group or protected alkyne group, active ester group and acrylate group, active ester group and methacrylate group, active ester group and acrylate group, maleimide group and azide group, hydroxyl or protected hydroxyl group, Maleimide and alkyne or protected alkyne, maleimide and acrylate, maleimide and methacrylate, maleimide and acrylate, maleimide and carboxyl, maleimide and amino or protected amino or amine, maleimide and isocyanate, maleimide and protected thiol, aldehyde and azide Base, aldehyde group and acrylate group, aldehyde group and methacrylate group, aldehyde group and acrylate group, aldehyde group and epoxy group, aldehyde group and carboxyl group, aldehyde group and alkynyl group or protected alkynyl group, azido group and sulfhydryl group or protected sulfhydryl group, azido group and amino group or protected amino group or amine salt, azido group and acrylate group, azido group and methacrylate group, azido group and acrylate group, azido group and carboxyl group, acrylate group and amino group or protected amino group or amine salt, acrylate group and isocyanate group, acrylate group and epoxy group, acrylate group and methacrylate group, acrylate group and carboxyl group, methacrylate group and amino group or protected amino group or amine salt, methacrylate group and isocyanate group, methacrylate group and epoxy group, alkynyl group or protected alkynyl group and amino group or protected amino group or amine salt, Alkynyl or protected alkynyl and isocyanate group, alkynyl or protected alkynyl and acrylate group, alkynyl or protected alkynyl and methacrylate group, alkynyl or protected alkynyl and acrylate group, alkynyl or protected alkynyl and epoxy group, alkynyl or protected alkynyl and carboxyl group, protected alkynyl and azido group, acrylic group and isocyanate group, acrylic group and acrylate group, acrylic group and epoxy group, acrylic group and carboxyl group, carboxyl group and mercapto group or protected mercapto group, carboxyl group and amino group or protected amino group or amine salt, carboxyl group and isocyanate group, carboxyl group and epoxy group, amino group or protected amino group or amine salt and mercapto group or protected mercapto group, targeting group and non-hydroxyl reactive group, photosensitive group and non-hydroxyl reactive group, etc. Wherein the active ester includes, but is not limited to, any one of the active esters of succinimide (such as succinimide carbonate), p-nitrophenyl active ester, o-nitrophenyl active ester, benzotriazole active ester, 1,3, 5-trichlorobenzene active ester, 1,3, 5-trifluorobenzene active ester, pentafluorobenzene active ester, imidazole active ester, 2-thiothioxothiazolidine-3-carboxylate, 2-thiopyrrolidine-1-carboxylate, etc.; the amino group includes primary and secondary amino groups. The amine salt is preferably in the form of the hydrochloride salt of an amino group such as NH 2 HCl。
The bilmpeg may be polydisperse or monodisperse. When polydisperse, corresponding to n 1 ≈n 2 ≈n 3 ≈n 4 ≈n 5 ≈n 6 ≈n 7 ≈n 8 ≈n 9 When monodisperse, corresponds to n 1 ＝n 2 ＝n 3 ＝n 4 ＝n 5 ＝n 6 ＝n 7 ＝n 8 ＝n 9 。
When the bilmpeg is polydisperse, its polydispersity is not particularly limited, but a raw material having a polydispersity PDI of less than 1.15, more preferably less than 1.10, more preferably less than 1.08, more preferably less than 1.05 is preferable. The lower the PDI, the more uniform the molecular weight, and the narrower the molecular weight distribution of the resulting product, and the higher the quality of the modified product when used to modify molecules such as drugs, the more practical the demand.
When the bilmpeg is monodisperse, PDI is 1, nine PEG chains each have a fixed molecular weight and are equal to each other, and a nitrogen-branched nine-arm polyethylene glycol derivative having a definite molecular structure can be obtained. When the product is used for modifying the medicine, the product with a determined medicine structure can be obtained, and the performance is more uniform and easy to control.
The product prepared by adopting the monodisperse raw material has uniform molecular weight, but the molecular weight is more limited and the steps are long due to the limitation of the preparation method. The advantages of using polydisperse starting materials are simple procedure and large molecular weight adjustment range.
The preparation method for preparing monodisperse polyethylene glycol chains can adopt the known techniques in the technical field, including but not limited to the documents J, org, chem.2006,71, 9884-.
2.3. End functionalization
The process of modifying a hydroxyl group or a non-target functional group located at the end of a polyethylene glycol chain into a target functional group is end functionalization, which can be classified into end linear functionalization and end branched functionalization. The functional group includes, but is not limited to, the functional groups listed in class A to class J. The polyethylene glycol after the initiation of ethylene oxide polymerization has a hydroxyl group at the end, and a hydroxyethyl group is formed at the end corresponding to g ═ 0 and k ═ 1.
In the general formula (1), the end-functionalization process when G in F is 0 is end-linear functionalization, in which case the corresponding G is absent, k is 1, and the functional group R at the end of the polyethylene glycol chain is 01 The number of (2) is 1; the end functionalization process when G in F is 1 is end branching functionalization, k is 2-250, the corresponding G is a branched group with a valence of k +1, and a functional group R at the tail end of the polyethylene glycol chain 01 The number of (2) is k.
When k in F is 1, performing terminal linear functionalization; and when k in F is greater than 1, performing terminal branching functionalization.
2.3.1. Linear functionalization of polyethylene glycol chain ends
The method of terminal linear functionalization is not particularly limited, depending on the type of the final functional group. The functionalization may be linear functionalization based on the terminal hydroxyl group of the polyethylene glycol chain, or conversion to a target functional group based on a reactive group, or a combination of both. Techniques known in the art may be used. And include, but are not limited to, those described and exemplified in documents CN104530413A, CN104530415A, CN104530417A, CN 104877127A. For example, CN104530413A corresponds to segments [960] to [1205], and for example CN104877127A corresponds to segments [1005] to [1087 ]. The reaction temperature, reaction time, dosage, solvent condition, reaction condition (such as strong basicity and acidity), catalyst, deprotonation reagent, oxidant, reducing agent, alkylation reagent, halogenated reagent, weak acidic salt and other parameters and the optimization of each parameter are well known to those skilled in the art, or can be obtained by optimization through limited experiments, and are not repeated herein, but the reaction principle, reaction raw materials, reaction routes and the like of the involved reaction types (such as condensation reaction, ring-opening reaction, ring-closing condensation reaction, esterification reaction, oxidation reaction, addition reaction, substitution reaction, alkylation reaction, dehydrogenation reaction and the like) are mainly described in general, and the relevant details or the preferred conditions are also well known to those skilled in the art and can be obtained through limited experiments.
2.3.1.1. Class A: r 01 Functionalization selected from class A
The functional group in class A is an active ester or an analog of an active ester. The preparation method includes, but is not limited to, the following methods.
a: the corresponding active esters (A6-A10, A12, A14) can be obtained by condensation reaction of a terminal hydroxyl intermediate with corresponding carbonates (N, N '-disuccinimidyl carbonate, di (p-nitrophenyl) carbonate, di (o-nitrophenyl) carbonate, bitertazole carbonate, etc.), haloformates (p-nitrophenyl chloroformate, o-nitrophenylchloroformate, trichlorophenylchloroformate, etc.), N' -carbonyldiimidazole in the presence of a base. The corresponding ring-substituted derivatives of hydrogen atoms can also be obtained in a similar manner, e.g. by reaction with 1, 1' -carbonylbis (2-methylimidazole) to give the active ester of 2-methylimidazole. The corresponding haloformate is selected from chloro, bromo or iodo, preferably chloro.
b: the corresponding active esters (A1-A5, A11, A13) can also be obtained by condensation reactions. The terminal hydroxyl group is reacted in one step or multiple steps to obtain terminal carboxyl group, and then the terminal carboxyl group is reacted with corresponding alcohol (N-hydroxysuccinimide, p-nitrophenol, o-nitrophenol, trichlorophenol, 1-hydroxybenzotriazole and the like) in the presence of a condensing agent to obtain corresponding active ester.
c: analogs of the active esters (A15-A18) can be prepared by reacting the terminal carboxyl group with the corresponding amine (e.g., thiazole-2-thione, pyrrolidine-2-thione, benzo [ d ] thiazole-2 (3H) -thione, 4-oxo-2-thioxothiazolidine, etc.) in the presence of a condensing agent to give the corresponding amide. The corresponding derivatives in which the hydrogen atom of the ring is substituted can also be obtained in an analogous manner, for example by reacting with 4-isopropyl-1, 3-thiazolidine-2-thione, (R) -4-isopropylthiazoline-2-thione, 4-phenylthiazoline-2-thione, etc., to give the corresponding active ester analogs.
The condensing agent is not particularly limited, but N, N ' -Dicyclohexylcarbodiimide (DCC), 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC. HCl), 2- (7-azobenzotriazol) -N, N ' -tetramethyluronium Hexafluorophosphate (HATU), benzotriazol-N, N ' -tetramethyluronium Hexafluorophosphate (HBTU), and most preferably DCC, are preferable. The solvent may be a non-solvent or an aprotic solvent. The base includes generally organic bases, preferably triethylamine, pyridine.
2.3.1.2. Class B: r 01 Functionalization selected from class B
The sulfonic acid or sulfinic acid ester derivatives (B1, B2) can be separated from the leaving group Y through a terminal hydroxyl group 1 The sulfonyl chloride and the sulfinyl chloride are esterified in the presence of alkali to obtain the product. Y is 1 The definition of (c) is consistent with the above. The solvent may be a non-solvent or an aprotic solvent. The base includes organic or inorganic bases, preferably organic bases, more preferably triethylamine, pyridine.
The sulfone or sulfoxide derivative (B3, B4) can be modified by containing a leaving group Y 1 The sulfoxide intermediate or thioether intermediate is prepared by oxidation reaction. Y is 1 The definitions of (a) and (b) are consistent with the above. The oxidizing agent is not particularly limited as long as it is a compound or a combination of compounds capable of raising the valence of the substrate. The solvent may be a non-solvent or an aprotic solvent.
The sulfone derivative (B3) can be obtained by deprotonation of the terminal hydroxyl group by reaction with a base, and addition reaction with vinyl sulfone.
The disulfonyl derivative (B5) and its modified forms (B6) can be prepared by the methods of the reference { Advanced Drug Delivery Reviews, 2008,60,3-12 }.
2.3.1.3. Class C: r 01 Functionalization selected from class C
The hydroxylamine compound (C1) can be obtained by reacting a terminal hydroxyl group with an excess of hydroxylamine hydrochloride under strongly basic conditions (e.g., diphenylmethyl potassium).
The mercapto derivative (C2) can be obtained by reacting a terminal hydroxyl group with thiourea, and the reaction can be carried out in a solvent or without a solvent, and the solvent is not limited, and is preferably water, toluene, benzene, xylene, acetonitrile, ethyl acetate, diethyl ether, methyl t-butyl ether, tetrahydrofuran, chloroform, dichloromethane, dimethyl sulfoxide, dimethylformamide or dimethylacetamide, preferably water, tetrahydrofuran, dichloromethane, acetonitrile.
The mercapto derivative (C2) can also be obtained by reacting a sulfonate compound with a potassium xanthate compound, followed by decomposition with a primary amine. This reaction can be carried out without solvent or with a solvent, which is not limited, and an aprotic solvent is preferred.
The protected sulfur compound (C7) can be prepared by reacting the corresponding sulfur compound (C2) with a corresponding protecting agent. The method of preparation is not limited, including but not limited to the following methods: a. thioether-protected sulfides can be prepared by reacting a sulfur compound with a corresponding alkylating agent having a leaving group in the presence of a base. The solvent may be a non-solvent or an aprotic solvent. b. Thioesters (C7& C17) can be prepared from sulfur compounds by reaction with the corresponding acid halides in the presence of a base. The solvent may be a non-solvent or an aprotic solvent.
The amine derivative (C4) can be subjected to coupling reaction of terminal hydroxyl and acrylonitrile or the like under the catalysis of alkali, and then the cyano can be reduced in an autoclave under the catalysis of palladium or nickel to obtain the corresponding amine. This reaction can be carried out without solvent or under solvent conditions, the solvent being not limited, preferably water or 1, 4-dioxane and combinations thereof. The base includes organic base or inorganic base, preferably inorganic base, more preferably sodium hydroxide, potassium hydroxide.
The amine derivative (C4) can also be obtained by reacting a sulfonate compound (B1) with aqueous ammonia.
Protected amine compounds (C6) can be prepared by reacting the corresponding amine (C3) with the corresponding protecting reagent. The method of preparation is not limited, and includes, but is not limited to, the following methods:
a. the carbamate compounds can be prepared by reacting amines with the corresponding haloformates in the presence of a base. The solvent may be a non-solvent or an aprotic solvent. The base includes organic or inorganic bases, preferably organic bases, more preferably triethylamine, pyridine.
b. The amide compound can be prepared by reacting amine with corresponding acyl halide in the presence of alkali.
c. Alkylated amino compounds can be prepared by reacting an amine with a corresponding alkylating agent having a leaving group in the presence of a base. The solvent may be an aprotic or non-protic solvent base including organic or inorganic bases, preferably organic bases, more preferably triethylamine, pyridine, sodium hydride, DPMK, potassium hydride, sodium alkoxide.
d. Another process for the preparation of alkylated amine compounds is that of reacting an amine with the corresponding aldehyde or ketone to give an imine compound and then reducing the imine (Schiff base) in the presence of a reducing agent to give the corresponding alkylated amine compound (C5). The corresponding aldehyde or ketone is not particularly limited. The solvent may be a protic solvent or an aprotic solvent, and the solvent includes toluene, benzene, xylene, acetonitrile, ethyl acetate, diethyl ether, methyl tert-butyl ether, tetrahydrofuran, methanol, ethyl acetate, dimethylformamide or dimethylacetamide, preferably tetrahydrofuran, methanol, ethyl acetate. The reducing agent is not particularly limited as long as it can reduce the Schiff base formed from ammonia and aldehyde or ketone to an amino group; preferably one or a combination of sodium borohydride, sodium cyanoborohydride, lithium aluminum hydride, borane, diborane, diisobutylaluminum hydride, diisopinocampheylborane, lithium borohydride, zinc borohydride, borane-pyridine, borane-methyl sulfide, borane-tetrahydrofuran, and the like; more preferably sodium cyanoborohydride.
The halide (C7), the tetramethylpiperidinyloxy compound (C8) and the dioxopiperidinyloxy compound (C9) can be obtained by reacting the sulfonate compound (B1) with the corresponding halide salt, 2,6, 6-tetramethylpiperidine-nitrogen-hydroxy group and 3, 5-dioxo-1-cyclohexylamine. The bromine salt is not limited as long as a free bromide ion is generated in the solvent, and sodium bromide and potassium bromide are preferable.
The halogenated compound (C7) can also be obtained by reacting a terminal hydroxyl group with a halogenating agent. The halogenating agent is not particularly limited as long as it can convert a hydroxyl group into a corresponding halogen atom, and is preferably one or a combination of thionyl chloride, phosphorus trichloride, phosphorus tribromide, dibromosulfoxide, and the like. The solvent may be a non-solvent or an aprotic solvent.
The ester and thioester compounds (C17) can be obtained by condensation reaction of a terminal hydroxyl group, a mercapto group and a terminal carboxyl group or an acid halide, preferably an acid chloride.
Thioester compounds (C7) can also be obtained by reaction between a sulfhydryl group and an activated ester. Reference is made to the document { Journal of Controlled Release,2014,194: 301-.
The carbonate or thiocarbonate compound (C18) can be obtained by condensation reaction of a terminal hydroxyl group, a mercapto group and an oxycarbonylchloride compound. Such as ethyl chloroformate, ethyl thiocarbonate.
The trithiocarbonate derivative (C18) can also be obtained by coupling reaction of a trithiocarbonate-containing small molecule compound (such as 3- (benzylthiocarbonylthio) propionic acid) with a functionalized polyethylene glycol carrying a suitable functional group.
The ester compound (D11) is treated with ammonia water and hydrazine hydrate to obtain the amide compound (C20) and the hydrazide compound (C21), respectively.
The halogenated acetamide compound (C10) can be obtained by reacting halogenated acetic acid with polyethylene glycol amine derivative (C4) under the action of a condensing agent to generate amide bond.
The lipoic acid derivative (C14) can be obtained by condensation reaction of lipoic acid and corresponding alcohol (H1) or amine (C4).
2.3.1.4. Class D: r 01 Functionalization selected from class D
The ester compounds (D11) and thioester compounds (D26, D27, D28) can be obtained by deprotonating the terminal hydroxyl group and then subjecting the resulting product to substitution reaction with an α -haloester. Examples thereof include ethyl chloroacetate and ethyl bromoacetate.
Thioesters (D26) can also be obtained by reacting the corresponding esters (D11) with thiols.
The ester compound (D11) is hydrolyzed with an alkaline solution to give a carboxylic acid compound (D1).
The acid halide compound (D4) can be obtained by reacting a carboxylic acid compound (D1) with a halogenating agent. The halogenating agent is not particularly limited as long as it can convert a hydroxyl group in the carboxylic acid into a corresponding halogen atom, and is preferably one or a combination of thionyl chloride, phosphorus trichloride, phosphorus tribromide, dibromosulfoxide, and the like. The solvent may be a non-solvent or an aprotic solvent.
The acid anhydride derivative (D11) can be obtained by reacting the carboxylic acid derivative (D4) with an acid halide, a small-molecule acid anhydride, or a small-molecule mixed acid anhydride. The acid halide, the small molecule acid anhydride, and the small molecule mixed acid anhydride reagent are not particularly limited as long as they can convert a carboxylic acid into the corresponding acid anhydride, and C is preferred 1-10 Acid chloride, C 1-10 Acyl bromide, C 1-10 Anhydride, and the like, or combinations thereof.
The sulfonic acid derivative (D2) can be obtained by alkylating a haloalkylsulfonic acid (e.g., 2-bromoethylsulfonic acid) with a terminal hydroxyl group.
The acetaldehyde derivative (D6) can be obtained by direct oxidation of the terminal hydroxyl group. The oxidizing agent is not particularly limited, but preferably pdc (pyridinium chloride), pcc (pyridinium dichloride), DCC + DMSO, oxalyl chloride + MDSO, sulfur trioxide pyridine + DMSO, trifluoroacetic anhydride + DMSO, MnO are used 2 Preferably DCC + DMSO. The reaction solvent is not particularly limited, and an aprotic solvent is preferred. In addition, a weakly acidic salt should be added in this reaction, and there is no particular limitation, but pyridine trifluoroacetate, triethylamine trifluoroacetate, pyridine hydrochloride, triethylamine hydrochloride, pyridine sulfate, triethylamine sulfate and the like are preferable, and pyridine trifluoroacetate is more preferable.
Propionaldehyde and other aldehyde derivatives (D6) can be deprotonated by the terminal hydroxyl group and then reacted with a halide to give an acetal intermediate (D7), and the compound (D7) is hydrolyzed under acidic conditions to give the corresponding aldehyde. The base used for deprotonation is not particularly limited, but is preferably metallic sodium, potassium, sodium hydride, potassium hydride, sodium methoxide, potassium tert-butoxide or diphenylmethyl potassium, and more preferably sodium hydride or diphenylmethyl potassium. The reaction solvent is not particularly limited, and an aprotic solvent is preferred. The acetal deprotection is carried out under acidic conditions, the solution pH preferably being 1 to 4. The acid is not particularly limited, but acetic acid, phosphoric acid, sulfuric acid, hydrochloric acid, nitric acid are preferable, and hydrochloric acid is more preferable. The reaction solvent is not particularly limited as long as it can dissolve the reactants and the product, and water is preferred.
The aldehyde derivative (D6) can also be introduced through a coupling reaction of a small molecule reagent containing an acetal structure and acetal deprotection. For example, the corresponding-C (═ O) - (CH) can be obtained by amidation reaction of polyethylene glycol amine with 2, 2-diethoxyacetic acid, 3-diethoxypropionic acid, 4-diethoxybutyric acid, 5-diethoxypentanoic acid, etc., followed by removal of acetal protection 2 ) 0～ 3 A CHO aldehyde derivative.
The acetal derivative (D7) can also be prepared by reacting polyethylene glycol aldehyde derivative (D6) with corresponding alcohol under the catalysis of acid to obtain polyethylene glycol (D7) in aldehyde protection form. The acid is not particularly limited, and may be a protonic acid or a Lewis acid, and among them, hydrochloric acid, sulfuric acid, trifluoroacetic acid, trifluoromethanesulfonic acid, p-toluenesulfonic acid, aluminum trichloride, stannic chloride and the like are preferable. Among them, protonic acids are preferable, and hydrochloric acid, sulfuric acid, trifluoroacetic acid, trifluoromethanesulfonic acid, phosphoric acid, and nitric acid are more preferable. The protonic acid is preferred, and the alcohol is more preferred, but not particularly limited, and may be a monohydric alcohol, a dihydric alcohol or a polyhydric alcohol, among which methanol, ethanol, propanol, butanol, pentanol, ethylene glycol, 1, 3-propanediol, 1, 4-butanediol, and the like are preferred. The solvent may be a non-solvent or an aprotic solvent.
The isocyanate (D9) and thioisocyanate (D10) derivatives can be obtained by reacting an alcohol (H1) and an amine derivative (C4) with an excess of diisocyanate or dithioisocyanate. The diisocyanate and dithioisocyanate are not particularly limited, and preferably contain C 1-10 Diisocyanate, C 1-10 A dithioisocyanate. The solvent may be a non-solvent or an aprotic solvent. The diisocyanate includes, but is not limited to, 1, 6-hexamethylene diisocyanate, dimethylbiphenyl diisocyanate, methylene-bis-p-phenyl diisocyanate, p-phenylene diisocyanate, toluene-2, 4-diisocyanate, 1, 5-naphthalene diisocyanate, m-xylylene isocyanate, isophorone diisocyanate, 4-diisocyanate dicyclohexylmethane, bis (2-isocyanate) -5-norbornene-2, 3-dicarboxylate.
The oxycarbonylchloride derivative (D12) can be obtained by reacting a terminal hydroxyl group (H1) with triphosgene under an alkaline condition. The base is preferably an organic base, such as dimethylaminopyridine, for example. The solvent is preferably an aprotic solvent, such as, for example, dichloromethane.
Squaric acid ester (D24) can be obtained by the reaction between an amine derivative (C4) and a squaric acid diester.
2.3.1.5. Class E: r 01 Functionalization selected from class E
The maleimide derivative (E1) can be prepared by the ring-opening reaction of an amine compound (C4) and maleic anhydride to obtain an acid intermediate (E5), and then the ring-closing condensation reaction is carried out under the catalysis of acetic anhydride or sodium acetate. The reaction solvent is not particularly limited, and an aprotic solvent is preferred. In the ring-closing condensation reaction, the solvent is not limited, and the above-mentioned aprotic solvent or acetic anhydride is preferable.
The maleimide derivative (E1) can also be obtained by condensation reaction of an amine compound (C3) with an acid or an active ester containing a maleimide group (MAL group). Acids containing MAL groups include, but are not limited to, 3-maleimidopropanoic acid, 4-maleimidobenzoic acid, 6-maleimidocaproic acid, 11-maleimidoundecanoic acid. Active esters containing MAL groups include, but are not limited to, maleimidoacetate succinimidyl ester, 3-maleimidopropionate hydroxysuccinimide ester, 6- (maleimido) hexanoate succinimidyl ester, 3-maleimidobenzoate succinimidyl ester, 4- (N-maleimidomethyl) cyclohexane-1-carboxylate succinimidyl ester, 4- (4-maleimidophenyl) butanoate succinimidyl ester, 11- (maleimido) undecanoate succinimidyl ester, N- (4-maleimidobutyryl) succinimide. Similarly, the diaza-maleimide derivative (E6) can also be obtained by condensation reaction of an amine compound (C3) with a corresponding acid or active ester.
The maleimide derivative (E1) can also be obtained by condensation reaction of an active ester derivative (A1-A14) and an amine containing an MAL group. Amines containing MAL groups include, but are not limited to, N- (2-aminoethyl) maleimide, N- (4-aminophenyl) maleimide.
The α, β -unsaturated esters (E2, E3) can be obtained by deprotonating the terminal hydroxyl group and reacting with the corresponding halo. The deprotonated base is not limited, and is preferably metallic sodium, potassium, sodium hydride, potassium hydride, sodium methoxide, potassium tert-butoxide or diphenylmethyl potassium, and sodium hydride or diphenylmethyl potassium is more preferably used. The reaction solvent is not limited, and an aprotic solvent is preferred. Examples of the halide include acryloyl chloride and methacryloyl chloride.
The derivative (E5) can also be prepared by reacting an amine derivative (C5) with a corresponding dicarboxylic acid in the presence of a condensing agent to obtain a corresponding amide derivative. The condensing agent is not particularly limited, but DCC, EDC · HCl, HATU, HBTU are preferable, and DCC is most preferable. The solvent may be a non-solvent or an aprotic solvent. The base includes generally organic bases, preferably triethylamine, pyridine.
Azo compounds (E7), cyclic compounds containing unsaturated double bonds (E8), norbornene compounds (E9), 2, 5-norbornadiene compounds (E10) and 7-oxa-bicyclo [2.2.1] hept-5-ene compounds (E11) can be obtained by condensation reaction of alcohol, carboxylic acid, amine, amide and methyl ester derivatives containing corresponding ring structures with corresponding reactive groups to generate linking groups including but not limited to ester bonds, amide bonds, urethane bonds, carbonate bonds, hydrazide bonds and the like. Such as cyclooctene-4-methanol of the type (E8), methyl-4-cycloocten-1-yl carbonate, cyclooctene-4-carboxylic acid. Examples of classes such as (E9) include, but are not limited to, 5-norbornene-2-methanol, 5-norbornene-2-carboxylic acid (2-hydroxyethyl) ester, a-dimethylbicyclo [2.2.1] hept-5-ene-2-methanol, 5-norbornene-2-methanamine, 5-norbornene-2-carboxylic acid, 2-methylbicyclo [2.2.1] hept-5-ene-2-carboxylic acid, hydrogen succinate-1- (1-bicyclo [2.2.1] hept-5-ene-2-ethyl) ester, 5-norbornene-2-carboxamide, 2-methylbicyclo [2.2.1] hept-5-ene-2-carboxamide, 2-cyano-5-norbornene, 5-norbornen-2-yl (ethyl) chlorodimethylsilane, N- [4- (4-aminophenyl) phenyl ] -5-norbornene-2, 3-dicarboximide, N-hydroxy-5-norbornene-2, 3-dicarboximide, 5-norbornene-2-carbaldehyde, nadic anhydride, methylendomethylenetetrahydrophthalic anhydride, N-methyl-N-propargyl- (5-norbornadiene-2-methylamine).
2.3.1.6. Class F: r is 01 Selected from class FFunctionalization
The compounds (F1, F2, F3 and F4) can be obtained by deprotonating a terminal hydroxyl and then carrying out substitution reaction with a corresponding halide. The base to be deprotonated is not limited, and is preferably metallic sodium, potassium, sodium hydride, potassium hydride, sodium methoxide, potassium tert-butoxide or diphenylmethyl potassium, more preferably sodium hydride or diphenylmethyl potassium. The reaction solvent is not particularly limited, and an aprotic solvent is preferred. Examples of the halogenated compound corresponding to the epoxy compound (F1) include epichlorohydrin, 2-chloromethyl-2-methyloxirane, 3-chlorophenyloxirane, epifluoropropane, epibromohydrin, 4-bromo-1, 2-epoxybutane, 6-bromo-1, 2-epoxyhexane, etc.; preference is given to epichlorohydrin. Examples of the halogenated compound corresponding to the vinyl compound (F1) include 3-vinyl chloride and 3-vinyl bromide. Examples of the halide corresponding to the acetylene compound include 3-bromopropyne. Examples of the halide corresponding to the protected acetylene compound include 3-methylsilylbromopropyne and 3-t-butyldimethylsilylbromopropyyne.
2.3.1.7. Class G: r 01 Functionalization selected from class G
Cycloalkyne compounds (G1-G7), cyclodiolefine compounds (G8, G9) and furan compounds (G10) can be obtained by condensation reaction of alcohol, carboxylic acid, amine, amide and methyl ester derivatives containing corresponding ring structures with corresponding reactive groups to generate linking groups including but not limited to ester bonds, amide bonds, carbamate bonds, carbonate bonds, hydrazide bonds and the like. The following are exemplified as raw materials:
Figure BDA0002963906360000611
Figure BDA0002963906360000612
And the like.
The azide compound (G21) can be obtained by reacting the sulfonate compound (B1) with the corresponding azide salt. The azide salt is not limited as long as a free azide ion is generated in the solvent, and sodium azide and potassium azide are preferable. The solvent for the reaction is not limited, and is preferably carried out in a solvent of water, ethanol, acetonitrile, dimethyl sulfoxide, dimethylformamide or dimethylacetamide, preferably water and dimethylformamide.
The cyanohydrin (G22) can be obtained by oxidizing an aldehyde derivative (D6) and hydroxylamine to form an oxime (G24). The oxime is formed in a solvent which may be a non-solvent or an aprotic solvent. In the oxidation process, the oxidizing agent is not particularly limited, and is preferably one or a combination of N-iodosuccinimide, N-chlorosuccinimide, N-bromosuccinimide, and the like. The solvent may be a non-solvent or an aprotic solvent.
The nitrile compound (G23) can be obtained by an addition reaction between a terminal hydroxyl group and acrylonitrile under basic conditions. Or the derivative can be obtained by pressurizing the amine derivative (C4) with ammonia and then hydrogen under the catalysis of nickel or palladium carbon, and then carrying out dehydrogenation reaction under the high-temperature condition.
Compounds (G31) and (G32) can be prepared using the methods of the literature PCT/US2013/046,989.
2.3.1.8. Class H: r 01 Functionalization selected from class H
The product obtained after initiating the polymerization of ethylene oxide is a mixture of alcohol and oxyanion, and is protonated to obtain a polyethylene glycol chain with terminal hydroxyl groups (H1).
The hydroxyl-terminated alcohol derivative (H1) may also be obtained by modifying a non-hydroxyl-reactive group, for example, by reacting ethylene carbonate with a secondary amine to form-NH-CH (═ O) CH 2 CH 2 An alcohol of OH structure.
The hydroxyl-terminated alcohol derivative (H1) can also be obtained by diazotizing an amine derivative (C4) with a nitrite and hydrolyzing under low-temperature acidic conditions. The acid is not particularly limited, and may be a protonic acid or a Lewis acid, among which protonic acids are preferable, and hydrochloric acid, sulfuric acid, trifluoroacetic acid, trifluoromethanesulfonic acid, phosphoric acid, and nitric acid are more preferable. The low temperature is preferably about 0 ℃.
The protected hydroxyl group (H2) can be obtained by reacting a terminal hydroxyl group with a protecting agent, and the protecting agent is not particularly limited, but a halogenated silane, a carboxylic acid, an acid chloride, an acid anhydride, a halogenated hydrocarbon, a sulfonyl chloride, an alkenyl ether, a carbonyl group, and the like are preferable.
A. Typically, the terminal hydroxyl group is inReacting with halosilanes, acid chlorides, acid anhydrides, sulfonyl chlorides, halogenated hydrocarbons under acidic conditions or in the presence of a base to give the protected form (H2). The solvent may be a non-solvent or an aprotic solvent. The base includes an organic base or an inorganic base, preferably an organic base, more preferably triethylamine, pyridine. Protected forms of the ether structure OPG 4 In accordance with the foregoing.
B. The terminal hydroxyl group is reacted with a carboxylic acid in the presence of a base and a condensing agent under reaction conditions to produce (H2) 01 The process for preparing active esters selected from class A is similar.
C. The terminal hydroxyl group is subjected to addition reaction with an alkenyl ether in the presence of an acid to give (H2), and the alkenyl ether is not particularly limited, but ethyl vinyl ether and tetrahydropyran are preferable. The acid is not particularly limited, and may be a protonic acid or a Lewis acid. The solvent may be a non-solvent or an aprotic solvent.
D. The terminal hydroxyl group and tert-butyldimethylchlorosilane, vinyl ether, dihydropyran, benzyl bromide and di-tert-butyl dicarbonate can be respectively protected by silicon group, vinyl ether group, dihydropyran group, benzyl group and Boc.
The terminally protected dihydroxyl group (H3) can be obtained by a method including, but not limited to, the references { Macromol. biosci.2011,11, 1570-1578}, the references { J.Am. chem. Soc., Vol.123, No.25,2001 }.
The photoreactive groups (H6) and (H7) which can be converted into enolic hydroxyl groups can be prepared by the method of document U.S. Pat. No. 3, 14,021,040.
2.3.1.9. Class I: r 01 Functionalization selected from class I
The pegylated folic acid (I1) can be obtained by condensation reaction of carboxyl group in folic acid with polyethylene glycol alcohol or its alcohol derivative (H1) and amine derivative (C4), and among them, the condensation agent is not particularly limited, but DCC, EDC HCl, HATU, HBTU are preferable, and DCC is most preferable. While the condensing agent is generally 1 to 20 times, preferably 5 to 10 times, the molar equivalent of folic acid, and a suitable catalyst (e.g., 4-dimethylaminopyridine) may be added for this reaction. The solvent may be a non-solvent or an aprotic solvent. The base includes generally organic bases, preferably triethylamine, pyridine.
The pegylated cholesterol (I2) can be obtained by condensation reaction of the terminal hydroxyl group of polyethylene glycol with carboxylic acid (D1), acid halide (D4), sulfonyl chloride (D5), isocyanate (D9), isothiocyanate (D10), and the like. Pegylated cholesterol can also be obtained by coupling reaction of cholesterol derivatives with appropriate groups. Taking cholesteryl hydrogen succinate as an example, the compound can be obtained by condensation reaction with the terminal hydroxyl of polyethylene glycol.
The pegylated biotin (I3) can be obtained by condensation reaction of carboxyl group in biotin with polyethylene glycol or its alcohol derivative (H1) and amine derivative (C4). The reaction conditions are the same as those described above for the reaction between carboxyl groups and hydroxyl groups. Biotin derivatives such as D-desthiobiotin and 2-iminobiotin can also be obtained by condensation of carboxyl groups with polyethylene glycol or its alcohol derivatives (H1) and amine derivatives (C4).
The pegylated biotin (I3) can also be obtained by coupling reaction of biotin derivatives including but not limited to those disclosed above in the present invention with a suitable polyethylene glycol or its derivative selected from the group consisting of polyethylene glycol, amine derivatives of polyethylene glycol (C4), alkyne derivatives (F3, G1-G7), carboxylic acid derivatives (D1), acid halide derivatives (D4), aldehyde derivatives (D6), and the like. Wherein, the amine derivative and the alcohol derivative of the biotin can also be subjected to alkylation reaction with corresponding polyethylene glycol sulfonate or polyethylene glycol halide.
2.3.1.10. Class J: r 01 Functionalization selected from class J
Fluorescein and its derivatives (including but not limited to J1, J3), rhodamine and its derivatives (including but not limited to J2), anthracene and its derivatives (J4), pyrene and its derivatives (J5), coumarin and its derivatives (including but not limited to J6), fluorescein 3G and its derivatives (including but not limited to J7), carbazole and its derivatives (J8), imidazole and its derivatives (J9), indole and its derivatives (J10), succinimidyl active ester (A1, A6), carboxyl group (D1), primary amino group (C4), secondary amino group (C15), hydrazine group or substituted hydrazine group (C12), such as N-aminocarbazole, cyano group (G23), unsaturated bond of maleimide (E1), maleimide (C21), aldehyde group (D6), acrylate group (E2), methacrylate group (E3), oxime group (G24), and, And (3) carrying out coupling reaction on the hydroxyl (H1) and the functionalized polyethylene glycol to obtain the polyethylene glycol modified bio-related substance. The coupling reaction includes, but is not limited to, the coupling reaction described above. Among them, the starting materials of the category (J1-J10) include, but are not limited to, the fluorescent reagents disclosed above in the present invention.
2.3.1.11. Conversion to target functional groups based on reactive groups
The method can be realized by any one of the following modes: (1) direct modification, based on direct modification of reactive groups, to obtain target functional groups. By way of example, such as the conversion of a carboxyl group to an active ester, an active ester analog, an acid halide, a hydrazide, an ester, a thioester, a dithioester, such as the conversion of a hydroxyl group, a thiol group, an alkynyl group, an amino group, a carboxyl group, etc., to the corresponding protected structure, and the like. For example, the acid anhydride modifies a hydroxyl group, an amino group, or the like. (2) Coupling reaction between two reactive groups, using a heterofunctional reagent containing 1 reactive group and a target functional group as a raw material, and introducing the target functional group through the reaction between one of the reactive groups and the reactive group at the tail end of a polyethylene glycol chain. The reaction mode and method between two reactive groups are not particularly limited, and include, but are not limited to, the above-mentioned coupling reaction method, such as alkylation reaction, α, β -unsaturated bond addition reaction, alkynyl addition reaction, schiff base reaction combined reduction reaction, condensation reaction, azide-alkyne cycloaddition reaction, 1, 3-dipolar cycloaddition reaction, Diels-Alder reaction, thiol-yne reaction, thiol-ene reaction, thiol-vinyl reaction, condensation reaction, and the like. Among them, the alkylation reaction is preferably a reaction based on alkylation of a hydroxyl group, a mercapto group or an amino group, which in turn corresponds to formation of an ether bond, a thioether bond, a secondary amino group or a tertiary amino group. Wherein the condensation reaction includes, but is not limited to, condensation reactions that form ester groups, thioester groups, amide groups, imine linkages, hydrazone linkages, carbamate groups, and the like. For example, the target functional group is introduced by click reaction using a heterofunctionalizing agent containing groups such as azide, alkynyl, alkenyl, trithiocarbonate, mercapto, dienyl, furyl, 12,4, 5-tetrazinyl, cyanoxide, etc. and the target functional group as a raw material. The reaction between the two reactive groups is accompanied by the formation of a new bond, and typical examples of the newly formed divalent linking group include an amide bond, a urethane bond, an ester group, a secondary amine bond, a thioether bond, a triazole group and the like. (3) The target functional group is obtained by a combination of direct modification and coupling reaction.
2.3.2. Branched functionalization of polyethylene glycol chain ends
Branched functionalization refers to the introduction of a branching group at the end of a polyethylene glycol chain to attach a functional group. In this case, the number of functional groups at the end of the corresponding polyethylene glycol chain is more than 1. The polyethylene glycol chain ends to which the branching groups are introduced may be hydroxyl groups or linear functionalized reactive groups selected from class a-class H.
2.3.2.1. Terminal branching functionalization process
The functionalized modification process of the branched end comprises two links of introducing a branched group and introducing a functional group. The order of these two segments is not particularly limited. In this case, the terminal-branching functionalization can be achieved in several ways including, but not limited to: (1) the functionalized branched group directly reacts with the hydroxyl at the end of the polyethylene glycol chain; (2) carrying out functional modification on the terminal hydroxyl of the main chain polyethylene glycol, and then reacting with a functional branched group; (3) firstly introducing a branching group, and then carrying out functional modification on the branching group. Wherein the introduction of the branching groups may or may not form the linking group L 0 . Taking the terminal hydroxyl of polyethylene glycol as an example: when the branching group is attached by alkylation, the branching agent loses the leaving group and the hydroxyl group loses the hydrogen atom, at which time it is believed that no linking group is formed, or it is believed that a new linking group ether bond is formed, at which time L 0 Comprises CH 2 CH 2 O; for example, when the terminal hydroxyl group of polyethylene glycol reacts with a group such as isocyanate group or carboxyl group, the whole or part of NHCO, CO, etc. which is a new bond formed is contained in L 0 The preparation method comprises the following steps of (1) performing; as another example, succinic acid-functionalized polyethylene glycol termini can be reacted with a branching agent to form a linker containing a succinyl group. The method for the functional modification of the branched group is not particularly limited, and includes a functional modification based on a hydroxyl group, and also includes a functional modification based on a non-hydroxyl functional group to a new functional groupAnd (4) converting the clusters.
The method for introducing the above-mentioned branched group is not particularly limited, and the existing techniques in the chemical field may be adopted as long as a covalent bond can be formed, including but not limited to the various coupling reactions described above. By way of example, the preparation of comb structures is described in the literature { Macromolecules 2013, 46,3280-3287}, the literature { Macromolecules chem. Phys.2014,215,566-571}, the literature { Macromolecules,2012, 45,3039-3046}, the literature { Polymer chem.,2012,3,1714-1721}, US5,811,510, US7,790,150, US7,838,619, and the like, and the literature { Journal of Polymer Science, Part A: polymer Chemistry,2013,51, 995-. The branched structures described in the above-mentioned documents and the preparation thereof are incorporated herein by reference.
The method of functionalizing the terminal of the branched group is not particularly limited, and includes, but is not limited to, the above-mentioned linear functionalizing methods.
2.3.2.2. Terminal-branched functionalized starting materials
When end-difunctional, it is preferably selected from the group consisting of heterofunctional small molecules htriSM, aldehydes containing 1 epoxy group, alcohols containing 1 epoxy group (e.g. htriSM, htr, r, g, and htr, r, and t
Figure BDA0002963906360000641
) Sulfonate containing 1 epoxy group, halide containing 1 epoxy group, compound containing one epoxy group and 1 other reactive group. Also included are combinations of Michael addition reactions of primary amines with 2 molecules of acrylates. Or the lipoic acid is adopted to carry out end capping, and then the disulfide bond is reduced to open the ring, so that two sulfydryl groups at the tail end are obtained.
The heterofunctionalized small molecule htriSM includes but is not limited to the structures listed and cited in paragraphs [0902] to [0979] of the document CN 104877127A.
The heterofunctional small molecule htriSM contains two different functional groups, namely heterofunctional group pairs, wherein one functional group is 1, and the other functional group is two. The pairs of heterofunctional groups that can be present at the same time are described in section 2.2.2, and are not described in detail here.
The htriSM includes, but is not limited to, alcohols, thiols, primary amines, secondary amines, sulfonates or halides containing two naked or two protected hydroxyl groups (again, for example, triethanolamine p-toluenesulfonate, glycerol monothioglycolate, 3, 4-dihydroxy-2' -chloroacetophenone, and protected forms of the hydroxyl groups of htriSM described above), alcohols, thiols, primary amines, secondary amines, sulfonates or halides containing two or two protected thiol groups (again, for example, dimercaprol and protected forms of the thiol groups thereof), alcohols, thiols, primary amines, secondary amines, primary amines, or two protected secondary amines, sulfonates or halides, and the like. Among them, alcohols containing two primary amines are exemplified by 1, 3-diamino-2-propanol.
The htriSM also includes, but is not limited to: primary amines containing 2 hydroxyl groups, aldehydes containing 2 protected hydroxyl groups, aldehydes containing 1 epoxy group, primary amines containing 1 epoxy group, secondary amines containing 2 primary amino groups, sulfonic acids containing 2 hydroxyl groups, carboxylic acids containing 2 hydroxyl groups, azide compounds containing two hydroxyl groups, and protected forms of the hydroxyl groups described above. The primary amine containing 2 hydroxyl groups includes, but is not limited to, 2-amino-1, 3-propanediol, 2-amino-2-methyl-1, 3-propanediol, N-bis (2-hydroxyethyl) ethylenediamine, 3-amino-1, 2-propanediol, 2-amino-1- [4- (methylthio) phenyl ] glycol]1, 3-propanediol, 2-amino-1-phenyl-1, 3-propanediol, 2- (3, 4-dihydroxyphenyl) ethylamine, 2-amino-1, 3-benzenediol, and the like. The secondary amines containing 2 primary amino groups include, but are not limited to, diethylenetriamine, N- (3-aminopropyl) -1, 4-butanediamine, 3' -diaminodipropylamine, N- (2-aminoethyl) -1, 3-propanediamine, 3, 6-diaminocarbazole, and the like. The sulfonic acid containing two hydroxyl groups includes, but is not limited to, 6, 7-dihydroxynaphthalene-2-sulfonic acid, 1, 4-dihydroxyanthraquinone-2-sulfonic acid. The carboxylic acid having 2 hydroxyl groups (dihydroxymonocarboxylic acid) includes, but is not limited to, 2, 3-dihydroxypropionic acid,2, 2-dimethylolpropionic acid, 2, 4-dihydroxy-3, 3-dimethylbutyric acid, N-dihydroxyethylglycine, 2, 3-dihydroxybenzoic acid, 2, 4-dihydroxybenzoic acid, 2, 5-dihydroxybenzoic acid, 2, 6-dihydroxybenzoic acid, 3, 4-dihydroxybenzoic acid, 3, 5-dihydroxybenzoic acid, 3, 4-dihydroxycinnamic acid, 2, 6-dihydroxypyridine-4-carboxylic acid, 4, 8-dihydroxyquinoline-2-carboxylic acid. The azide compound containing two hydroxyl groups includes but is not limited to 3-azido-2, 3-dideoxy-1-O- (tert-butyldimethylsilyl) -beta-D-arabino-hexapyranose, 2-dimethylolpropionic acid azidohexyl ester. Wherein both hydroxy groups are protected, e.g. with a bishydroxy group, e.g.
Figure BDA0002963906360000651
The htriSM also includes, but is not limited to: 3-allyloxy-1, 2-propanediol, 5-norbornene-2, 3-dicarboxylic acid, 1-propynylglycerol ether, 2, 6-dihydroxy-3-cyano-4-methylpyridine, 1, 3-dibromo-2-propanol, 2, 3-dibromo-1-propanol, 1, 4-dibromo-2-butanol, 1, 4-diazido-2-butanol, 1, 3-dichloropropanol, 4' -dichlorobenzhydrol, 2-bromomalonaldehyde, 2-hydroxyhexanal, 2- (4-chlorophenyl) malonaldehyde, 2- (3-hydroxycarbonyl-6-pyridyl) malonaldehyde, 7-amino-1, 3-naphthalenedisulfonic acid, 4-chloro-1, 2-phenylenediamine, 4-bromoo-phenylenediamine, 6, 8-dimercaptooctanoic acid, 4-chloro-1, 3-benzenedithiol, 2, 6-bis (p-azidobenzylidene) -4-carboxycyclohexanone, hydroxydicarboxylic acids (including but not limited to tartronic acid, L-malic acid, D-malic acid, citramalic acid, 3-hydroxyglutaric acid), aminodicarboxylic acids (including but not limited to 2-aminomalonic acid, diethyl 2-aminomalonate, 3-aminoglutaric acid), mercaptodicarboxylic acids (including but not limited to mercaptosuccinic acid), 4-chlorophthalic acid, 2-bromosuccinic acid, itaconic acid, 4-amino-2- (2-aminoethylamino) butyric acid, succinic acid, malic acid, succinic acid, lactic acid, fumaric acid, maleic acid, fumaric acid, maleic acid, 4-amino-2- (2-aminoethylamino) butyric acid with two amino groups protected, glycerol dimethacrylate, 2-bis (allyloxymethyl) -1-butanol,
Figure BDA0002963906360000652
Etc. and the aboveAny of the functional groups in the number of 2 in protected form.
The htriSM also includes, but is not limited to: lysine, lysine with two amino groups protected, glutamic acid and aspartic acid.
Since both hydrogens in the primary amine can be substituted to form a trivalent N branching center, heterofunctionalized small molecules containing one primary amino group and another reactive group can also be used as htriSM. Examples thereof include diglycolamine, 2- (2-aminoethylmercapto) ethanol, 1-amino-2-propanol, 4-hydroxyphenylethylamine, mercaptoethylamine, N-methyl-1, 3-propanediamine, N-ethyl-1, 3-propanediamine, and N-isopropyl-1, 3-propanediamine.
When the terminal trifunctional is carried out, the method includes but is not limited to the use of a tetrafunctional small molecule htetraSM containing three hydroxyl groups and another reactive group, and includes but is not limited to: N-Trimethylol-2-aminoethanesulfonic acid, trimethylol-methylaminopropanesulfonic acid, methyl-6-O-p-toluenesulfonyl-alpha-D-glucoside, 2- (bromomethyl) -2- (hydroxymethyl) -1, 3-propanediol, tris, 2-amino-1, 3, 4-octadecanetriol, 3-aminopropylsilanetriol, 4- (2-amino-1-hydroxyethyl) -1, 2-benzenediol, 4- [ (2-isopropylamino-1-hydroxy) ethyl ] -1, 2-benzenediol, 3, 4-dihydroxy-alpha- ((methylamino) methyl) benzyl alcohol, 2, 5-anhydro-1-azido-1-deoxy-D-glucitol, 2,3, 4-trihydroxybutyraldehyde (L-erythrose, D-erythrose, L- (+) -threose, D- (+) -threose), 2,3, 4-trihydroxybenzaldehyde, 3,4, 5-trihydroxybenzaldehyde, tris (hydroxymethyl) methylglycine, 2,3, 4-trihydroxybutyric acid (including but not limited to erythronic acid, threonic acid), 2,4, 6-trihydroxybenzoic acid, shikimic acid, 3,4, 5-trihydroxybenzoic acid, 2,3, 4-trihydroxybenzoic acid, arjunolic acid, 1,4, 7-tri-tert-butoxycarbonyl-1, 4,7, 10-tetraazacyclododecane, tri-tert-butoxycarbonyl spermine, 1,4, 7-tri-tert-butoxycarbonyl-1, 4,7, 10-tetraazacyclododecane, and the like, and hydroxyl groups of any of the foregoing are protected. The compound may be selected from the group consisting of citric acid, agaricic acid, N-hydroxyethylethylenediaminetriacetic acid, pentaerythritol triacrylate, aminomethane tripropionic acid, and tri-tert-butyl aminomethane tripropionate. Also included are terminal branching reactions based on alkenyl, trichlorosilane and allylmagnesium chloride, referred to as Macromolecules, Vol.33, No.12,2000, to form tetravalent silicon-based branching centers. Also included are terminal branching reactions based on alkenyl, trichlorosilane, and allyl alcohol to form tetravalent siloxane branching centers. Also included are tri-functionalized small molecules such as 1,4, 7-tris (t-butoxycarbonylmethyl) -1,4,7, 10-azacyclotetradecane (NOTA), which require an excess of such tri-functionalized small molecules.
When the terminal tetrafunctionalization is carried out, the raw material may be selected from pentafunctionalized xylitol, 1, 5-anhydroglucitol, bis (2-hydroxyethyl) amino (trihydroxymethyl) methane, miglitol, D- (+) -talose, arbutin, diethylenetriaminepentaacetic acid, and the like. But preferably hetero-pentafunctionalized small molecules containing 2 functional groups. Including but not limited to 1,2,5, 6-diisopropylidene-alpha-D-isofuranose, 2,3,5, 6-di-O-cyclohexylidene-alpha-D-mannose, 2-azido-1, 3-bis [ (2, 2-dimethyl-1, 3-dioxan-5-yl) oxo ] propane, etc., containing four protected hydroxyl groups and one reactive group. But also molecules containing 2 epoxy groups and 1 reactive group. It may also be preferred to have a pentafunctional small molecule hpentSM with two functional groups, one in 4 and the other in 1: 2- (2-hydroxyethylamino) -2-hydroxymethyl-1, 3-propanediol, 2-hydroxymethylpiperidine-3, 4, 5-triol, 6-amino-4- (hydroxymethyl) -4-cyclohexyl- [4H,5H ] -1,2, 3-triol, fenoterol, benserazide, 1-azido-1-deoxy- β -D-galactopyranoside, 2-azidoethyl- β -D-glucopyranoside, 2,3,4, 5-tetrahydroxypentanal (including but not limited to ribose, arabinose, xylose, lyxose), 2,3,4, 5-tetrahydroxypentanoic acid (including but not limited to ribonic acid, arabinonic acid, xylonic acid, lyxonic acid), Diethylenetriamine, N- (3-aminopropyl) -1, 4-butanediamine, and the like, and 4-numbered functional groups of any of the above.
When the terminal penta-functionalization is performed, the starting material preferably contains two functional groups, one of which is a 5 number and the other of which is a 1 number hexafunctional small molecule hhexaSM, including but not limited to: sorbitol, mannitol, D-talitol, D-glucosamine, 1-mercaptosorbitol, N-methyl-D-glucosamine, 2,3,4,5, 6-pentahydroxyhexanal (including but not limited to beta-D-allose, D-altrose, D-anhydroglucose, D- (+) -mannose, L (-) -mannose, D-gulose, idose, D-galactose, L- (-) -talose, D- (+) -talose), 2,3,4,5, 6-pentahydroxyhexanoic acid (including but not limited to allose, altronic acid, gluconic acid, mannonic acid, gulonic acid, idonic acid, galactic acid, talonic acid), D-sorbitol-3-phosphate, and the like, and any of the above 5 number of functional groups are coated with a 5 number of functional groups A form of protection.
The starting material for the preparation of the dendritic structure may be selected from the group comprising, but not limited to, the following structures: htriSM, htetraSM, hpentSM, hhexaSM, an heterofunctionalized molecule containing 1 epoxy group and another reactive group, htriSM containing two ethynyl groups or a protected ethynyl group and another reactive group, diallyl (meth) silane, combinations of acrylates and diamines (repeating Michael addition reaction of primary amines to 2 molecules of acrylate, amidation reaction of ester groups), combinations of propargyl glycidyl ether with mercaptoethylamine, mercaptoethylamine hydrochloride, or amino-protected mercaptoethylamine (repeating addition reaction of primary amino groups to epoxy groups, click reaction of alkynyl groups to 2 mercapto groups), diallylmethylsilane, and the like. Specific examples are as follows
Figure BDA0002963906360000661
Epichlorohydrin, amino-protected lysine, glutamic acid, aspartic acid, N-dihydroxyethylglycine and hydroxy-protected forms thereof, dihydroxymonocarboxylic acids and hydroxy-protected forms thereof, hydroxydicarboxylic acids and hydroxy-protected forms thereof, aminodicarboxylic acids and amino-protected forms thereof, mercaptodicarboxylic acids and mercapto-protected forms thereof, glyceraldehyde and hydroxy-protected forms thereof, methyl-6-O-p-toluenesulfonyl-alpha-D-glucoside, 3-aminopropylsilanetriol, 2, 3, 4-trihydroxybutyraldehyde, 2, 3, 4-trihydroxybutyric acid, citric acid, N-hydroxyethylethylenediaminetriacetic acid, N-hydroxyiminocyanuric acid, N-hydroxycyanuric acid, N-2-amino-2, 3, 4-trihydroxybutyric acid, 4-lycine, N-di-hydroxyiminocyanuric acid, N-2-aminetriacetic acid, N-2-4-L-R-L-R-,
Figure BDA0002963906360000662
2-azido-1, 3-bis [ (2, 2-dimethyl-1, 3-dioxan-5-yl) oxo]Propane, and the like. Among them, 2-dimethylolpropionic acid is preferable as the dihydroxy monocarboxylic acid. The hydroxy dicarboxylic acid is preferably malic acid, 3-hydroxyGlutaric acid.
Monomers for preparing hyperbranched structures include, but are not limited to, monomers disclosed in the literature [ Journal of Polymer Science, Part A: Polymer Chemistry,2013,51, 995-: epoxy propanol, epoxy propanol,
Figure BDA0002963906360000663
(3-ethyl-3-oxetanylcarbinol),
Figure BDA0002963906360000664
Combinations of acrylates with diamines, and the like.
Monomers for preparing comb structures having repeating units include, but are not limited to: glycerol in which the 2-hydroxyl group is protected (forming a polyglycerol structure), pentaerythritol in which the two hydroxyl groups are protected (e.g., monophenylol pentaerythritol, forming polypentaerythritol), and mixtures thereof,
Figure BDA0002963906360000665
(F is as defined above, preferably in protected form, one of the preferred forms being protected hydroxy OPG 4 Such as 1-ethoxyethyl-2, 3-epoxypropyl ether, benzyl glycidyl ether, tert-butyl glycidyl ether, allyl glycidyl ether, propargyl glycidyl ether, glycidyl methacrylate, glycidyl ether, and a,
Figure BDA0002963906360000666
Figure BDA0002963906360000667
(e.g., azidopropyl methacrylate), carbon dioxide and
Figure BDA0002963906360000671
combinations of (e.g., { Macromolecules 2013,46, 3280-3287}, and carbon dioxide with a compound selected from the group consisting of
Figure BDA0002963906360000672
Propargyl glycidyl ether, and the like), diisocyanates, and forms containing 1 reactive group or protected forms thereofA combination of dihydric alcohols of formula,
Figure BDA0002963906360000673
Figure BDA0002963906360000674
In combination with diamines (forming a comb with a plurality of thiol groups suspended, { macromol. rapid commu.2014, 35,1986-1993}), D-glucopyranose units (forming acetalized glycan structures, such as poly (1 → 6) hexoses, 2, 1-polyfructose, in particular dextran and its oxidized structures, polyfructose structures as described in US5,811,510, US7,790,150, US7,838,619), lysine, aspartic acid, glutamic acid, etc. Other trihydric alcohols, trihydric alcohols containing one protected hydroxyl group, tetrahydric alcohols containing 2 protected hydroxyl groups, polyhydric alcohols containing 2 naked hydroxyl groups and other protected hydroxyl groups can be used as raw materials for preparing the comb-shaped structure. In addition, the comb-like structure may be a non-repeating structure, for example, a polypeptide structure formed by linking 2 or more amino acids selected from any 1 or more amino acids selected from lysine, aspartic acid, and glutamic acid with other amino acids (e.g., glycine) as a spacer. In addition, including but not limited to the above-mentioned 2,3,4, 5-tetrahydroxypentanal, 2,3,4, 5-tetrahydroxypentanoic acid, 2,3,4,5, 6-pentahydroxyhexanal, 2,3,4,5, 6-pentahydroxyhexanoic acid, D-glucosamine, 1-mercaptosorbitol, N-methyl-D-glucosamine, D-sorbitol-3-phosphate and the like can be directly used as a raw material for preparing the comb-like branched terminals.
Starting materials for the preparation of cyclic structures include, but are not limited to: 2, 5-anhydro-1-azido-1-deoxy-D-glucitol, 1,4, 7-tri-tert-butoxycarbonyl-1, 4,7, 10-tetraazacyclododecane, 2-hydroxymethylpiperidine-3, 4, 5-triol, 6-amino-4- (hydroxymethyl) -4-cyclohexyl- [4H,5H ] -1,2, 3-triol, 1-azido-1-deoxy-beta-D-galactopyranoside, 2-azidoethyl-beta-D-glucopyranoside, propargyl-alpha-D-mannopyranoside, propargyl-alpha-L-fucopyranoside, propargyl-beta-D-lactoside, beta-lactoside, and mixtures thereof, Monofunctional cyclodextrins (e.g., mono-6-O- (azido) - β -cyclodextrin, mono-6-O- (p-toluenesulfonyl) - γ -cyclodextrin, mono-2-O- (p-toluenesulfonyl) - γ -cyclodextrin, mono-6-O- (p-toluenesulfonyl) - β -cyclodextrin, mono-2-O- (p-toluenesulfonyl) - α -cyclodextrin), and the like.
2.3.2.3. Formation of the branching centres in the terminal G
The branching centers in the terminal G are each independently selected from the group consisting of, but not limited to, carbon atoms, nitrogen atoms, phosphorus atoms, silicon atoms, cyclic structures, combinations of any 2 or more of the foregoing. The trivalent branching center may be of a symmetrical structure or an asymmetrical structure.
The above-mentioned branching centers may be derived either directly from the starting materials or by coupling reactions between the starting materials.
As an example of being directly derived from the starting material, for example, the symmetrical trivalent carbon branching center may be derived from
Figure BDA0002963906360000675
Serinol, 2-dimethylolpropionic acid, etc., asymmetric trivalent carbon branching center can be derived from epichlorohydrin, 3-methylamino-1, 2-propanediol, malic acid, 3-hydroxyglutaric acid, lysine, glutamic acid, aspartic acid, etc., symmetric trivalent N branching center can be derived from N, N-bis (2-hydroxyethyl) ethylenediamine, N-dihydroxyethylglycine, etc., tetravalent carbon branching center can be derived from pentaerythritol, citric acid, etc., cyclic branching center can be derived from 3, 6-diaminocarbazole, 2, 5-anhydro-D-glucitol, methyl-D-mannoside, dihydroxybenzoic acid (including various isomers with different substitution positions), etc, Amino-benzenediol (including various isomers with different substitution positions),
Figure BDA0002963906360000676
The phosphorus atom branching center can be obtained from phosphoric acid, phosphoric acid ester, thiophosphoric acid ester as a raw material.
Branching centers obtained by coupling reactions between the starting materials. For example, alkylation or amidation of a secondary amine can give a trivalent nitrogen center. Such as primary amines with 2-molecule sulfonates, halides, epoxides, alpha, beta-unsaturation (e.g., acrylates), may also give a trivalent nitrogen branching center. As another example, reaction between an alkynyl group and 2 thiol groups can result in an asymmetric trivalent carbon branching center. As another example, a reaction between a B5 or B6-type functional group and a disulfide bond may form a trivalent carbon branching center. As another example, a tetravalent silicon branching center may be obtained by the above-described terminal branching reaction based on alkenyl, trichlorosilane, and allylmagnesium chloride, or based on alkenyl, trichlorosilane, and allyl alcohol. As another example, a trivalent silicon branching center can be obtained by reaction of diallylmethylsilane as a repeating unit. Asymmetric carbon-branched dimercapto can be obtained by reduction of the disulfide bond of the five-membered ring of the lipoic acid. By acetalization of p-hydroxymethylbenzaldehyde, a trivalent carbon branching center of an acetal structure can be obtained, and the branching center can be degraded. Also, reactive groups such as class B5 or class B6 react with disulfide bonds to give symmetric trivalent carbon branching centers.
2.4. Coupling reaction process
The scope of choice of the coupling reaction described in the present invention is not particularly limited, as long as two identical or different reactive groups can react to form a covalent linker. The reaction conditions, depending on the type of covalent linking group formed by the reaction, can be established by the techniques known in the art. Including but not limited to documents CN104877127A, CN104530413A, CN104530415A, CN104530417A and the references cited therein. For example, CN104530417A corresponds to segments [1212] to [1280], and CN104877127A corresponds to segments [0992] to [0997 ]. Including but not limited to any of the reactive groups disclosed in the present invention and the cited documents, can undergo a reaction that produces a covalent linking group. Other types of reactions mentioned in the present invention are also included. The valence state of the covalent linking group may be divalent or trivalent, with divalent predominance.
The coupling reaction can produce stable groups as well as degradable groups.
In general terms, for example: the amino group is respectively reacted with active ester, formic acid active ester, sulfonic ester, aldehyde, alpha, beta-unsaturated bond, carboxylic acid group, epoxide, isocyanate and isothiocyanate to obtain bivalent connecting group such as amido, urethane group, amino, imino (which can be further reduced into secondary amino), amino, amido, amino alcohol, urea bond, thiourea bond and the like (ii) a Reacting sulfhydryl with divalent linking groups containing active ester, formic acid active ester, sulfonic ester, sulfhydryl, maleimide, aldehyde, alpha, beta-unsaturated bond, carboxylic acid group, iodoacetamide and acid anhydride to obtain thioester group, thiocarbonate, thioether, disulfide, thioether, thio hemiacetal, thioether, thioester, thioether and imide; unsaturated bonds react with sulfydryl to obtain thioether groups; carboxyl or acyl halide reacts with sulfhydryl and amino respectively to obtain thioester group, amide group and other groups; hydroxyl reacts with carboxyl, isocyanate, epoxide and chloroformyl to obtain divalent connecting groups such as ester group, carbamate group, ether bond, carbonate group and the like; reacting carbonyl or aldehyde group with amino, hydrazine and hydrazide to obtain divalent connecting groups such as imine bond, hydrazone, acylhydrazone and the like; reactive groups such as azide, alkynyl, alkenyl, sulfydryl, azide, diene, maleimide, 1,2, 4-triazoline-3, 5-diketone, dithioester, hydroxylamine, hydrazide, acrylate, allyloxy, isocyanate, tetrazole and the like are subjected to click chemistry reaction to generate corresponding divalent connecting groups containing structures such as triazole, isoxazole, thioether bonds and the like. The types of click reactions and resulting linkers reported in and cited in the document adv.funct.mater, 2014,24,2572 are incorporated herein by reference, specifically azide-alkynyl cycloaddition, Diels-Alder addition, oxime or acylhydrazone production, mercapto-vinyl addition, mercapto-alkynyl addition, mercapto-isocyanate group, 1, 3-dipolar cycloaddition, and the like. Also included, but not limited to, cycloaddition reactions, Diels-Alder addition reactions, 1, 3-dipolar cycloaddition reactions, and the like, which can occur in the following class G. The primary amine reacts with one molecule of sulfonate, halide, epoxide and alpha, beta-unsaturated bond to obtain divalent secondary amino group, and reacts with two molecules to form trivalent tertiary amino group. Also as in the reaction between a B5 or B6-type functional group and a disulfide bond of the present invention, a trivalent linking group can be formed. As another example, the reactive group E13 reacts with a disulfide bond to form a trivalent linking group
Figure BDA0002963906360000681
Also as follows for the reaction of hydrazine and aldehyde groups
Figure BDA0002963906360000682
Typical examples of the divalent linking group to be formed include an amide bond, a urethane bond, an ester group, a secondary amine bond, a thioether bond, a triazole group and the like. When an amide bond (-CONH-) or an imide (-CON (-) 2 ) When used, the synthesis can be carried out in ways including, but not limited to: (1) obtained by condensation reaction between amino and carboxyl; (2) obtained by reaction between an amino group and a carboxylic acid derivative; and (3) the reaction is realized by amidation reaction of substrate amine and acyl halide, wherein the acyl halide is preferably acyl chloride. When a urethane linkage (-OCONH-) is formed, the compound can be obtained by condensation reaction of a terminal amino group and a terminal active carbonate derivative; wherein the active formate can be a derivative which can react with amino to obtain a urethane bond, and the derivative comprises but is not limited to Succinimidyl Carbonate (SC), p-nitrophenol carbonate (p-NPC), 2,4, 6-trichlorophenol carbonate, imidazole carbonate, N-hydroxybenzotriazole carbonate, preferably Succinimidyl Carbonate (SC), o-nitrophenol carbonate (o-NPC) and the like; urethane linkages can also be obtained by reacting hydroxyl groups with isocyanates. When a monothio or dithio carbamate linkage is formed, it can be obtained by reacting a terminal amino group with a terminal thiooxycarbonylchloride, reacting a hydroxyl or mercapto group with an isothiocyanate, or reacting a mercapto group with an isocyanate. When an ester bond (-OCO-) is formed, it can be obtained by a condensation reaction of a terminal hydroxyl group with a terminal carboxyl group or an acid halide, preferably an acid chloride. When a secondary amine linkage (-CH) is formed 2 NHCH 2 -) can be obtained by condensation and reduction reaction between aldehyde group and amino group, or by alkylation reaction between primary amine and sulfonic ester or halide. When a thioether bond is formed: (>CHS-) can be obtained by addition reaction between a terminal mercapto group and maleimide or other reactive group containing an unsaturated bond ({ Angew. chem. int. Ed., 2010,49,3415-3417}), or by alkylation reaction between a terminal mercapto group and a sulfonate or halide. When a triazole group is formed, it can be obtained by a click reaction between an alkynyl group and an azide. When 4, 5-dihydroisoxazoles are formed, this may be by reaction between the cyanide and the alkynylCarrying out 1, 3-dipolar cycloaddition reaction.
Typical reactions to form stable divalent linking groups are of the alkylation type, including but not limited to the alkylation of hydroxyl, mercapto or amino groups with sulfonate esters or halides, which in turn correspond to the formation of ether linkages, thioether linkages, secondary or tertiary amino groups.
2.5. Purification and characterization of intermediates and products
The intermediates or products prepared in the present invention can be purified by purification methods including, but not limited to, extraction, recrystallization, adsorption treatment, precipitation, reverse precipitation, membrane dialysis, or supercritical extraction. The structure, molecular weight and molecular weight distribution of key intermediates and products are characterized and confirmed by methods including but not limited to nuclear magnetism, electrophoresis, ultraviolet-visible spectrophotometer, FTIR, AFM, GPC, HPLC, MALDI-TOF, circular dichroism method and the like. For monodisperse nitrogen-branched nine-arm polyethylene glycol derivatives, the molecular weight is preferably confirmed by MALDI-TOF. Methods for determining the attribution of characteristic peaks in nuclear magnetic tests include, but are not limited to, those described and recited in documents CN104877127A, CN104530413A, CN104530415A, CN104530417A, and the respective cited documents. The terminal functionalization rate (substitution rate) of the nitrogen-branched nine-arm polyethylene glycol derivative is the mol percentage of the terminal hydroxyl group of the nitrogen-branched nine-arm polyethylene glycol which is functionalized and modified, and the terminal hydroxyl group characteristic peak-CH in the nuclear magnetism characteristic peak of the nitrogen-branched nine-arm polyethylene glycol raw material is obtained 2 CH 2 OH and EO Block characteristic Peak-CH 2 CH 2 Characteristic peak of functional group and characteristic peak of EO block-CH in nuclear magnetic characteristic peak of O-, and nitrogen branched nine-arm polyethylene glycol derivative product 2 CH 2 The integral ratio of O-is obtained by conversion, and the conversion method is well known to those skilled in the art and will not be described herein.
2.6. Molecular weight and PDI determination
The measurement is carried out by GPC, MALDI-TOF or the like. The molecular weight deviation is controlled within 10 percent, and can reach within 8 percent in some cases, even can reach within 5 percent. PDI is controlled to be less than 1.10-1.08, and for most molecular weights of 5-40 kDa, the molecular weight is stably controlled to be less than 1.05, and part of the molecular weight is less than 1.03, and can be less than 1.02, and the best is about 1.01.
3. The invention also provides a nitrogen-branched nine-arm polyethylene glycol derivative modified biologically-relevant substance, which has the following structural general formula:
Figure BDA0002963906360000691
wherein N is a nitrogen atom trivalent branching center;
U 0 being a tetravalent radical, 3U's in the same molecule 0 The same; n and three U 0 Together form a nine-valent branched structure;
L 2 is absent or L 2 Is a divalent linking group; in the same molecule, nine L 2 Identical or different, preferably identical, from one another;
PEG is a polyethylene glycol block taking ethylene oxide as a repeating unit, and both ends of the PEG are oxygen radicals;
k is the number of EF in a single functionalized end and is selected from 1 or 2-250;
in a single functionalized PEG chain, g is 0 or 1; g is a terminal branching group selected from a trivalent or higher valent connecting group which connects the PEG chain segment with a terminal functional group; l is 0 Is a divalent linking group, connecting the PEG chain segment with the terminal branching group G;
when g is 0, k is 1, L 0 G does not exist;
when G is 1, G is present, L 0 May be present or absent, k is 2-250;
EF may be expressed as ED (structure is
Figure BDA0002963906360000701
) Or EF 1 (structure is
Figure BDA0002963906360000702
) And D is not equal to E 01 (ii) a Wherein q and q are 1 Each independently is 0 or 1; z 1 、Z 2 Each independently is a divalent linking group; wherein D is modified bio-related substance and nitrogen branched nine-arm polyethyleneResidues formed after the reaction of the diol derivative; e 01 Is selected from R 01 Protected R 01 Deprotected R 01 Or blocked R 01 ；R 01 Is a reactive group capable of reacting with a biologically relevant substance; l is a linking group formed after a reactive group in the nitrogen branched nine-arm polyethylene glycol derivative reacts with a biologically relevant substance; wherein the number of D at the end of one branch chain is marked by k D ，0≤k D K is not more than k, k of each branched chain in the same molecule D Each independently the same or different, and the sum of the numbers of D (N) in any one of the nitrogen-branched nine-arm polyethylene glycol derivative molecules D ) At least 1, preferably at least 9; when G is 1, then G- (EF) k Can be expressed as
Figure BDA0002963906360000703
The biologically-relevant substance preferably has a plurality of reaction sites, and the same biologically-relevant substance and the same R 01 The residues D obtained by the reaction may be the same or different;
the biologically-relevant substance modified by the nitrogen-branched nine-arm polyethylene glycol derivative can exist stably or can be degraded; in the same molecule, U 0 、L 2 、L 0 、G、(Z 2 ) q -(Z 1 ) q1 、(Z 2 ) q Any of L, or any of the linkages formed with adjacent groups, may each independently be stable or degradable.
In the general formula (2), when g is 0, the structure of the bio-related substance modified by the nitrogen-branched nine-arm polyethylene glycol derivative is represented by the formula (3).
Figure BDA0002963906360000704
For the aforementioned general formula (2), when g is 1, the D content is 75%, or more than 75%, or less than 75%; preferably greater than 80%, more preferably greater than 85%, more preferably greater than 90%, more preferably greater than 94%, and most preferably equal to 100%. Wherein, when the g is 1 and the D content is 100 percent, the structure of the biologically-relevant substance modified by the nitrogen-branched nine-arm polyethylene glycol derivative is shown as a formula (4).
Figure BDA0002963906360000705
K at the end of nine PEG chains in the same molecule D Preferably all satisfy 1. ltoreq. k D K, i.e.at least one D is attached to each branch chain. Ideally, k is at the end of nine PEG chains in the same molecule D All satisfy k D K, i.e. all terminal reaction sites are each independently linked to a D, the percentage of D reaching 100%.
The D is from the same biologically relevant substance, but allows for different reaction sites with R 01 The residue formed after the reaction. Especially when there are multiple identical reactive groups in the biologically relevant material.
The D content (the number of D relative to the percentage of the reaction sites) in a single molecule is not particularly limited and may be greater than about 75%, equal to about 75%, or less than about 75%. The macroscopic material constituting the product of modification with a nitrogen-branched nine-arm polyethylene glycol derivative may have the same or different D content in each molecule, such as equal to 100%, or such as between 65% and 90%, or such as between 75% and 94%, for example. The higher the content of D, that is, the higher the drug loading rate, the more easily the effect of the biologically relevant substances is improved, and simultaneously, the higher the uniformity of the product structure is, the better the performance is. When there are a plurality of reaction sites in the bio-related substance, the same bio-related substance is used with the same R 01 After the reaction, the same or different residues D can be obtained, preferably the same residues D are obtained, in which case the more uniform and stable the properties of the product are. Preferably, the D content in a single molecule is greater than about 75%, more preferably greater than about 80%, more preferably greater than about 85%, more preferably greater than about 90%, more preferably greater than about 94%, and most preferably equal to 100%. For macroscopic materials, the average content of D may be greater than about 75% or less than about 75%, preferably greater than about 75%, more preferably greater than about 80%, more preferably greater than about 85%, more preferably greater than about 90%, more preferably greater than about 94%, and most preferably equal to 100%.
Wherein k is D The number of sites actually reacting with biologically-relevant substances in functional groups in a single molecule is expressed as a mean value for macroscopic substances, namely the number of reaction sites in an average nitrogen-branched nine-arm polyethylene glycol derivative molecule can be an integer or a non-integer; wherein the integer k in a single molecule D Each independently 0, 1 or 2 to 250. The invention also covers the substances of combining one molecule of the biological related substance with 2 or more molecules of the nitrogen-branched nine-arm polyethylene glycol derivative, but preferably 1 molecule of the biological related substance reacts with only 1 functional group, namely only one molecule of the nitrogen-branched nine-arm polyethylene glycol derivative is connected, and the corresponding quality controllability is strong. I.e. k D Also, the number of the bio-related substance molecules bound in F is shown (as a mean value for the macro substance, i.e., the number of bio-related substances to which one nitrogen-branched nine-arm polyethylene glycol derivative molecule is linked on average). The functional group modified by the nitrogen-branched nine-arm polyethylene glycol derivative can be totally or partially involved in modification of biological related substances. Preferably all participate in the modification of the biologically relevant substance. In the nitrogen-branched nine-arm polyethylene glycol derivative modified biological related substances, functional groups which are not combined with the biological related substances can be in a structural form before reaction, a protected form, a deprotected form or a non-biological related substance end capping form.
L may be covalently or non-covalently attached. Preferably a covalent linker; it may also be a double or multiple hydrogen bond. The nine PEG chain ends of the same molecule are allowed to correspond to different L, preferably the L of the nine PEG chain ends are the same, since reactions with different sites from the same biologically relevant substance are allowed. Any one of L is independently stable or degradable, and the connecting group of L and the adjacent heteroatom group is stable or degradable, and the condition for distinguishing stable existence and degradation is selected from any one of the following conditions: light, heat, enzymes, redox, acidic, basic, physiological conditions or in vitro simulated environments. Accordingly, any one of (Z) 2 ) q Each L is independently stably present or degradable, and (Z) 2 ) q The linking group of-L to the adjacent heteroatom group may be stable or degradable. Preferably, the L's at the nine PEG chain ends have the same stability, i.e., are all stably present or all degradable, in which case (Z) at the nine PEG chain ends 2 ) q -L also has the same stability.
The difference of the stability (also called degradability) of the biologically relevant substance modified by the nitrogen-branched nine-arm polyethylene glycol derivative according to the degradation position includes but is not limited to the following cases:
(1) g-0, having stable nine-valent centers
Figure BDA0002963906360000711
stabilized-O- (Z) 2 ) q -L-；
(2) g-0, having stable nine-valent centers
Figure BDA0002963906360000712
degradable-O- (Z) 2 ) q -L-；
(3) g 0, having a degradable nine-valent center
Figure BDA0002963906360000713
degradable-O- (Z) 2 ) q -L-；
(4) g 1, having a stable nine-valent center
Figure BDA0002963906360000714
stabilized-O-L 0 -G- (G and Z are excluded) 2 Connection of), stable- (Z) 2 ) q -L- (containing G and Z) 2 The connection of (c);
(5) g 1, having a stable nine-valent center
Figure BDA0002963906360000715
degradable-O-L 0 -G- (G and Z are excluded) 2 Connection of (c), stable- (Z) 2 ) q -L- (containing G and Z) 2 The connection of (c);
(6) g 1, having a stable nine-valent center
Figure BDA0002963906360000716
stabilized-O-L 0 -G- (G and Z are excluded) 2 Linked) degradable- (Z) 2 ) q -L- (containing G and Z) 2 The connection of (c);
(7) g 1, having a degradable nine-valent center
Figure BDA0002963906360000721
stabilized-O-L 0 -G- (G and Z are excluded) 2 Connection of (c), stable- (Z) 2 ) q -L- (containing G and Z) 2 The connection of (c);
(8) g-1 with degradable nine-valent centers
Figure BDA0002963906360000722
degradable-O-L 0 -G- (not containing G and Z) 2 Connection of), stable- (Z) 2 ) q -L- (containing G and Z) 2 The connection of (c);
(9) g-1 with degradable nine-valent centers
Figure BDA0002963906360000723
stabilized-O-L 0 -G- (not containing G and Z) 2 Ligation), degradable- (Z) 2 ) q -L- (containing G and Z) 2 The connection of (c);
(10) g 1, having a stable nine-valent center
Figure BDA0002963906360000724
degradable-O-L 0 -G- (not containing G and Z) 2 Linked) degradable- (Z) 2 ) q -L- (containing G and Z) 2 The connection of (c);
(11) g 1, having a degradable nine-valent center
Figure BDA0002963906360000725
degradable-O-L 0 -G- (G and Z are excluded) 2 Linked) degradable- (Z) 2 ) q -L- (containing G and Z) 2 The connection of (c);
(12) g-0, having stable nine-valent centers
Figure BDA0002963906360000726
stabilized-O- (Z) 2 ) q -, degradable L;
(13) g 1, having a stable nine-valent center
Figure BDA0002963906360000727
stabilized-O-L 0 -G-[(Z 2 ) q -] k A degradable L;
(14) g-0, having stable nine-valent centers
Figure BDA0002963906360000728
stabilized-O- (Z) 2 ) q Degradable L-D;
(15) g 1, having a stable nine-valent center
Figure BDA0002963906360000729
stabilized-O-L 0 -G-[(Z 2 ) q -] k Degradable L-D.
The combination of the different degradation amounts and degradation positions has been described above and will not be described further here. Wherein, (1), (4), (12) and (13) correspond to stable nitrogen branched nine-arm polyethylene glycol derivatives; (3) degradable nitrogen-branched nine-arm polyethylene glycol derivatives corresponding to the items (5), (7) to (11); (2) provided that (Z) in (1) and (6) 2 ) q - (containing Z) 2 The linkage with the PEG terminal O or G) and L can be degraded, so that the degradable nitrogen-branched nine-arm polyethylene glycol derivative can be used as the degradable stable nitrogen-branched nine-arm polyethylene glycol derivative containing L and can also be used as the degradable nitrogen-branched nine-arm polyethylene glycol derivative containing L. One preference of the above combination is to have stable nine-valent centers
Figure BDA00029639063600007210
Comprises (1), (2), (4), (5), (6), (10), (12), (13), (14) and (15). Wherein, in the two cases (1) and (4), the PEG part is not degradable, and the connection with the bio-related substances is also realizedThe biologically-relevant substance stably exists, and the biologically-relevant substance modified by the polyethylene glycol derivative can be a modified biologically-relevant substance; when the PEG is a basic drug, if other spacers are not connected between the basic drug and the PEG to generate degradable groups, the PEG can be metabolized as a whole, and the PEG has the advantages of accelerating solubilization, improving the interaction rate of the drug and a focus point or target tissue, improving the treatment effect and the like. For (14) and (15), D and (Z) may be passed 1 ) q1 -R 01 Can produce degradable L, and can also produce stable L: when D itself is not degradable, L may be degradable; when D itself is degradable, wherein the base drug and the spacer contain a degradable ester group, then L is preferably stably present, or a more stable degradable group (e.g., a more stable urethane linkage than an ester group) is preferred.
The nine PEG chains in the general formula (2) are from the same source, so the lengths of the nine PEG chains are completely the same or close to each other, and the biologically-relevant substances modified by the nitrogen-branched nine-arm polyethylene glycol derivative are more uniform in components, better in controllability and more stable in performance, and are more suitable for practical application and large-scale production. The PEG chain length can be n 1 ＝n 2 ＝n 3 ＝n 4 ＝n 5 ＝n 6 ＝n 7 ＝n 8 ＝n 9 N corresponds to a monodisperse PEG chain, or n 1 ≈n 2 ≈n 3 ≈n 4 ≈n 5 ≈n 6 ≈n 7 ≈n 8 ≈n 9 N corresponds to the polydisperse PEG chain.
3.1. Biologically relevant substances
The bio-related substance of the general formula (2) may be a bio-related substance, a modified bio-related substance, or a complex bio-related substance. Wherein the compound bio-related substances are chemical conjugates of 2 or more than 2 bio-related substances.
The bio-related substance may be a naturally occurring bio-related substance or an artificially synthesized bio-related substance. The biologically-relevant substance is obtained in a manner not particularly limited, and includes, but is not limited to, natural extracts and derivatives thereof, degradation products of natural extracts, gene recombination products (molecular cloning products), chemically synthesized substances, and the like. The hydrophilicity and hydrophobicity of the bio-related substance is not particularly limited, and may be hydrophilic or water-soluble, or may be hydrophobic or fat-soluble. The charge property of the bio-related substance is not particularly limited.
The biologically relevant substance may be the biologically relevant substance itself, or may be a dimer or multimer, partial subunit or fragment thereof, or the like.
The biologically-relevant substance may be a biologically-relevant substance itself, or a precursor, an activated state, a derivative, an isomer, a mutant, an analog, a mimetic, a polymorph, a pharmaceutically-acceptable salt, a fusion protein, a chemically-modified substance, a gene recombinant substance, or the like thereof, or a corresponding agonist, activator, inhibitor, antagonist, modulator, receptor, ligand or ligand, an antibody or a fragment thereof, an acting enzyme (e.g., a kinase, a hydrolase, a lyase, an oxidoreductase, an isomerase, a transferase, a deaminase, a deiminase, a invertase, a synthetase, or the like), a substrate for an enzyme (e.g., a substrate for a coagulation cascade protease, or the like), or the like. The derivatives include, but are not limited to, glycosides, nucleosides, amino acids, and polypeptide derivatives. Chemical modification products of new reactive groups are formed, and modification products generated after structures such as functional groups, reactive groups, amino acids or amino acid derivatives, polypeptides and the like are additionally introduced belong to chemical modification substances of biological related substances. The biologically-relevant substance may also allow for a target molecule, adjunct or delivery vehicle to be bound thereto, either before or after binding to the nitrogen-branched nine-armed polyethylene glycol derivative, to form a modified biologically-relevant substance or a complexed biologically-relevant substance. Wherein, the pharmaceutically acceptable salt can be inorganic salt, such as hydrochloride, or organic salt, such as oxalate, malate, citrate, etc.
The source of the biologically-relevant substance is not particularly limited and includes, but is not limited to, human, rabbit, mouse, sheep, cow, pig, and the like.
The application fields of the above biologically-relevant substances are not particularly limited, and include, but are not limited to, medical, regenerative medicine, tissue engineering, stem cell engineering, bioengineering, genetic engineering, polymer engineering, surface engineering, nano engineering, detection and diagnosis, chemical staining, fluorescent labeling, cosmetics, foods, food additives, nutrients, and the like. The biologically relevant substances in medicine, including but not limited to drugs, drug carriers and medical devices, can be used in various aspects such as disease treatment and prevention, wound treatment, tissue repair and replacement, image diagnosis and the like. By way of example, the related substances may also include: dye molecules for quantitative or semi-quantitative analysis; fluorocarbon molecules and the like which are useful for imaging diagnosis, blood substitutes, and the like; for example, antiparasitic agents such as primaquine and the like; for example, as a carrier for antidotes such as the chelating agents ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), and the like. When the bio-related substance is used as a drug, the therapeutic field thereof is not particularly limited, and includes, but is not limited to, drugs for treating cancer, tumor, liver disease, hepatitis, diabetes, gout, rheumatism, rheumatoid, senile dementia, cardiovascular diseases and the like, antiallergic drugs, anti-infective agents, antibiotic agents, antiviral agents, antifungal agents, vaccines, central nervous system inhibitors, central nervous system stimulants, psychotropic drugs, respiratory tract drugs, peripheral nervous system drugs, drugs acting at synaptic or neuroeffector junction sites, smooth muscle active drugs, histaminergic agents, antihistaminicergic agents, blood and hematopoietic system drugs, gastrointestinal tract drugs, steroid agents, cell growth inhibitors, anthelmintic agents, antimalarial agents, antiprotozoal agents, antimicrobial agents, anti-inflammatory agents, immunosuppressive agents, drugs, anti-inflammatory agents, drugs, anti-inflammatory drugs, and drugs, anti-inflammatory drugs, and drugs, alzheimer's disease drug or compound, imaging agent, antidote, antispasmodic, muscle relaxant, anti-inflammatory agent, appetite suppressant, migraine-treating agent, muscle contractant, antimalarial, antiemetic/antiemetic, bronchodilator, antithrombotic, antihypertensive, antiarrhythmic, antioxidant, antiasthmatic, diuretic, lipid-regulating agent, antiandrogenic agent, antiparasitic, anticoagulant, antineoplastic agent, hypoglycemic agent, nutritional agent, additive, growth supplement, anti-inflammatory agent, vaccine, antibody, diagnostic agent (including but not limited to contrast agent), contrast agent, hypnotic, sedative, psychostimulant, tranquilizer, anti-parkinson's disease agent, analgesic, anxiolytic agent, myoinfective, auditory disorder agent, and the like. Wherein typical anti-cancer or anti-tumor drugs include, but are not limited to, breast cancer, ovarian cancer, cervical cancer, uterine cancer, endometrial cancer, gastrointestinal cancer, intestinal cancer, metastatic large intestine cancer, rectal cancer, colon cancer, colorectal cancer, gastric cancer, squamous cell cancer, laryngeal cancer, esophageal cancer, carcinoma, lung cancer, small cell lung cancer (small cell lung cancer), non-small cell lung cancer, liver cancer, thyroid cancer, kidney cancer, bile duct cancer, brain cancer, skin cancer, pancreatic cancer, prostate cancer, bladder cancer, testicular cancer, nasopharyngeal cancer, head and neck cancer, gallbladder and bile duct cancer, retinal cancer, renal cell cancer, gallbladder adenocarcinoma, multidrug resistant cancer, melanoma, lymphoma, non-Hodgkin's lymphoma, adenoma, leukemia, chronic lymphocytic leukemia, multiple myeloma, brain tumor, Wilms's tumor, liposarcoma, endometrial sarcoma, and colon cancer, Rhabdomyosarcoma, neuroblastoma, AIDS-related cancer (such as Kaposi's sarcoma), primary or secondary carcinoma, sarcoma or carcinosarcoma.
"drug" in the context of the present invention includes any agent, compound, composition or mixture that provides a physiological or pharmacological effect, either in vivo or in vitro, and often provides a beneficial effect. The class is not particularly limited and includes, but is not limited to, pharmaceuticals, vaccines, antibodies, vitamins, foods, food additives, nutritional agents, nutraceuticals, and other agents that provide a beneficial effect. The "drug" is not particularly limited in the range that produces physiological or pharmacological effects in vivo, and may be a systemic effect or a local effect. The activity of the "drug" is not particularly limited, and is mainly an active substance that can interact with other substances, and may also be an inert substance that does not interact with other substances; however, inert drugs can be converted to the active form by in vivo action or some stimulus.
The species of the bio-related substance is not particularly limited, and includes, but is not limited to, the following: drugs, proteins, polypeptides, oligopeptides, protein mimetics, fragments and analogs, enzymes, antigens, antibodies and fragments thereof, receptors, small molecule drugs, nucleosides, nucleotides, oligonucleotides, antisense oligonucleotides, polynucleotides, nucleic acids, aptamers, polysaccharides, proteoglycans, glycoproteins, steroids, lipid compounds, hormones, vitamins, vesicles, liposomes, phospholipids, glycolipids, dyes, fluorescent substances, targeting factors, cytokines, neurotransmitters, extracellular matrix materials, plant or animal extracts, viruses, vaccines, cells, micelles, and the like.
3.1.1. Classification and enumeration of biologically-relevant substances
The biologically relevant substances are classified and listed below. A biologically relevant substance may be present in one or more of the following categories. In general terms, the biologically relevant substances of general formula (2) of the present invention include, but are not limited to, those described and exemplified in documents CN104877127A, CN104530413A, CN104530415A and CN 104530417A. CN104530413A includes paragraphs [0813] to [0921], paragraphs [0971] to [1146], examples, paragraphs [0827] to [0870] and paragraphs [0889] to [0901] of CN104877127A, which are not described herein again.
3.2. Linking group L for connecting biologically relevant substance and polyethylene glycol branched chain
The structure of the covalent bond linking group L formed after the functional group in the nitrogen-branched nine-arm polyethylene glycol derivative reacts with the reactive group in the bio-related substance is related to the reactive groups of the bio-related substance and the polyethylene glycol. Including but not limited to documents CN104877127A, CN104530413A, CN104530415A, CN 104530417A. For example, CN104530413A corresponds to paragraphs [0922] to [0935] and the examples section.
In general terms, the use of a single,
the reaction site in the bio-related substance is not particularly limited, and may be a naturally occurring reaction site, or a modified activated group or an introduced reactive group. For example, in the case of drug molecules, common naturally occurring reactive sites are amino, thiol, carboxyl, disulfide, N-amino, C-carboxyl, hydroxyl (alcoholic hydroxyl, phenolic hydroxyl, etc.), carbonyl, guanidino, and the like. Document { Journal of Con the reactive sites for the amino acids described in trolled Release,161(2012): 461-. Non-naturally occurring groups, modified to introduce reactive sites including, but not limited to, any of R in classes A through H as described above 01 Examples thereof include aldehyde group, alkynyl group, azide group and the like.
The reactive group in the bio-related substance includes, but is not limited to, any one of an amino group, a thiol group, a disulfide group, a carboxyl group, a hydroxyl group, a carbonyl group or an aldehyde group, an unsaturated bond, and an introduced reactive group. For example: respectively reacting the amino-containing biologically-relevant substances with polyethylene glycol containing active ester, formic acid active ester, sulfonate, aldehyde, alpha, beta-unsaturated bonds, carboxylic acid groups, epoxide, isocyanate and isothiocyanate to obtain polyethylene glycol modifiers connected with groups such as amide groups, urethane groups, amino groups, imino groups (which can be further reduced into secondary amino groups), amino groups, amide groups, amino alcohols, urea bonds, thiourea bonds and the like; respectively reacting a biological related substance containing sulfydryl with polyethylene glycol containing active ester, formic acid active ester, sulfonic ester, sulfydryl, maleimide, aldehyde, alpha, beta-unsaturated bonds, carboxylic acid groups, iodoacetamide and anhydride to obtain a polyethylene glycol modifier connected with groups such as thioester, thiocarbonate, thioether, disulfide, thioether, thio hemiacetal, thioether, thioester, thioether, imide and the like; reacting biologically-relevant substances containing unsaturated bonds with polyethylene glycol containing sulfydryl to obtain a polyethylene glycol modifier connected with a thioether group; respectively reacting biologically-relevant substances containing carboxylic acid with polyethylene glycol containing sulfydryl and amino to obtain polyethylene glycol modifiers connected with thioester groups, amide groups and other groups; respectively reacting biologically-relevant substances containing hydroxyl with polyethylene glycol containing carboxyl, isocyanate, epoxide and chloroformyl to obtain polyethylene glycol modifier with ester group, carbamate group, ether bond, carbonate group and other groups; biologically relevant substances containing carbonyl or aldehyde groups respectively react with polyethylene glycol containing amino, hydrazine and hydrazide to obtain polyethylene glycol modifiers with imine bonds, hydrazone, acylhydrazone and other groups; reactive groups containing azide, alkynyl, alkenyl, sulfydryl, azide, diene, maleimide, 1,2, 4-triazoline-3, 5-diketone, dithioester, hydroxylamine, hydrazide, acrylate, allyloxy, isocyanate, tetrazole and the like are subjected to click chemistry reaction to generate various connecting groups containing structures such as triazole, isoxazole, thioether bonds and the like. Linkers produced by the click reaction reported in the document adv. funct. mater, 2014,24,2572 and cited therein are incorporated herein by reference.
The structure of L is not particularly limited, and includes, but is not limited to, a linear structure, a branched structure, or a cyclic-containing structure.
The valence of L is not particularly limited, and may be, for example, a divalent linking group, or a trivalent or higher covalent linking group. L is preferably a divalent linking group. Generally, a divalent linking group is formed. Trivalent linkers, such as those formed by reacting a thiol group with an alkynyl group, are exemplified. As another example, a reactive group of the type B5 can be reacted with a disulfide bond to provide a trivalent linking group
Figure BDA0002963906360000751
The stability of L is not particularly limited, and may be a linker that can exist stably or a degradable linker. The conditions which can be stably present, degradable conditions are in accordance with the term moiety. The L is preferably a linker that is stable under light, heat, low temperature, enzymatic, redox, acidic, basic, physiological conditions or in vitro simulated environments, or a linker that is degradable under light, heat, low temperature, enzymatic, redox, acidic, basic, physiological conditions or in vitro simulated environments. More preferably, L is a linker that is stable under light, heat, low temperature, enzyme, redox, acidic or basic conditions, or is a linker that is degradable under light, heat, low temperature, enzyme, redox, acidic or basic conditions.
When a linker group that can be stably present, L can contain a linker group including, but not limited to, an ether linkage, a thioether linkage, a urea linkage, a thiourea linkage, a carbamate group, a thiocarbamate group, a secondary amino group, a tertiary amino group, an amide group, an imide group, a thioamide group, a sulfonamide group, an enamine group, a triazole, an isoxazole, and the like.
When the position of L is degradable, the drug molecule can realize the polyethylene glycol removal, and the package of polyethylene glycol is released, so that the drug effect can be exerted to the maximum extent.
When degradable linking groups, L may contain degradable linking groups including, but not limited to, any of the above degradable linking groups, specifically including, but not limited to, disulfide bonds, vinyl ether bonds, ester groups, thioester groups, dithioester groups, carbonate groups, thiocarbonate groups, dithiocarbonate groups, trithiocarbonate groups, carbamate groups, thiocarbamate groups, dithiocarbamate groups, acetal groups, cyclic acetals, mercaptals, azaacetals, azacyclanes, dithioketals, dithioacetals, hemiacetals, thiohemiacetals, azahemiacetals, ketals, thioketals, azaketals, thioketals, imine bonds, hydrazone bonds, acylhydrazone bonds, oxime bonds, sulfoximine groups, semicarbazone bonds, thiosemicarbazone bonds, hydrazine groups, hydrazide groups, thiocarbohydrazide groups, azocarbonylhydrazido groups, thiohydrazide groups, A linking group of a thioazo carbonylhydrazide group, a hydrazinoformate group, a hydrazinothiocarbamate group, a carbazide, a thiocarbazohydrazide, an azo group, an isoureido group, an isothioureido group, an allophanate group, a thioallophanate group, a guanidino group, an amidino group, an aminoguanidino group, an imido group, a thioester group, a sulfonate group, a sulfinate group, a sulfonamide group, a sulfonylhydrazide group, a sulfonylurea group, a maleimide group, an orthoester group, a phosphate group, a phosphite group, a hypophosphite group, a phosphonate group, a phosphosilane group, a carbonamide, a thioamide, a phosphoramide, a phosphoramidite, a pyrophosphoryl, a cyclophosphamide, an isocyclophosphamide, a thiophosphoramide, an aconityl group, a peptide bond, a thioamide bond or the like.
L preferably contains any linking group such as triazole, 4, 5-dihydroisoxazole, ether bond, thioether group, amide bond, imide group, imide bond, secondary amine bond, tertiary amine bond, urea bond, ester group, thioester group, disulfide group, thioester group, dithioester group, thiocarbonate group, sulfonate group, sulfonamide group, carbamate group, thiocarbamate group, dithiocarbamate group, hemithioacetal, and carbonate group.
In addition to the degradable or non-degradable linking moieties described above, L may also contain any of the above stably present divalent linking groups STAG, or any combination of two or more of the above stably present divalent linking groups. For example, when modifying the hydroxyl group of a drug molecule, the drug may be modified to attach an amino acid molecule (glycine is the most common, and may be diglycine or polyglycine) to convert the hydroxyl group to an amino group, and the range of functional groups to be reacted with the amino group is wider.
3.3. Reaction between nitrogen-branched nine-arm polyethylene glycol derivative and biologically-relevant substance
Reactions between nitrogen-branched nine-arm polyethylene glycol derivatives and biologically relevant substances include, but are not limited to, those described and exemplified in documents CN104877127A, CN104530413A, CN104530415A, CN 104530417A. For example, CN104530413A corresponds to segments [0936] to [0939 ].
The reaction type between the nitrogen-branched nine-arm polyethylene glycol derivative and the biologically-relevant substance is not particularly limited, and may be a site-specific modification or an unspecified modification (also referred to as a random modification). By way of example, site-directed modifications such as commercial products
Figure BDA0002963906360000761
The site-directed reaction between the N-amino group and the aldehyde group of methionine, such as the site-directed reaction between thiol group and maleimide, vinyl sulfone, 2-iodoacetamide, o-pyridine disulfide, etc., and such as the site-directed reaction between amino group and cyano group and isocyanate, isothiocyanate, etc. By way of example, adventitious modifications such as reactions between amino groups and active esters, commercial products such as
Figure BDA0002963906360000762
Figure BDA0002963906360000763
And (3) performing indefinite-point modification during preparation. The fixed-point modification method and the variable-point modification method described in the literature { Pharm Sci Technol Today,1998,1(8): 352-.
When the nitrogen-branched nine-arm polyethylene glycol derivative modifies the biological related substance, one biological related substance can be connected with 1 or more than 1 nitrogen-branched nine-arm polyethylene glycol derivative molecules. For reference, e.g. commercial products
Figure BDA0002963906360000764
Figure BDA0002963906360000765
One molecule of polyethylene glycol reacts with only one reaction site in one drug molecule; to commercialize the product
Figure BDA0002963906360000766
In this case, one drug molecule may be linked to a plurality of polyethylene glycol molecules. In the present invention, it is preferred that a biologically relevant substance is bound to only one nitrogen-branched nine-arm polyethylene glycol derivative molecule.
When the nitrogen-branched nine-arm polyethylene glycol derivative modifies the bio-related substance with two or more than two reaction sites, under the condition of no specific description, the bio-related substance can react with any one or more reaction sites of the bio-related substance in the same molecule of the bio-related substance modified by the nitrogen-branched nine-arm polyethylene glycol derivative; preferably, 1 molecule of the biologically relevant substance reacts with only 1 functional group.
3.4. Nitrogen-branched nine-arm polyethylene glycol derivative modified small-molecule drug
The invention also discloses a nitrogen-branched nine-arm polyethylene glycol modified micromolecular drug, and D in the corresponding general formula (2) is a residue (SD) of the micromolecular drug. The corresponding preferable structure comprises the general formula (3) and (4) that D is the residue (SD) of the small molecule drug.
SD in the same molecule is derived from the same small molecule drug and can be residues formed after different reaction sites participate in the reaction.
The small molecule drug is a biological related substance with the molecular weight not more than 1000Da, or a small molecule mimicry or an active fragment of any biological related substance.
The small molecule drug may also be a derivative of any one, or a pharmaceutically acceptable salt of any one. The derivatives include, but are not limited to glycosides, nucleosides, amino acids, polypeptide derivatives, in addition to the molecularly modified derivatives.
The type of the small molecule drug is not particularly limited, and can be organic, inorganic, organometallic compounds, oligopeptides or polypeptides and other biologically relevant substances with molecular weight not exceeding 1000 Da. Specifically, the drug composition comprises the small molecule drugs in the class (2), and also comprises biologically-related substances with the molecular weight not exceeding 1000Da in any one of the class (1) and the class (3) to the class (14), and small molecule mimics or active fragments (including variants) of any biologically-related substances.
The molecular weight of the small molecule drug is usually not more than 1000 Da. Any molecular weight in any one range of 0-300 Da, 300-350 Da, 350-400 Da, 400-450 Da, 450-500 Da, 500-550 Da, 550-600 Da, 600-650 Da, 650-700 Da, 700-750 Da, 750-800 Da, 800-850 Da, 850-900 Da, 900-950 Da and 950-1000 Da can be selected; the small value endpoints are not included but the large value endpoints are included in each interval.
The mode of obtaining the small molecule drug is not particularly limited, and includes, but is not limited to, natural extracts and derivatives thereof, degradation products of natural extracts, gene recombination products (molecular cloning products), chemically synthesized substances, and the like.
The hydrophilicity and hydrophobicity of the small molecule drug is not particularly limited, and the small molecule drug may be hydrophilic or water-soluble, or may be hydrophobic or fat-soluble. The charge properties of the small molecule drug are not particularly limited.
The small molecule drug can be the small molecule drug itself, and can also be a dimer or a polymer, a partial subunit or a fragment thereof and the like.
The small molecule drug can be the small molecule drug itself, or a precursor, an activated state, a derivative, an isomer, a mutant, an analogue, a mimetic, a polymorph, a pharmaceutically acceptable salt, a fusion protein, a chemically modified substance, a gene recombinant substance, and the like thereof, and can also be a corresponding agonist, an activator, an inhibitor, an antagonist, a modulator, a receptor, a ligand or ligand, an antibody and a fragment thereof, and the like. The small molecule drug also allows for a target molecule, adjunct or delivery vehicle to be bound to it, either before or after it is bound to the functionalized polyethylene glycol.
The field of application of the small molecule drug is not particularly limited, including but not limited to any of the above-mentioned biologically relevant substances, including but not limited to anticancer drugs, antineoplastic drugs, anti-hepatitis drugs, diabetes treatment drugs, anti-infective drugs, antibiotics, antiviral agents, antifungal drugs, vaccines, anti-respiratory drugs, anti-spasmodics, muscle relaxants, anti-inflammatory drugs, appetite suppressants, migraine treating agents, muscle contractants, antirheumatics, antimalarials, antiemetics, bronchodilators, antithrombotic drugs, antihypertensive drugs, cardiovascular drugs, antiarrhythmic drugs, antioxidants, anti-asthmatic drugs, diuretics, lipid regulators, antiandrogenic drugs, antiparasitic drugs, anticoagulant agents, antineoplastic drugs, hypoglycemic drugs, nutritional agents and additives, growth supplements, anti-inflammatory bowel disease drugs, anti-inflammatory drugs, anti-spasmodics, anti-inflammatory drugs, anti-asthmatics, anti-diabetic drugs, anti-thrombotic drugs, anti-spasmodics, anti-spas, Antibodies, diagnostic agents, contrast agents, and the like. Preferably anticancer, antitumor antibiotic, antiviral or antifungal medicine. Typical anti-cancer or anti-tumor drugs are in accordance with the above.
The small molecular drug is preferably selected from any one or any derivative of SN38, irinotecan, resveratrol, cantharidin and derivatives thereof, chrysin, erycibe extract, flavone or flavonoid drug, red sage root extract and silybum marianum extract or any pharmaceutically acceptable salt; the pharmaceutically acceptable salt can be inorganic salt such as hydrochloride, or organic salt such as oxalate, malate, citrate, etc., preferably hydrochloride. The derivatives include, but are not limited to, glycosides, nucleosides, amino acids, and polypeptide derivatives, in addition to the molecular-modified derivatives. Branching nine-arm polyethylene glycol derivative when nitrogen is availableWhen the perhydroxyl group or the phenolic hydroxyl group is bound to a small molecule drug, an amino acid derivative of the small molecule drug or an oligo-polyethylene glycol fragment having 2 to 10 EO units is preferred, an amino acid derivative of the small molecule drug is more preferred, a glycine or alanine modified product of the small molecule drug is more preferred, a glycine modified product of the small molecule drug is most preferred, that is, L preferably contains an amino acid derivative skeleton, a glycine skeleton or alanine skeleton is more preferred, and a glycine skeleton (-C (═ O) -CH) -is most preferred 2 -NH-), when the reactive group in the amino acid derivative of the small molecule drug is converted to the amino group in the corresponding amino acid. The small molecule drug residue SD includes but is not limited to the fragment [1078 ] of CN104530413A]～[1113]Small molecule drug residues of the fragment.
The nitrogen-branched nine-arm polyethylene glycol derivative and the preparation method thereof according to the present invention will be further described with reference to some specific examples. The specific examples are intended to illustrate the present invention in further detail, and are not intended to limit the scope of the present invention. Among them, in examples of preparing nitrogen-branched nine-arm polyethylene glycol and nitrogen-branched nine-arm polyethylene glycol derivatives, the monodisperse raw materials, key intermediates and products were subjected to molecular weight confirmation by MALDI-TOF. For the determination of the attribution of the characteristic peaks in the nuclear magnetic tests, the analytical methods of CN104877127A, CN104530413A, CN104530415A, CN104530417A and the examples in each cited document were used. The yield in the nitrogen-branched nine-arm polyethylene glycol derivative-modified biologically relevant substance refers to the percentage of the actual mass of the product obtained relative to the theoretical mass.
Example 1: preparation of nitrogen branched nonahydroxyl micromolecule S1-3
Figure BDA0002963906360000771
The preparation process is as follows:
adding 400mL of tetrahydrofuran, a compound S1-1(1.49g,10.0mmol) and excessive diphenyl methyl potassium (75.0mmol) into an anhydrous and oxygen-free closed reaction kettle, then adding a compound S1-2(38.00g,75.0mmol), and reacting at 30 ℃ for 12 hours; completion of the reaction And then, opening the reaction kettle, washing and concentrating the reaction solution, dissolving the reaction solution in methanol, adding 1M hydrochloric acid until the pH value is 3.5, reacting for 4 hours, concentrating, washing and purifying by column chromatography to obtain the nonahydroxyl micromolecule initiator S1-3. The hydrogen spectrum data of the nine-hydroxyl small molecule S1-3 are as follows: 1 H NMR(CDCl 3 )δ(ppm):2.60-2.70(>NCH 2 CH 2 O-, 6H),3.37(-OCH 2 C-,6H),3.48-3.68(>NCH 2 CH 2 O-,6H；-C(CH 2 OH) 3 ,18H)。
Figure BDA0002963906360000781
example 2: preparation of nitrogen-branched nonahydroxyl micromolecule S2-2
Figure BDA0002963906360000782
The preparation process is as follows:
adding 400mL of tetrahydrofuran, a compound S1-1(1.49g,10.0mmol) and excessive diphenyl methyl potassium (75.0mmol) into an anhydrous and oxygen-free closed reaction kettle, then adding a compound S2-1(47.91g,75.0mmol), and reacting at 30 ℃ for 12 hours; after the reaction is finished, the reaction kettle is opened, the reaction solution is washed and concentrated and then dissolved by methanol, 1M hydrochloric acid is added until the pH value is 3.5, and after the reaction is carried out for 4 hours, the nitrogen branched nonahydroxyl micromolecule initiator S2-2 is obtained through concentration, washing and column chromatography purification. The hydrogen spectrum data of the nine-hydroxyl small molecule S2-2 are as follows: 1 H NMR(CDCl 3 )δ(ppm):2.60-2.70 (>NCH 2 CH 2 O-,6H),3.37(-C(CH 2 O-) 4 ,24H),3.47-3.69(>NCH 2 CH 2 O-,6H；-C(CH 2 CH 2 OH) 3 , 36H)。
Figure BDA0002963906360000783
example 3: preparation of nitrogen branched nonahydroxyl micromolecule S3-2
Figure BDA0002963906360000791
The preparation process is as follows:
adding 400mL of tetrahydrofuran, a compound S1-1(1.49g,10.0mmol) and excessive diphenyl methyl potassium (75.0mmol) into an anhydrous and oxygen-free closed reaction kettle, and then adding a compound S3-1(41.30g,75.0mmol), wherein the reaction temperature is 30 ℃, and the reaction time is 12 hours; after the reaction is finished, the reaction kettle is opened, the reaction solution is washed and concentrated and then dissolved by methanol, 1M hydrochloric acid is added until the pH value is 3.5, and after the reaction is carried out for 4 hours, the nitrogen branched nonahydroxyl micromolecule initiator S3-2 is obtained through concentration, washing and column chromatography purification. The hydrogen spectrum data of the nine-hydroxyl small molecule S3-2 are as follows: 1 H NMR(CDCl 3 )δ(ppm):2.60-2.70 (>NCH 2 CH 2 O-,6H),3.37(-OCH 2 C-,6H),3.40-3.78(-OCH 2 CH 2 O-,12H；>NCH 2 CH 2 O-,6H； -C(CH 2 OH) 3 ,18H)。
Figure BDA0002963906360000792
Example 4: preparation of nitrogen branched nonamino micromolecule S4-2
Figure BDA0002963906360000793
The preparation process is as follows:
adding 500mL of tetrahydrofuran, a compound S1-1(1.49g,10.0mmol) and excessive diphenyl methyl potassium (75.0mmol) into a closed anhydrous and oxygen-free reaction kettle, and then adding a compound S4-1(71.56g,75.0mmol), wherein the reaction temperature is 30 ℃, and the reaction time is 12 hours; opening the reaction kettle, washing, concentrating, treating with 20% piperidine/DMF solution, removing Fmoc protection to obtain naked amino group, removing solvent by rotary evaporation, dissolving with dichloromethane, precipitating with anhydrous ether, and precipitating with isopropanolAnd recrystallizing to obtain the nitrogen branched nonamino micromolecule S4-2. The hydrogen spectrum data of the nonamino small molecule S4-2 are as follows: 1 H NMR(CDCl 3 )δ(ppm): 2.60-2.70(>NCH 2 CH 2 O-,6H；-C(CH 2 NH 2 ) 3 ,18H),3.37(-OCH 2 C-,6H),3.48-3.68 (>NCH 2 CH 2 O-,6H)。
Figure BDA0002963906360000794
example 5: preparation of nitrogen-branched nona-armed polyethylene glycol butyne derivative E1-1
Figure BDA0002963906360000801
Corresponding to the general formula (1), wherein U 0 Is composed of
Figure BDA0002963906360000802
L 2 ＝-CH 2 -，F G Is composed of
Figure BDA0002963906360000803
The designed total molecular weight is about 10224Da, wherein each PEG chain has a molecular weight of about 1000Da, corresponding to n 1 ≈n 2 ≈n 3 ≈n 4 ≈n 5 ≈n 6 ≈n 7 ≈n 8 ≈n 9 ≈23。
The preparation process is as follows:
step a: tetrahydrofuran (500mL), a nine-hydroxy small-molecule initiator S1-3(0.93g,2.5mmol) and potassium diphenylmethyl (7.2mmol) were added to a closed anhydrous and oxygen-free reaction vessel in this order.
Step b: adding calculated amount of ethylene oxide, gradually raising the temperature to 60 ℃, and reacting for 48 hours.
Step c: an excess of proton source methanol was added to give a nitrogen branched nine-arm polyethylene glycol H1-1. The hydrogen spectrum data of H1-1 is as follows: 1 H NMR(CDCl 3 )δ(ppm):3.37(-C(CH 2 O-) 4 ,24H),3.55-3.75(-OCH 2 CH 2 OH,36H；-OCH 2 CH 2 O-)。
step d: into a dry, clean 1L round-bottomed flask, N-branched nonamer polyethylene glycol H1-1(19.01g,2.0mmol), excess 4-pentynoic acid (5.30g,54.0mmol) and the solvent dichloromethane (200mL) were added, DMAP (0.06g, 0.5mmol) was added under ice bath, DCC (11.10g,54.0mmol) dissolved in 120mL dichloromethane was added dropwise to the reaction solution, and after completion of dropwise addition, the reaction was carried out at room temperature for 16 hours. After completion of the reaction, insoluble matter was removed by filtration, concentrated and purified by column chromatography to give a nitrogen-branched nonaminorbyl butyne polyethylene glycol derivative E1-1(15.54g, yield 76%). The hydrogen spectrum data of E1-1 is as follows: 1 H NMR(CDCl 3 )δ(ppm): 2.00(-CH 2 CH 2 C≡CH,9H),2.42-2.62(-OC(O)CH 2 CH 2 C≡CH,36H),3.37(-C(CH 2 O-) 4 ,24H), 3.55-3.75(-OCH 2 CH 2 OC(O)-,18H；-OCH 2 CH 2 O-),3.90-4.20(-OCH 2 CH 2 OC(O)-,18H)。
GPC measurement of N-branched nona-armed polyethylene glycol butyne derivative E1-1 to determine M n ≈10.2kDa,PDI＝1.02。
Figure BDA0002963906360000804
Example 6: preparation of nitrogen-branched nine-arm polyethylene glycol azide derivative E2-1
Figure BDA0002963906360000811
Corresponding to the general formula (1), wherein U 0 Is composed of
Figure BDA0002963906360000812
L 2 ＝-OCH 2 CH 2 -，F G Is composed of
Figure BDA0002963906360000813
Designed overall molecular weightAbout 19738Da, wherein each PEG chain has a molecular weight of about 2000Da corresponding to n 1 ≈n 2 ≈n 3 ≈n 4 ≈n 5 ≈n 6 ≈n 7 ≈n 8 ≈n 9 ＝n+1＝46。
The preparation process is as follows:
dissolving the nonahydroxyl micromolecule S2-2(0.9g,1.0mmol) in DMF in an anhydrous and oxygen-free closed reaction kettle, and then adding potassium carbonate (0.68g,4.95mmol) and benzenesulfonyl polyethylene glycol azide derivative dissolved in DMF
Figure BDA0002963906360000814
(S6-1,30.58g, 13.5mmol), the reaction was stirred at 80 ℃ for 24 hours. After the reaction is finished, the reaction liquid is cooled to room temperature, then insoluble substances are removed by filtration, the mixture is concentrated and extracted, the organic phase is washed by brine, dried by anhydrous sodium sulfate, filtered, concentrated and purified by silica gel column chromatography, and the nitrogen-branched nine-arm polyethylene glycol azide derivative E2-1(10.86g) is obtained. The hydrogen spectrum data of E2-1 is as follows: 1 H NMR(CDCl 3 )δ(ppm):3.30-3.40(-OCH 2 CH 2 N 3 ,18H；C(CH 2 O-) 4 , 24H),3.45-3.80(-OCH 2 CH 2 N 3 ,18H；-OCH 2 CH 2 O-)。
GPC measurement of N-branched nine-arm polyethylene glycol azide derivative E2-1 to determine M n ≈19.7kDa,PDI＝1.02。
Figure BDA0002963906360000815
Example 7: preparation of nitrogen-branched nona-armed polyethylene glycol ethylamine derivative E3-1
Figure BDA0002963906360000821
Corresponding to the general formula (1), wherein U 0 Is composed of
Figure BDA0002963906360000822
L 2 ＝-OC(O)OCH 2 CH 2 -，F G Is composed of
Figure BDA0002963906360000823
The designed total molecular weight is about 28609Da, wherein each PEG chain has a molecular weight of about 3000Da, corresponding to n 1 ≈n 2 ≈n 3 ≈n 4 ≈n 5 ≈n 6 ≈n 7 ≈n 8 ≈n 9 ≈n≈68。
The preparation process is as follows:
step a: in a dry clean 1000mL round bottom flask, S7-1(1.91g, 10.0mmol) was dissolved in dichloromethane, N-hydroxysuccinimide (NHS,4.11g,36.0mmol) was added and dissolved with stirring, dicyclohexylcarbodiimide (DCC,11.10g,54.0mmol) was added to the solution and reacted overnight under nitrogen. Then, S7-2(11.14g, 33.0 mmol) was added to the reaction solution, and the reaction was stirred for 5 hours. After the reaction was completed, insoluble matter was removed by filtration, washed, concentrated and dissolved in methanol, 1M hydrochloric acid was added to adjust the pH to 3.5, and after 4 hours of reaction, the mixture was concentrated, washed and subjected to column chromatography, thereby obtaining nitrogen-branched nonahydroxy small molecule S7-3 (4.05 g). The hydrogen spectrum data of the nine-hydroxyl small molecule S7-3 are as follows: 1 H NMR(DMSO-d 6 )δ(ppm):3.26 (N(CH 2 CONH-) 3 ,18H),3.72(-C(CH 2 OH) 3 ,18H)。
Step b: in a dry clean 1000mL round bottom flask, amino Fmoc protected polyethylene glycol succinimide carbonate
Figure BDA0002963906360000824
(S7-4, azeotropic removal of water with toluene, 3460Da, PDI 1.02,10.8 mmol) was dissolved in dichloromethane, 4-Dimethylaminopyridine (DMAP) (0.13g,1.1mmol) was added thereto, and the mixture was stirred and mixed, and nonahydroxy small molecule S7-3(0.50g,1.0mmol) was added to the reaction mixture, followed by stirring at room temperature for 16 hours. After the reaction was completed, the reaction solution was spin-dried, recrystallized from isopropanol, and purified by ion exchange resin to obtain amino Fmoc-protected N-branched nine-arm polyethylene glycol amino derivative S7-5(21.43g, yield 70%). The hydrogen spectrum data of S7-5 is as follows: 1 H NMR(CDCl 3 )δ(ppm):3.00-3.20 (-OCH 2 CH 2 NH-,6H),3.45-3.75(-OCH 2 CH 2 NH-,18H；-OCH 2 CH 2 O-；-C(O)OCH 2 CH 2 O-,18H), 3.90-4.20(-CH 2 OC(O)OCH 2 -,36H)。
step c: deprotecting S7-5 containing Fmoc protected amino, treating S7-5 with 20% piperidine/DMF solution, removing the solvent by rotary evaporation, dissolving with dichloromethane, precipitating with anhydrous ether, and recrystallizing with isopropanol to obtain the N-branched nona-armed polyethylene glycol ethylamine derivative E3-1 containing nine naked amino groups. And hydrogen spectrum nuclear magnetic testing shows that the characteristic peaks of the Fmoc aromatic ring, 9-H and methylene disappear, and the terminal group substitution rate is about 100%. The hydrogen spectrum data of E3-1 is as follows: 1 H NMR(CDCl 3 )δ(ppm):2.82-2.95(-OCH 2 CH 2 NH 2 , 18H)。
GPC measurement of N-branched nona-PEGylethylamine derivative E3-1 to determine M n ≈28.6kDa,PDI＝1.03。
Figure RE-GDA0003038345320000831
Example 8: preparation of nitrogen-branched nine-arm polyethylene glycol maleimide derivative E4-2
Figure BDA0002963906360000832
Corresponds to the general formula (1), wherein U 0 Is composed of
Figure BDA0002963906360000833
L 2 ＝-CH 2 CH 2 -，F G Is composed of
Figure BDA0002963906360000834
The designed total molecular weight is about 38417Da, wherein each PEG chain has a molecular weight of about 4000Da corresponding to n 1 ≈n 2 ≈n 3 ≈n 4 ≈n 5 ≈n 6 ≈n 7 ≈n 8 ≈n 9 ≈n≈91。
The preparation process is as follows:
step a: s1-1(1.49g,10.0mmol), an excess of S8-1(23.35g, 45.0mmol) and the solvent dichloromethane (200mL) were charged into a dry, clean 1-L round-bottomed flask, DMAP (0.05g,0.45mmol) was added, DCC (9.25g,45.0mmol) dissolved in 100mL of dichloromethane was added dropwise to the reaction solution, and after completion of the addition, the reaction was carried out at room temperature for 16 hours. After the reaction, insoluble substances are removed by filtration, the mixture is concentrated, tetrahydrofuran is dissolved, tetra-tert-butylammonium fluoride (TBAF) is added, the reaction is carried out overnight, TBS protection is removed, and silica gel column chromatography purification is carried out to obtain the nitrogen branched nonahydroxy micromolecule S8-2(4.55 g). The molecular weight of the nonahydroxyl small molecule S8-2 is determined to be 623Da by MALDI-TOF test.
Step b: in a water-free and oxygen-free closed reaction vessel, S8-2(0.62g,1.0mmol) was dissolved in DMF, followed by addition of potassium carbonate (4.15g,30.0mmol) and benzenesulfonylated polyethylene glycol furan protected maleimide derivative dissolved in DMF
Figure BDA0002963906360000841
(S8-3,59.93g,13.5mmol), the reaction was stirred at 80 ℃ for 24 hours. After the reaction, the reaction solution was cooled to room temperature, and then insoluble matter was removed by filtration, concentrated, extracted, and the combined organic phase was washed with brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to obtain nitrogen-branched nona-armed polyethylene glycol furan-protected maleimide derivative E4-1(22.25 g). The hydrogen spectrum data of E4-1 is as follows: 1 H NMR(CDCl 3 )δ(ppm):2.61-2.68 (-C(O)CH 2 CH 2 N<,18H),2.85(-COCHCHCO-,18H),3.42-3.76(-OCH 2 CH 2 OC(O)-,18H； -OCH 2 CH 2 O-),3.78-3.85(-C(O)CH 2 CH 2 N<,18H),3.90-4.20(-OCH 2 CH 2 OC(O)-,18H),5.22 (-CHOCH-,18H),6.49(-CH＝CH-,18H)。
Step c: and (4) removing the furan protecting group of the maleimide group. In a dry clean 1000mL round bottom flask, E4-1 was dissolved in 400mL toluene and 2, 6-di-tert-butyl-4-methylphenol (BHT, antioxidant) was added4g) heating to 125 ℃, stirring and reacting for 5h, concentrating and recrystallizing with isopropanol to obtain the nitrogen-branched nona-armed polyethylene glycol maleimide derivative with the structure shown as E4-2. Hydrogen spectrum test showed that peaks (2.85 (-COCHCO, 18H),5.22(-CHOCH, 18H),6.49 (-CH ═ CH-,18H)) on the E4-2 furan ring disappeared and peaks on the maleimide ring appeared, and the data of hydrogen spectrum were as 1 H NMR(CDCl 3 )δ(ppm)： 6.70(-CH＝CH-,18H)。
GPC measurement of N-branched nine-arm polyethylene glycol maleimide derivative E4-2 to determine M n ≈38.4kDa,PDI ＝1.03。
Figure RE-GDA0003038345320000842
Example 9: preparation of nitrogen-branched nine-arm polyethylene glycol hydroxyl derivative E5-2
Figure BDA0002963906360000851
Corresponding to the general formula (1), wherein U 0 Is composed of
Figure BDA0002963906360000852
L 2 ＝-OC(O)NHCH 2 CH 2 -，F G Is composed of
Figure BDA0002963906360000853
The designed total molecular weight is about 46531Da, wherein each PEG chain has a molecular weight of about 5000Da, corresponding to n 1 ≈n 2 ≈n 3 ≈n 4 ≈n 5 ≈n 6 ≈n 7 ≈n 8 ≈n 9 ≈n≈113。
The preparation process is as follows:
step a in a dry clean 1000mL round-bottom flask, S9-1(1.46g, 10.0mmol) was dissolved in dichloromethane, N-hydroxysuccinimide (NHS,4.11g,36.0mmol) was added and stirred to dissolve, dicyclohexylcarbodiimide (DCC,11.10g,54.0mmol) was added to the solution and the reaction was carried out under nitrogen protection Should be allowed to stand overnight. Then, S9-2(12.75g, 33.0 mmol) was added to the reaction solution, and the reaction was stirred for 5 hours. After the reaction was completed, insoluble matter was removed by filtration, washed, concentrated and dissolved in methanol, 1M hydrochloric acid was added to adjust the pH to 3.5, and after 4 hours of reaction, the mixture was concentrated, washed and subjected to column chromatography, thereby obtaining nitrogen-branched nonahydroxy small molecule S9-3 (4.40 g). The hydrogen spectrum data of the nine-hydroxyl small molecule S9-3 are as follows: 1 H NMR(CDCl 3 )δ(ppm):2.62-2.68 (N(CH 2 CH 2 -) 3 ,6H),3.25-3.32(N(CH 2 CH 2 NH-) 3 ,6H),6.76-6.83(Ar-H,6H)。
step b: in a dry clean round-bottom flask, S9-3(0.30g,0.5mmol) containing nine naked hydroxyl groups was added, dissolved in dichloromethane, and under nitrogen protection, the catalyst dibutyltin dilaurate (1.0mL) was added, and then under ice bath, the polyethylene glycol isocyanate derivative containing TBS protected hydroxyl groups was slowly added dropwise
Figure BDA0002963906360000854
(S9-4, 5kDa, PDI 1.02,10.8mmol) was azeotropically removed with toluene, and after completion of the dropwise addition, the reaction was stirred at room temperature for 8 hours. After the reaction was complete, excess activated silica gel was added, filtered, concentrated, and column purified to afford the nitrogen-branched nine-armed polyethylene glycol derivative E5-1(15.46g, 65% yield) containing nine TBS protected hydroxyl groups. The hydrogen spectrum data of E5-1 is as follows: 1 H NMR(CDCl 3 )δ(ppm):0.21(-Si(CH 3 ) 2 -, 54H),0.98(-SiC(CH 3 ) 3 ,81H),3.00-3.20(-NHCH 2 CH 2 O-,18H),3.42-3.76(-OCH 2 CH 2 O-； -NHCH 2 CH 2 O-,18H)。
step c: and (3) removing the TBS protecting group, dissolving E5-1 in tetrahydrofuran in a dry and clean round-bottom flask, adding tetra-tert-butylammonium fluoride (TBAF), reacting overnight, removing the TBS protection, and purifying by silica gel column chromatography to obtain the nitrogen-branched nine-arm polyethylene glycol hydroxyl derivative E5-2. Nuclear magnetic test TBS characteristic peaks disappeared.
GPC measurement of N-branched nine-arm hydroxyl derivative E5-2 of polyethylene glycol to determine M n ≈46.5kDa,PDI＝1.03。
Figure RE-GDA0003038345320000861
Example 10: preparation of nitrogen-branched nine-arm polyethylene glycol hydroxyl derivative with branched structure at branch chain end
Figure BDA0002963906360000862
Corresponding to the general formula (1), wherein U 0 Is composed of
Figure BDA0002963906360000863
L 2 ＝-C(O)OCH 2 CH 2 -，F G Is composed of
Figure BDA0002963906360000864
The designed total molecular weight is about 56450Da, wherein each PEG chain has a molecular weight of about 6000Da corresponding to n 1 ≈n 2 ≈n 3 ≈n 4 ≈n 5 ≈n 6 ≈n 7 ≈n 8 ≈n 9 ≈n≈136。
Step a: in a dry clean 1000mL round bottom flask, hydroxy TBS protected polyethylene glycol succinimide carbonate
Figure BDA0002963906360000871
(S10-1, 6.3kDa, PDI 1.02 and 10.8mmol) was dissolved in dichloromethane, 4-dimethylaminopyridine DMAP (0.13g and 1.1mmol) was added thereto, the mixture was stirred and mixed, nonamino small molecule S4-2(0.49g and 1.0mmol) was added to the reaction mixture, and the mixture was stirred at room temperature for 16 hours. After the reaction, the reaction solution was spin-dried, recrystallized from isopropanol, and purified by ion exchange resin to obtain hydroxy TBS-protected nitrogen-branched nona-armed polyethylene glycol hydroxy derivative S10-2(40.44g, 72% yield). The hydrogen spectrum data of S10-2 is as follows: 1 H NMR(CDCl 3 )δ(ppm):0.21(-Si(CH 3 ) 2 -, 54H),0.98(-SiC(CH 3 ) 3 ,81H),(-C(CH 2 NHC(O)O-) 3 ,18H),3.45-3.75(-NHC(O)OCH 2 CH 2 O-, 18H；-OCH 2 CH 2 O-),4.14-4.38(-NHC(O)OCH 2 CH 2 O-,18H)。
step b: and (3) removing the TBS protecting group, dissolving S10-2 in tetrahydrofuran in a dry and clean round-bottom flask, adding tetra-tert-butylammonium fluoride (TBAF), reacting overnight, removing the TBS protection, and purifying by silica gel column chromatography to obtain the N-branched nine-arm polyethylene glycol hydroxyl derivative S10-3. S10-3 showed disappearance of characteristic peaks for TBS by nuclear magnetic testing.
Step c: a dry clean 1L round bottom flask was charged with nonamer polyethylene glycol hydroxy derivative S10-3(27.57g,0.5 mmol), excess S10-4(3.53g,9.0mmol) and solvent dichloromethane (300mL), DMAP (0.01 g,0.12mmol) was added under ice bath conditions, DCC (1.85g,9.0mmol) dissolved in 100mL dichloromethane was added dropwise to the reaction mixture, and after completion of addition, the reaction was carried out at room temperature for 16 h. After the reaction was completed, insoluble matter was removed by filtration, concentrated, and purified by column chromatography to obtain hydroxy TBS-protected nitrogen-branched nine-arm polyethylene glycol intermediate S10-5(21.94g, yield 75%). The hydrogen spectrum data of S10-5 is as follows: 1 H NMR (CDCl 3 )δ(ppm):0.21(-Si(CH 3 ) 2 ,108H),0.98(-SiC(CH 3 ) 3 ,162H),3.40-3.46(>NCH 2 C(O)O-, 18H)。
step d: and (3) removing the TBS protecting group, dissolving S10-5 in tetrahydrofuran in a dry and clean round-bottom flask, adding tetra-tert-butylammonium fluoride (TBAF), reacting overnight, removing the TBS protection, and purifying by silica gel column chromatography to obtain the nitrogen-branched nine-arm polyethylene glycol hydroxyl derivative E6-1 with the branched chain end as a branched structure. The E6-1 nuclear magnetic test showed the disappearance of the characteristic peaks of TBS, and the hydrogen spectrum data of E6-1 were as follows: 1 H NMR(CDCl 3 )δ(ppm):3.76-3.80(-N(CH 2 CH 2 OH) 2 ,36H)。
GPC measurement was carried out on nitrogen-branched nine-arm polyethylene glycol hydroxy derivative E6-1 having a branched structure at the end of the branch, and M was determined n ≈56.5kDa,PDI＝1.03。
Figure RE-GDA0003038345320000872
Example 11: preparation of nitrogen-branched nine-arm polyethylene glycol derivative E7-2 with branched structure at branch chain end
Figure BDA0002963906360000881
Corresponding to the general formula (1), wherein U 0 Is composed of
Figure BDA0002963906360000882
L 2 Is absent, F G Is composed of
Figure BDA0002963906360000883
(g＝1,k＝3,L 0 ＝-COCH 2 CH 2 CONH-,
Figure BDA0002963906360000884
q＝0,Z 2 Is not present, q 1 ＝0,Z 1 Is absent, R 01 ＝OPG 4 ,PG 4 TBS). The designed total molecular weight is about 14417Da, wherein each PEG chain has a molecular weight of about 1000Da corresponding to n 1 ≈n 2 ≈n 3 ≈n 4 ≈n 5 ≈n 6 ≈n 7 ≈n 8 ≈n 9 ≈23。
The preparation process is as follows:
step a: nine-arm polyethylene glycol H1-1(9.50g,1.0mmol) was dissolved in toluene (300mL), and excess succinic anhydride (4.50g,45.0mmol) was added at 50 ℃ for 12 hours. After the reaction was completed, the reaction vessel was opened, the solvent was concentrated, and then precipitated in anhydrous ether at 0 ℃, filtered, dried, and purified with silica gel column to obtain nitrogen-branched nona-armed polyethylene glycol propionic acid derivative E7-1(8.12g, yield 78%). The hydrogen spectrum data of E7-1 is as follows: 1 H NMR(CDCl 3 )δ(ppm):2.58-2.64 (-C(＝O)CH 2 CH 2 COOH,36H),3.45-3.85(-OCH 2 CH 2 OC(O)-,18H；-OCH 2 CH 2 O-),4.10-4.25 (-OCH 2 CH 2 OC(O)-,18H)。
step b: 2.08g of nitrogen-branched nonaarm polyethylene glycol propionic acid derivative E7-1 (azeotropic dehydration of toluene), 20mL of triethylamine and 83.51g of compound S11-1 containing three TBS protected hydroxyl groups and one naked amino group are added into a dry and clean 1-L round-bottom flask, solvent dichloromethane (400mL) is added under the protection of nitrogen, the mixture is stirred until the mixture is dissolved, 44.41g of Dicyclohexylcarbodiimide (DCC) is added, reaction is carried out for 24 hours at room temperature, insoluble substances are removed by filtration, concentration and isopropanol recrystallization are carried out, so as to obtain the nitrogen-branched nonaarm polyethylene glycol derivative E7-2 with a three-branched structure at the tail end of a branched chain. The hydrogen spectrum data of E7-2 is as follows: 1 H NMR(CDCl 3 )δ (ppm):0.21(-Si(CH 3 ) 2 ,162H),0.98(-SiC(CH 3 ) 3 ,243H),2.80-3.00(-C(O)CH 2 CH 2 C(O)-,36H), 3.40-3.80(-OCH 2 CH 2 O-),3.90-4.20(-NHC(CH 2 O-) 3 ,54H)。
GPC measurement of N-branched nine-arm polyethylene glycol derivative E7-2 having three-branched structure at the end of branched chain, M was determined n ≈14.4kDa,PDI＝1.03。
Figure BDA0002963906360000891
Example 12: preparation of nitrogen-branched nine-arm polyethylene glycol derivative E8-1 with comb-shaped branched chain end
Figure BDA0002963906360000892
Corresponding to the general formula (1), wherein U 0 Is composed of
Figure BDA0002963906360000893
L 2 ＝-CH 2 -，F G Is composed of
Figure BDA0002963906360000894
The designed total molecular weight is about 39888Da, wherein each PEG chain has a molecular weight of about 1000Da corresponding to n 1 ≈n 2 ≈n 3 ≈n 4 ≈n 5 ≈n 6 ≈n 7 ≈n 8 ≈n 9 ≈n 10 ≈23。
The preparation process is as follows:
step a: 200mL of tetrahydrofuran, 9-hydroxyl nitrogen-branched nonaarm polyethylene glycol H1-1(19.01g,2.0mmol) and diphenylmethyl potassium (14.4mmol) were added sequentially to a water-free and oxygen-free closed reaction vessel.
Step b: a calculated amount of S12-1(Ethoxy ethyl glycosyl ether,2435mmol) was added and the temperature was gradually raised to 60 ℃ for 48 hours.
Step c: adding excessive diphenyl methyl potassium (36.0mmol), then adding excessive methyl iodide (90.0mmol), and reacting at 30 ℃ for 12 hours; opening the reaction kettle, concentrating the solvent, precipitating in anhydrous ether at 0 ℃, filtering and drying to obtain a nitrogen-branched nine-arm polyethylene glycol derivative E8-1 with a comb-shaped branched chain end; the hydrogen spectrum data of E8-1 is as follows: 1 H NMR(CDCl 3 )δ(ppm):1.22(-OCH 2 CH 3 ),1.36(-OCH(O)CH 3 ),3.35(-OCH 3 ),3.40-3.80 (-OCH 2 CH 2 O-,-OCH(CH 2 O-) 2 ,OCH 2 CH 3 ),4.75(-OCHCH 3 (OCH 2 ))。
GPC test was carried out on nitrogen-branched nine-arm polyethylene glycol derivative E8-1 having branched chain ends of comb-like structure, and M was determined n ≈39.9 kDa,PDI＝1.04。
Figure BDA0002963906360000901
Example 13: preparation of nitrogen-branched nine-arm polyethylene glycol derivative E9-1 with hyperbranched structure at branch chain end
Figure BDA0002963906360000902
Corresponding to the general formula (1), wherein U 0 Is composed of
Figure BDA0002963906360000903
L 2 ＝-CH 2 -，F G See the above structural formula. The designed total molecular weight is about 23972Da, wherein each PEG chain has a molecular weight of about 1000Da corresponding to n 1 ≈n 2 ≈n 3 ≈n 4 ≈n 5 ≈n 6 ≈n 7 ≈n 8 ≈n 9 ≈23。
The preparation process is as follows:
step a: into a water-free and oxygen-free closed reaction vessel were added 200mL of tetrahydrofuran, nitrogen-branched nonaarm polyethylene glycol H1-1 containing 9 hydroxyl groups (19.01g,2.0mmol), and diphenylmethyl potassium (14.4mmol) in this order.
Step b: a calculated amount of compound S13-1(3600mmol) was added, the temperature was gradually raised to 60 ℃ and the reaction was carried out for 48 hours.
Step c: adding excessive methanol, concentrating the solvent, precipitating in 0 deg.C anhydrous ether, filtering, and drying to obtain nitrogen-branched nona-armed polyethylene glycol derivative E9-1 with branched chain end having hyperbranched structure; the hydrogen spectrum data of E9-1 is as follows: 1 H NMR(CDCl 3 ) δ(ppm):3.40-3.80(-OCH 2 CH 2 O-；-OCH(CH 2 O-) 2 ；-OCH 2 CH(O)CH(O)-),3.85-4.40 (-OCH 2 CH(O)CH(O)-)。
GPC measurement is carried out on nitrogen branched nine-arm polyethylene glycol derivative E9-1 with branched chain ends of hyperbranched structure, and M is determined n ≈24.0kDa,PDI＝1.03。
Figure BDA0002963906360000911
Example 14: preparation of nitrogen-branched nine-arm polyethylene glycol derivative E10-1 with hyperbranched structure at branch chain end
Figure BDA0002963906360000912
Corresponding to the general formula (1), wherein U 0 Is composed of
Figure BDA0002963906360000913
L 2 ＝-CH 2 -，F G See the above structural formula. The designed total molecular weight is about 20036Da, wherein, the molecular weight of each PEG chain is about 1000Da, corresponding to n 1 ≈n 2 ≈n 3 ≈n 4 ≈n 5 ≈n 6 ≈n 7 ≈n 8 ≈n 9 ≈23。
The preparation process is as follows:
step a: 200mL of tetrahydrofuran, nitrogen-branched nine-armed polyethylene glycol H1-1(19.01g,2.0 mmol) containing 9 hydroxyl groups, and diphenylmethyl potassium (14.4mmol) were added sequentially.
Step b: a calculated amount of glycidol S14-1(3600mmol) was added and the temperature was gradually raised to 60 ℃ for 48 hours.
Step c: adding excessive methanol, concentrating the solvent, precipitating in 0 deg.C anhydrous ether, filtering, and drying to obtain nitrogen-branched nine-arm polyethylene glycol E10-1 with branched chain end having hyperbranched structure; the hydrogen spectrum data of E10-1 is as follows: 1 H NMR(CDCl 3 )δ (ppm):3.40-3.85(-CH 2 CH 2 O-,-OCH(CH 2 O-) 2 )。
GPC measurement is carried out on nitrogen branched nine-arm polyethylene glycol E10-1 with branched chain ends of hyperbranched structures, and M is determined n ≈20.0 kDa,PDI＝1.03。
Figure BDA0002963906360000921
Example 15: preparation of nitrogen-branched nine-arm polyethylene glycol derivative E11-2 with branch chain end of dendritic structure
Figure BDA0002963906360000922
Corresponding to the general formula (1), wherein U 0 Is composed of
Figure BDA0002963906360000923
L 2 ＝-CH 2 -，F G See the above structural formula. Wherein the designed total molecular weight is about 14171Da, wherein each PEG chain has a molecular weight of about 1000Da corresponding to n 1 ≈n 2 ≈n 3 ≈n 4 ≈n 5 ≈n 6 ≈n 7 ≈n 8 ≈n 9 ≈23。
The preparation process is as follows:
step a: adding 200mL of tetrahydrofuran, nitrogen-branched nine-arm polyethylene glycol H1-1(19.01g,2.0 mmol) and excessive diphenyl methyl potassium (144.0mmol) into an anhydrous and oxygen-free closed reaction kettle, and then adding excessive compound S15-1(360 mmol), wherein the reaction temperature is 30 ℃, and the reaction time is 12 hours; opening the reaction kettle, concentrating the solvent, precipitating in anhydrous ethyl ether at 0 ℃, filtering and drying to obtain a nitrogen-branched nine-arm polyethylene glycol intermediate S15-2 protected by hydroxyl silyl ether at the end part; the hydrogen spectrum data of S15-2 is as follows: 1 H NMR(CDCl 3 )δ(ppm):0.21(-Si(CH 3 ) 2 ),0.98(-SiC(CH 3 ) 3 ),2.90-3.10 (-OCHCH 2 OSi-),3.40-3.80(-CH 2 CH 2 O-,-OCH(CH 2 O-) 2 ),3.80-4.10(-OCHCH 2 OSi-)。
Step b: and (3) removing the TBS protecting group, adding the intermediate S15-2 prepared in the previous step into a dry and clean container, dissolving with tetrahydrofuran, adding tetra-tert-butylammonium fluoride (TBAF), and reacting overnight to obtain the nitrogen-branched nona-armed polyethylene glycol intermediate E11-1 containing 18 exposed hydroxyl groups.
Step c: and (c) repeating the steps a and b twice to obtain the nitrogen branched nine-arm polyethylene glycol derivative E11-2 with the branch chain end of a dendritic structure.
GPC measurement of N-branched nine-arm polyethylene glycol derivative E11-2 having branched chain ends of dendritic structure to determine M n ≈ 14.2kDa,PDI＝1.03。
Figure RE-GDA0003038345320000931
Example 16: preparation of nine-arm polyethylene glycol propionaldehyde derivative E12-1
Figure BDA0002963906360000932
Corresponding to the general formula (1), wherein U 0 Is composed of
Figure BDA0002963906360000933
L 21 ＝-C(O)NHCH 2 CH 2 -，F G Is composed of
Figure BDA0002963906360000934
The designed total molecular weight is about 5920Da, wherein each PEG chain has a molecular weight of about 500Da, corresponding to n 1 ≈n 2 ≈n 3 ≈n 4 ≈n 5 ≈n 6 ≈n 7 ≈n 8 ≈n 9 ＝n＝11。
The preparation process is as follows:
step a: adding 400mL of tetrahydrofuran, a compound S1-1(1.49g,10.0 mmol) and excessive diphenyl methyl potassium (75.0mmol) into an anhydrous and oxygen-free closed reaction kettle, and then adding a compound S16-1(38.60g,75.0mmol) into the reaction kettle, wherein the reaction temperature is 30 ℃, and the reaction time is 12 hours; after the reaction was complete, the reaction vessel was opened, and the reaction was washed, concentrated and stirred with TFA overnight at room temperature. The excess TFA was removed and purified by silica gel column chromatography to give nine functionalized small molecule S16-2 containing nine naked carboxyl groups. The molecular weight of S16-2 was determined to be 671Da by MALDI-TOF test.
Step b: adding polyethylene glycol ethylamine derivative with acetal group at one end into a dry and clean 1L round-bottom flask
Figure BDA0002963906360000941
(S16-3, 500Da, PDI 1.01,21.6mmol) and a nine-functional small molecule intermediate S16-2(0.67g,1.0mmol) under the protection of nitrogen, adding solvent dichloromethane 300mL, stirring to dissolve, then adding triethylamine 10mL and dicyclohexylcarbodiimide 10g in sequence(DCC) and a small amount of 4-Dimethylaminopyridine (DMAP) were reacted at room temperature for 24 hours, and then insoluble matter was removed by filtration, concentrated, recrystallized from isopropanol, and purified by column to obtain a nitrogen-branched nona-armed polyethylene glycol acetal derivative S16-4(5.01g, yield 76%). The hydrogen spectrum data of S16-4 are as follows: 1 H NMR(CDCl 3 )δ(ppm):1.14-1.34(-OCH 2 CH 3 ,54H),2.67(-CCH 2 CONH-,12H),3.43-3.80 (-OCH 2 CH 3 ,36H；-OCH 2 CH 2 O-),4.49-4.69(>CH 2 CH 2 CH<,9H)。
step c: acetal deprotection of S16-4, dissolving S16-4 in dichloromethane in a dry clean 1000mL round bottom flask, and adding dropwise acetic acid solution to the solution to bring the solution to pH 3-4, followed by stirring at room temperature for 21 hours. After the reaction is finished, freeze-drying overnight, and purifying to obtain the nitrogen-branched nine-arm polyethylene glycol propionaldehyde derivative E12-1. The hydrogen spectrum data of E12-1 are as follows: 1 H NMR(CDCl 3 )δ(ppm):9.77(-CH 2 CH 2 CHO,9H)。
GPC measurement of N-branched nine-arm polyethylene glycol propionaldehyde derivative E12-1 to determine M n ≈5.9kDa,PDI＝1.01。
Figure BDA0002963906360000942
Example 17: preparation of nine-arm polyethylene glycol modified irinotecan derivative E13-1
Figure BDA0002963906360000951
Into a dry clean 1L round bottom flask, nitrogen-branched nine-arm polyethylene glycol H1-1(19.01g,2.0mmol), excess irinotecan propionic acid derivative S17-1(18.16g,27.0mmol) and solvent dichloromethane (200mL) were added, DMAP (0.04g,0.4mmol) was added under ice bath conditions, DCC (4.93g,24.0mmol) dissolved in 100mL dichloromethane was added dropwise to the reaction solution, and after the addition, the reaction was carried out at room temperature for 16H. After the reaction was completed, insoluble matter was removed by filtration, concentrated, and purified by column chromatography to obtain nitrogen-branched nine-arm polyethylene glycol-modified irinotecan derivative E13-1(23.09g, yield 75%). The structure of E13-1 was determined by NMR.
GPC measurement of N-branched nine-arm polyethylene glycol irinotecan derivative E13-1 to determine M n ≈10.0kDa,PDI＝ 1.02。
Figure BDA0002963906360000952
Example 18: biological testing of nine-armed pegylated irinotecan
(1) Cytotoxicity assays
The cytotoxicity test of the nine-arm pegylated irinotecan E13-1 is carried out by adopting an MTT staining method, a blank control group and a positive control group are set in the experimental process, the blank control group is only added with a culture medium without any medicament, the positive control group is added with a single irinotecan medicament with a certain concentration, the experimental group is added with the nine-arm pegylated irinotecan medicament with corresponding concentration, the medicament concentration is three gradient concentration points of 1nM, 10 nM and 100nM, and each concentration is 6 multiple holes, and the experiment is repeated for three times. COLO205 human colon cancer cells, human colon adenocarcinoma cells HT29 cells, human lung adenocarcinoma cells A549 cells, pancreatic cancer cells MiaPaCa-2 cells, human ovarian cancer cells A2780 cells and human ovarian adenocarcinoma cells OVCAR-3 cells are selected as in-vitro cancer cell models.
At a seed density of 1X 10 4 Cells per well, 100. mu.L/well of cell suspension was seeded into 96-well plates. After inoculation, 4% CO at 37 ℃ 2 The cell culture box is used for incubation and culture for 24 hours. Dissolving the drugs in a culture medium to prepare the required concentration, adding a proper amount of cosolvent if necessary, removing the old culture medium, adding 100 mu L of culture medium containing drugs with the concentrations of 1nM, 10nM and 100nM into each experimental group, adding 100 mu L of fresh culture medium into a blank control group, adding irinotecan with the corresponding concentration into a positive control group, adding nine-arm pegylated irinotecan with the corresponding concentration into the experimental group, and adding 6 multiple wells into each group. After the drug was incubated with the cancer cells for 48h, 20. mu.L of MTT in PBS buffer at 5mg/mL was added to each well. MTT and cancerAfter the cells were incubated for 4 hours, the mixed solution of the medium and MTT buffer was aspirated, DMSO 150 μ L/well was added to dissolve formazan, a purple crystal of living cells, and the 96-well plate was shaken gently to dissolve formazan sufficiently, so that the solution in the well did not overflow into another well and the assay result was not affected by shaking, and after shaking sufficiently, absorbance at 490nm was measured with a microplate reader. The results of calculation of the plots based on the measured absorbance values show that the irinotecan group and the nine-arm pegylated irinotecan group have significant cancer cell proliferation inhibition effects on the above six cancer cells compared with the blank control group; compared with the positive control group, namely the irinotecan group alone, the nine-arm pegylated irinotecan group also shows stronger cancer cell proliferation inhibition effect on the six cancer cells.
(2) Antitumor effect
Using animal transplantable tumor experimental method with H 22 The mouse liver cancer cells are inoculated to the right axilla of the mouse to form solid tumors, and the administration is carried out by tail vein injection respectively 2 days and 7 days after the inoculation, wherein the administration mode is single administration. After 2 weeks of dosing, mice were sacrificed by cervical dislocation, tumor was detached, and weighed. The results show that compared with a blank control, irinotecan and the nine-arm pegylated irinotecan both have obvious tumor inhibition effects, and the tumor inhibition rate of the nine-arm pegylated irinotecan in the experimental group is remarkably higher than that of irinotecan alone in the positive control group.
The above description is only an embodiment of the present invention, and not intended to limit the scope of the present invention, and all modifications of equivalent structures and equivalent processes, which are made by the present specification, or directly or indirectly applied to other related technical fields, are included in the scope of the present invention.
It will be apparent to those skilled in the art that the invention can be practiced in a wide range of equivalent parameters, concentrations, and conditions without departing from the spirit and scope of the invention and without undue experimentation. While the invention has been described with reference to specific embodiments, it will be understood that it is capable of further modifications. In general, this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure as come within known or customary practice within the art to which the invention pertains.

Claims (31)

1. A nitrogen-branched nine-arm polyethylene glycol derivative is characterized in that the structure of the nitrogen-branched nine-arm polyethylene glycol derivative is shown as a general formula (1):
Figure FDA0002963906350000011
wherein N is a nitrogen atom trivalent branching center;
U 0 being a tetravalent radical, 3U's in the same molecule 0 The same; a trivalent branching center of nitrogen atoms and 3U 0 Together form a nine-valent branched structure;
L 2 is absent or L 2 Is a divalent linking group; in the same molecule, 9L 2 Are the same or different from each other;
PEG is a polyethylene glycol block taking ethylene oxide as a repeating unit, and both ends of the PEG are oxygen radicals;
F G is a hydrogen atom or a terminal functional group; when F is present G Containing at least one terminal functional group R when not a hydrogen atom 01 (ii) a Wherein R is 01 Is a functional group capable of interacting with biologically relevant substances.
2. The nitrogen-branched nine-arm polyethylene glycol derivative according to claim 1, wherein the PEG has the general formula
Figure FDA0002963906350000012
Wherein n is the polymerization degree of a polyethylene glycol chain and is selected from 1-2000; the polymerization degrees of the nine PEG chains may be the same as or different from each other, and are respectively represented by n 1 、n 2 、n 3 、n 4 、n 5 、n 6 、n 7 、n 8 、n 9 (ii) a The nitrogen-branched nine-arm polyethylene glycol has a single structureDispersivity or polydispersity.
3. The nitrogen-branched nine-arm polyethylene glycol derivative according to claim 1, wherein F is G Is- (L) 0 -G) g -(F) k Wherein g is 0 or 1; g is a terminal branching group selected from a trivalent or higher valent connecting group which connects the PEG chain segment with a terminal functional group; l is 0 Is a divalent linking group, connecting the PEG chain segment with the terminal branching group G; the integer k is the number of F in a single functionalized end and is selected from 1 or 2-250; f G Contains functional group, F has the structure of- (Z) 2 ) q -(Z 1 ) q1 -R 01 Wherein q and q1 are each independently 0 or 1, Z 1 、Z 2 Each independently is a divalent linking group, R 01 Is a functional group capable of interacting with a biologically relevant substance; f is a hydrogen atom and participates in forming a terminal functional group, namely hydroxyl, amino or sulfydryl;
when g is 0, k is 1, L 0 G is not present, F is not a hydrogen atom, hydroxyethyl, hydroxyl terminated PEG chain;
when G is 1, G is present, L 0 And k may be present or absent, and k is 2 to 250, where F is allowed to be a hydrogen atom.
4. The nitrogen-branched nine-arm polyethylene glycol derivative according to claim 1, wherein the nitrogen-branched nine-arm polyethylene glycol derivative is stably present or degradable; in the same molecule, U 0 、L 2 、L 0 、G、(Z 2 ) q -(Z 1 ) q1 Either, or the linking groups formed by either and adjacent groups, each independently, may be stable or degradable.
5. The nitrogen-branched nine-arm polyethylene glycol derivative according to claim 1, wherein the trivalent branching center of the nitrogen atom is derived from a residue of any one of the following trifunctional small molecules:
Figure FDA0002963906350000021
6. The nitrogen-branched nine-arm polyethylene glycol derivative according to claim 1, wherein U is 0 Comprises
Figure FDA0002963906350000022
Figure FDA0002963906350000023
Any of the tetravalent core structures; wherein the asterisk end in the tetravalent nuclear structure points to the trivalent branching center of the nitrogen atom, and the three non-asterisk ends point to three divalent linking groups L 2 ；
The U is 0 Further preferably, the compound has any one of the following structures:
Figure FDA0002963906350000024
Figure FDA0002963906350000031
Figure FDA0002963906350000032
wherein j is 1 Is 0 or 1, n i1 Is 0,1 or 2, n i2 Is 0,1, 2 or 3;
the U is 0 Further selected from any one of the following structures:
Figure FDA0002963906350000033
Figure FDA0002963906350000041
Figure FDA0002963906350000051
Figure FDA0002963906350000061
Figure FDA0002963906350000071
wherein the asterisk end of the quadrivalent structure points to the trivalent branching center of nitrogen atoms, and the three non-asterisk ends point to three divalent linking groups L 2 。
7. The nitrogen-branched nine-arm polyethylene glycol derivative according to claim 1, wherein each PEG chain corresponds to a number average molecular weight of 500,600,700,800,900,1000,1500,2000,2500,3000,3350,3500,4000,5000,5500,6000,6500,7000,7500,8000,8500,9000,9500,10000,11000,12000,13000,14000,15000,16000,17000,18000,19000 or 20000 Da.
8. The nitrogen-branched nine-arm polyethylene glycol derivative according to claim 1, wherein n is n 1 、n 2 、n 3 、n 4 、n 5 、n 6 、n 7 、n 8 、n 9 The corresponding PEG branched chains are all polydisperse, and n 1 ≈n 2 ≈n 3 ≈n 4 ≈n 5 ≈n 6 ≈n 7 ≈n 8 ≈n 9 (ii) a The number average degree of polymerization is preferably from 2 to about 1500; more preferably from 2 to about 1000; more preferably from 5 to about 500; more preferably from about 11 to about 500; more preferably from about 22 to about 500; more preferably from about 30 to about 250; more preferably 34 to about 150.
9. The nitrogen-branched nine-arm polyethylene glycol derivative according to claim 1, wherein the six PEG chains each have monodispersity, preferably n 1 ＝n 2 ＝n 3 ＝n 4 ＝n 5 ＝n 6 ＝n 7 ＝n 8 ＝n 9 (ii) a The number of EO units is preferably 2 to 70; more preferably 3 to 70; more preferably 3 to 50; more preferably 3 to 25; more preferably any of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 18, 20, 22, 24, 26, 27, 28, 29, 30, 32, 34, 36, 38, 40, 42, 44, 45, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 67, 68, 70.
10. The nitrogen-branched nine-arm polyethylene glycol derivative according to claim 1,
the R is 01 The interaction with the biologically relevant substance is selected from the group consisting of covalent bond formation, hydrogen bond formation, fluorescence, and targeting;
the R is 01 Preferably reactive groups, reactive group variants, functional groups with therapeutic targeting, fluorescent functional groups; the variant is selected from any one of a precursor of a reactive group, an active form thereof as a precursor, a substituted active form, a protected form, a deprotected form;
the R is 01 Any one of the functional groups of the A-type to the H-type or a variation thereof is preferable;
Class A: activated ester groups and analogous structures of activated ester groups; the active ester group is selected from a succinimide active ester group, a p-nitrobenzene active ester group, an o-nitrobenzene active ester group, a benzotriazole active ester group, a 1,3, 5-trichlorobenzene active ester group, a fluoro-phenyl active ester group and an imidazole active ester group; similar structures of the active ester group are selected from 2-thione-3-thiazolidine formate group, 2-thiothiothiazolidine-3-carboxylic acid ester group, 2-thiopyrrolidine-N-formate group, 2-thiobenzothiazole-N-formate group and 1-oxo-3-thiooxoisoindoline-N-formate group;
class B: sulfonate, sulfinate, sulfone, sulfoxide, 1, 3-disulfonyl-2-propylcarbonylphenyl, sulfone methacryl;
class C: hydroxylamino group, mercapto group, amino group, halogen atom, haloacetamido group, tetramethylpiperidinyloxy group, dioxopiperidinyloxy group, ammonium salt, hydrazine group, residue of disulfide, ester group, thioester group, carbonate group, thiocarbonate group, dithiocarbonate group, trithiocarbonate group, xanthate group, peroxythiocarbonate group, tetrathiodiester group, O-carbonylhydroxylamino group, amide group, imide group, hydrazide group, sulfonylhydrazine group, hydrazone group, imino group, enamine group, alkynylamine group, protected hydroxyl group or mercapto group, protected amino group; the amino group comprises a primary amino group and a secondary amino group; the protected hydroxyl or thiol group is preferably a carbamate, monothiocarbamate, dithiocarbamate; the protected amino group is preferably a carbamate, monothiocarbamate, dithiocarbamate;
Class D: carboxyl, sulfonic, sulfenic, hydroxamic, thiohydroxamic, xanthic, acid halide, sulfonyl chloride, aldehyde, glyoxal, acetal, hemiacetal, hydrated aldehyde, keto, ketal, hemiketal, ketal, hydrated keto, orthoacid, orthoester, cyanate, thiocyanate, isocyanurate, isothiocyanate, ester, oxycarbonylhalide, dihydrooxazolyl, thioaldehyde, thioketo, thioacetal, thioketone hydrate, thioketal, hemithioketal, thioester, dithiodiester, thiohemiacetal, monothiohydrate, dithiohydrate, thiohydrate, thiocarboxylic, urea, thiourea, guanidine and protonated forms thereof, amidino and protonated forms thereof, acid anhydride, squaric ester, Hemisquarylium, N-carbamoyl-3-imidazolyl, N-carbamoyl-3-methylimidazolium iodide, imidoyl, nitronyl, hydroxyimino, ureido, thioureido, pseudoureido; the dihydrooxazolyl group comprises oxazolinyl and isoxazolinyl; the thiocarboxylic acid group comprises a thiocarboxylic acid group and a dithiocarboxylic acid group;
Class E: maleimide group, acrylate group, N-acrylamide group, methacrylate group, N-methacrylamide group, protected maleimide group, maleamidyl group, 1,2, 4-triazoline-3, 5-diketone group, azo group, cyclic olefin group; the cycloalkenyl is preferably any one of cyclooctenyl, norbornenyl, 7-oxa-bicyclo [2.2.1] hept-5-en-2-yl, bicycloheptadiene/2, 5-norbornadiene and 7-oxabicycloheptadienyl;
class F: epoxy, alkenyl hydrocarbyl, alkynyl hydrocarbyl; the alkenyl group is preferably an ethenyl group or a propenyl group; the alkenyl hydrocarbon group is preferably an allyl group; the alkynyl group is preferably propynyl; the alkynyl hydrocarbyl is preferably propargyl;
the class of the signal is a class G,
class Ga: cycloalkyne, cycloalkyne heterocarbyl, conjugated diolefin, hybridized conjugated diolefin, 1,2,4, 5-tetrazinyl;
class Gb: azido, a nitrile oxide group, a cyanooxide group, cyano, isocyano, aldoximo, diazo, diazonium ion, azoxy, nitrilo imino, N-oxyaldoximo, tetrazolo, 4-acetyl-2-methoxy-5-nitrophenoxy and diazotized forms thereof; a functional group capable of undergoing a 1, 3-dipolar cycloaddition reaction;
Class H: hydroxyl, protected hydroxyl, siloxy, protected bishydroxy, trihydroxysilyl, protected trihydroxysilyl; the hydroxyl is selected from any one of alcoholic hydroxyl, phenolic hydroxyl, enol hydroxyl and hemiacetal hydroxyl.
11. The nitrogen-branched nine-arm polyethylene glycol derivative according to claim 10,
the R is 01 A functional group selected from any one of the following classes A to J, variations of classes A to H, functional derivatives of class I-class J; the variant is selected from any one of a precursor of a reactive group, an active form thereof as a precursor, a substituted active form, a protected form, a deprotected form:
class A:
Figure FDA0002963906350000081
or class B:
Figure FDA0002963906350000091
or class C:
Figure FDA0002963906350000092
or class D:
Figure FDA0002963906350000093
or class E:
Figure FDA0002963906350000094
Figure FDA0002963906350000101
or class F:
Figure FDA0002963906350000102
or class G:
class Ga:
Figure FDA0002963906350000103
or class Gb:
Figure FDA0002963906350000104
or class H:
Figure FDA0002963906350000105
Figure FDA0002963906350000111
or class I:
Figure FDA0002963906350000112
or class J:
Figure FDA0002963906350000113
wherein M is 5 Is a ring-forming atom selected from any one of carbon atom, nitrogen atom, phosphorus atom and silicon atom; m 5 The cyclic structure is a 3-50-membered ring, preferably a 3-32-membered ring, more preferably a 3-18-membered ring, and still more preferably a 5-18-membered ring; the cyclic structure is preferably any one of the following groups, a substituted form of any one, or a hybridized form of any one: cyclohexane, furanose ring, pyranose ring, benzene, tetrahydrofuran, pyrrolidine, thiazolidine, cyclohexene, tetrahydropyran, piperidine, 1, 4-dioxane, pyridine, pyridazine, pyrimidine, pyrazine, 1,3, 5-triazine, 1,4, 7-triazacyclononane, tripeptide, indene, indane, indole, isoindole, purine, naphthalene, dihydroanthracene, xanthene, thioxanthene, dihydrophenanthrene, 10, 11-dihydro-5H-dibenzo [ a, d ] s ]Cycloheptane, dibenzocycloheptene, 5-dibenzocycloheptenone, quinoline, isoquinoline, fluorene, carbazole, iminodibenzyl, naphthylene ring, dibenzocyclooctyne, aza-dibenzocyclooctyne;
wherein, Y 1 Is a leaving group attached to sulfonyl, sulfinyl, oxysulfonyl or oxysulfinyl, selected from any one of methyl, ethyl, n-propyl, isopropyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, vinyl, phenyl, benzyl, p-methylphenyl, 4- (trifluoromethoxy) phenyl, trifluoromethyl, 2,2, 2-trifluoroethyl;
wherein W is F, Cl, Br or I;
wherein, W 2 Is F, Cl, Br or I;
wherein, W 3 Is a leaving group selected from F, Cl, Br, I, PhS;
wherein, the first and the second end of the pipe are connected with each other,
Figure FDA0002963906350000121
each is a cyclic structure having a ring backbone containing a nitrogen atom, a azonium ion, a double bond, an azo, a triple bond, a disulfide bond, an anhydride, an imide, a diene, the cyclic structure being selected from a carbocycle, a heterocycle, a benzoheterocycle, a substituted carbocycle, a substituted heterocycle, or a substituted benzoheterocycle;
wherein M is a carbon atom, a nitrogen atom, a phosphorus atom or a silicon atom on the ring;
wherein M is 8 Is a carbon atom, a nitrogen atom, a phosphorus atom or a silicon atom located on the ring; m 8 The number of ring-forming atoms of the ring is 4-50; preferably 4-32; more preferably 5 to 32;
wherein M is 22 Is a carbon atom, a nitrogen atom, a phosphorus atom or a silicon atom on an alicyclic or alicyclic ring; m 22 The number of ring atoms of the ring is 4, 5, 6, 7 or 8;
wherein R is 2 Is a terminal group or a divalent linking group to which an oxygen or sulfur atom is bonded, selected from a hydrogen atom, R 21 Or R 3 Any one atom or group;
wherein R is 21 Is a divalent linking group and participates in ring formation; r 21 Is selected from C 1-20 Alkylene, divalent C 1-20 Heterohydrocarbyl, substituted C 1-20 Alkylene, substituted divalent C 1-20 Any divalent linking group or any two or any three of divalent linking groups in the heterohydrocarbon group; r 21 Preferably methylene, ethylene, propylene, butylene, pentylene, hexylene, heptylene, octylene, nonylene, decylene, 1, 2-phenylene, benzylene, C 1-20 Oxaalkylene, C 1-20 Thiaalkylene group, C 1-20 Any one of azaalkylene and azaaralkyl group, any one of themAny two or more of the same or different groups or combinations thereof in substituted form;
wherein R is 3 Is a terminal group linked to an oxy or thio group, selected from C 1-20 Hydrocarbyl radical, C 1-20 Heterohydrocarbyl radical, C 1-20 Substituted hydrocarbyl radical, C 1-20 Any of substituted heterohydrocarbyl groups; preferably any one or substituted form of any one of methyl, ethyl, n-propyl, isopropyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, benzyl, allyl;
wherein R is 4 Is- (R) 4 )C＝N + ＝N — Or- (R) 4 )C - -N + A hydrogen atom, a substituent atom or a substituent on C in the structure of [ identical to ] N; preferably any one atom or group of hydrogen atom, methyl group, ethyl group, n-propyl group, isopropyl group, butyl group, pentyl group, hexyl group, heptyl group, octyl group, nonyl group, decyl group, allyl group, propenyl group, vinyl group, phenyl group, methylphenyl group, butylphenyl group, benzyl group;
wherein R is 8 、R 9 、R 10 、R 11 、R 12 Each independently is a hydrogen atom, a substituent atom or a substituent on a double bond (-C-), and R is in the same molecule 8 、R 9 、R 10 、R 11 、R 12 May be the same as or different from each other; r 8 、R 9 、R 10 、R 11 、R 12 Each independently selected from: hydrogen atom, fluorine atom, methyl group; in class E3, R 8 Preferably methyl;
wherein R is 24 Is a terminal group linked to a disulfide bond selected from: c 1-20 Alkyl, aryl, hybrid phenyl;
wherein R is 27 Is a substituent attached to azo selected from: phenyl, substituted phenyl or hybrid phenyl;
wherein R is 30 Is a hydrocarbyl group selected from: c 1-20 Alkyl, benzyl, phenyl ring hydrogen atom by C 1-20 A hydrocarbyl-substituted benzyl group;
wherein M is 19 、M 20 、M 21 Each is independentThe sites are oxygen atoms or sulfur atoms, and may be the same or different from each other in the same molecule;
wherein, X 6 Is a terminal group attached to the oxygen atom of the ester group and is selected from a hydroxyl protecting group or the group LG 4 ；LG 4 Is selected from C 1-20 Alkyl, aryl, aralkyl, C 1-20 Heteroalkyl, heteroaryl, heteroaralkyl, C 1-20 Alkylcarbonyl, arylcarbonyl, aralkylcarbonyl, C 1-20 Heteroalkylcarbonyl, heteroarylcarbonyl, heteroaralkylcarbonyl, C 1-20 Alkoxycarbonyl, aryloxycarbonyl, aralkyloxycarbonyl, C 1-20 Alkylthio-carbonyl, arylthio-carbonyl, aralkylthiocarbonyl, C 1-20 Alkylaminocarbonyl, arylaminocarbonyl, aralkylaminocarbonyl, C 1-20 Heteroalkyloxycarbonyl, heteroaryloxycarbonyl, heteroaralkyloxycarbonyl, C 1-20 Heteroalkylthio-carbonyl, heteroarylthio-carbonyl, heteroaralkylthio-carbonyl, C 1-20 Heteroalkylaminocarbonyl, heteroarylaminocarbonyl, heteroarylalkylaminocarbonyl, C 1-20 Alkylthio, arylthio, aralkylthiocarbonyl, C 1-20 Heteroalkylthiocarbonyl, heteroarylthiocarbonyl, heteroarylalkylthiocarbonyl, C 1-20 Alkoxythiocarbonyl, aryloxylthiocarbonyl, aralkyloxythiocarbonyl, C 1-20 Alkylthio thiocarbonyl, arylthio thiocarbonyl, aralkylthio thiocarbonyl, C 1-20 Alkylaminothiocarbonyl, arylaminothiocarbonyl, aralkylaminothiocarbonyl, C 1-20 Heteroalkyloxythiocarbonyl, heteroaryloxythiocarbonyl, heteroarylalkoxythiocarbonyl, C 1-20 Heteroalkylthio thiocarbonyl, heteroarylthio thiocarbonyl, heteroarylalkylthio thiocarbonyl, C 1-20 (ii) any one of a heteroalkylaminothiocarbonyl, heteroarylaminothiocarbonyl or a substituted version of any one of the groups; wherein, the substituent atom or the substituent group is fluorine atom, alkoxy or nitro;
wherein, X 11 Is a terminal group attached to a carbonyl or thiocarbonyl group, selected from C 1-20 An alkyl group;
wherein, X 12 Is a terminal group to which a carbonate or thiocarbonate group is attached, selected from C 1-20 A hydrocarbyl group;
wherein, X 13 Is a terminal group for attaching a sulfur group selected from: mercapto-protecting group, group LG 2 ；
Wherein LG is 2 Selected from the group consisting of methyl, ethyl, n-propyl, isopropyl, t-butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, eicosyl, allyl, trityl, phenyl, benzyl, methylbenzyl, nitrobenzyl, t-butylthio, benzylthio, 2-pyridylthio, acetyl, benzoyl, methoxycarbonyl, ethoxycarbonyl, t-butyloxycarbonyl, phenoxycarbonyl, benzyloxycarbonyl, methylthiocarbonyl, ethylthiocarbonyl, t-butylthiocarbonyl, phenylthiocarbonyl, benzylthiocarbonyl, 2-pyridylcarbonyl, methylaminocarbonyl, ethylaminocarbonyl, t-butylaminocarbonyl, benzylamino-carbonyl, ethylthiocarbonyl, phenylmethylthiocarbonyl, methylthiocarbonyl, Methoxythiocarbonyl, ethoxythiocarbonyl, tert-butyloxythiocarbonyl, phenoxythiocarbonyl, benzyloxythiocarbonyl, methylthiothiocarbonyl, ethylthiothiocarbonyl, tert-butylthiothiocarbonyl, phenylthiothiocarbonyl, benzylthiocarbonyl, methylaminothiocarbonyl, ethylaminothiocarbonyl, tert-butylaminothiocarbonyl, benzylaminothiocarbonyl, C 1-10 Any one of a halogenated hydrocarbon group, a trifluoroacetyl group, a nitrophenyl group, or a substituted form of any one of the groups; wherein, the substituent atom or the substituent group is fluorine atom, alkoxy or nitro;
wherein Q is an atom or substituent contributing to the induction of unsaturated bond electrons, conjugation effect; when Q is on a ring, the number is one or more; when the number is multiple, the structure is the same, or the combination of two or more different structures; when a substituent group, Q has a linear structure, a branched structure containing a pendant group, or a cyclic structure;
wherein Q is 3 An atom or a group which is an H atom or a group contributing to the induction of electrons of unsaturated bonds, a conjugated effect, selected from any of a hydrogen atom, a fluorine atom, a chlorine atom, a bromine atom, an iodine atom, a methyl group, an ethyl group, an n-propyl group, an isopropyl group, a butyl group, a pentyl group, a hexyl group, a heptyl group, an octyl group, a nonyl group, a decyl group, a vinyl group, a propenyl group, an allyl group, a propynyl group, a propargyl group, a cyclopropyl group, a cyclopropenyl group, a phenyl group, a benzyl group, a butylphenyl group, a p-methylphenyl group, a p-nitrophenyl group, an o-nitrophenyl group, a p-methoxyphenyl group, an azaphenyl group, a methoxy group, an ethoxy group, a phenoxy group, a benzyloxy group, a methylthio group, an ethylthio group, a phenylthio group, a benzylthio group, a trifluoromethyl group, a 2, 2-trifluoroethyl group, or a substituted form of any of a group;
Wherein Q 5 Is H atom, methyl, ethyl or propyl; when Q is 5 When located on a ring, the number is one or more; when more than 1, the structure is the same, or the combination of two or more different structures;
wherein Q is 6 Is a hydrogen atom or a methyl group; q 7 Is a hydrogen atom, a methyl group, a phenyl group or a substituted phenyl group; in the same molecule, Q 6 And Q 7 May be the same or different;
wherein Q 8 Is a substituent atom or a substituent group on the imidazolyl group, and is selected from any one of H atoms, methyl groups, ethyl groups, propyl groups, butyl groups and phenyl groups; when Q is 8 Is one or more; when more than 1, the structure is the same, or the combination of two or more different structures;
wherein Q 11 Is a substituent on the nitrogen atom of the tetrazole, and is selected from any one of phenyl, substituted phenyl and aza-phenyl;
wherein PG 2 Is a thiol protecting group, the protected thiol group being denoted as SPG 2 Preferably any one of thioether, disulfide, silyl sulfide, thioester;
wherein PG 3 Is an alkynyl protecting group, preferably a silicon group;
wherein PG 4 Is a hydroxy protecting group, the protected hydroxy group being represented by OPG 4 Preferably ethers, silyl ethers, esters, carbonates, sulfonic acidsAny of esters;
wherein PG 5 Is an amino protecting group, the protected amino group being represented by NPG 5 Preferably any one of carbamate, amide, imide, N-alkylamine, N-arylamine, imine, enamine, imidazole, pyrrole and indole;
wherein PG 6 Is a bishydroxy protecting group, and PG 6 An acetal structure which forms a five-membered ring or a six-membered ring with two oxygen atoms; PG (Picture experts group) 6 Is methylene or substituted methylene; wherein PG 6 The substituent(s) is a hydrocarbyl substituent or a heteroatom-containing substituent selected from: any one of methylene, 1-methylmethylene, 1-dimethylmethylene, 1-cyclopentylene, 1-cyclohexylene, 1-phenylmethylene, and 3, 4-dimethylphenylmethylene;
wherein PG 8 Protecting groups for orthocarbonic acid or orthosilicic acid.
12. The nitrogen-branched nine-arm polyethylene glycol derivative according to claim 3,
said L 2 、L 0 (g＝1)、Z 1 、Z 2 Are all divalent linking groups, and are independent of each other, L in the same molecule 2 、L 0 (g＝1)、Z 1 、Z 2 May be the same as or different from each other;
L 2 、L 0 (g＝1)、Z 1 、Z 2 the structures of (a) are preferably each independently a linear structure, a branched structure or a cyclic-containing structure;
L 2 、L 0 (g＝1)、Z 1 、Z 2 each independently having 1 to 50 non-hydrogen atoms; more preferably 1 to 20 non-hydrogen atoms; more preferably 1 to 10 non-hydrogen atoms; the non-hydrogen atom is O, S, N, P, Si or B; when the number of the non-hydrogen atoms is more than 1, the types of the non-hydrogen atoms are 1, 2 or more than 2; when the number of the non-hydrogen atoms is more than 1, the carbon atoms and the carbon atoms, the carbon atoms and the heteroatoms, and the heteroatoms are combined;
L 2 、L 0 (g＝1)、Z 1 、Z 2 Either of which is selected so as to be able to,or any of the divalent linking groups with an adjacent heteroatom group are each independently a stably available linking group STAG or a degradable linking group DEGG; wherein the STAG can stably exist under any condition of light, heat, low temperature, enzyme, oxidation reduction, acidity, alkaline condition, physiological condition and in-vitro simulation environment; the DEGG is degradable under any one of light, heat, low temperature, enzyme, redox, acidity, alkalinity, physiological condition and in-vitro simulation environment;
the STAG is preferably an alkylene group, a divalent heteroalkyl group, a double bond, a triple bond, a divalent dienyl group, a divalent cycloalkyl group, a divalent cycloalkenyl group, a divalent cycloalkyne group, an aromatic ring group, an alicyclic ring group, a hetero heterocyclic group, a substituted alkylene group, a substituted divalent heteroalkyl group, a substituted double bond, a substituted dienyl group, a substituted divalent cycloalkyl group, a substituted divalent cycloalkenyl group, a substituted divalent cycloalkyne group, a substituted aromatic ring group, a substituted alicyclic ring group, a substituted hetero heterocyclic group, a substituted hetero benzene ring group, a substituted aromatic heterocyclic group, a substituted hetero heterocyclic group, an ether bond, a thioether bond, urea bond, thiourea bond, carbamate group, thiocarbamate group, -P (═ O) -, a divalent silicon group containing no active hydrogen, a divalent linking group containing a boron atom, a secondary amino group, a tertiary amino group, a carbonyl group, a thiocarbonyl group, an amide group, a thioamide group, Sulfonamide, enamine, triazolyl, 4, 5-dihydroisoxazolyl, and the like,
Figure FDA0002963906350000141
Any divalent connecting group in the skeleton of the amino acid and the derivative thereof, and a stable divalent connecting group formed by any two or more than two groups;
the DEGG preferably contains a disulfide bond, a vinyl ether bond, an ester group, a thioester group, a dithioester group, a carbonate group, a thiocarbonate group, a dithiocarbonate group, a thiocarbonate group, a carbamate group, a thiocarbamate group, a dithiocarbamate group, an acetal group, a cyclic acetal group, a mercaptide group, an azaacetal group, an azathiolacetal group, a dithioacetal group, a hemiacetal group, a thiohemiacetal group, an azahemiacetal group, a ketal group, a mercaptide group, an azaketal group, an azathioketal group, an imine bond, a hydrazone bond, an acylhydrazone bond, an oxime bond, a sulfoximine group, a semicarbazide bond, a semicarbazone bond, a thiocarbazone group, a hydrazide group, a thiocarbohydrazide group, an azocarbohydrazide group, a thioacylcarbonylhydrazide group, a hydrazinocarbohydrazide group, a carbazate group, a hydrazinothiocarbamate group, a, Carbazolyl, thiocarbazoyl, azo, isoureido, isothioureido, allophanate, thioallophanate, guanidino, amidino, aminoguanidino, aminoamidino, imidoyl, imidothioester, sulfonate, sulfinate, sulfonylhydrazino, sulfonylureido, maleimide, orthoester, benzyloxycarbonyl, phosphate, a phosphite group, a hypophosphite group, a phosphonate group, a phosphosilane group, a silane ester group, a carbonamide group, a thioamide group, a sulfonamide group, a polyamide group, a phosphorus amide group, a pyrophosphoric amide group, a cyclic phosphorus amide group, an isocyclic phosphorus amide group, a thiophosphoric amide group, an aconityl group, a polypeptide fragment, a nucleotide and derivative skeleton thereof, a deoxynucleotide and derivative skeleton thereof, and a combination of any two or more divalent linking groups.
13. The nitrogen-branched nine-arm polyethylene glycol derivative according to claim 3, wherein g ═ 1; the terminal branching groups G of the nitrogen-branched nine-arm polyethylene glycol derivative have the same structure type, and the structure type of the G is any one of branched, cyclic structure-containing, comb-shaped, tree-shaped and hyperbranched types; g may be degradable or may exist stably.
14. The nitrogen-branched nine-arm polyethylene glycol derivative according to claim 3, wherein G is 1, and G is selected from any one of the following structures:
when k is 2, G is a trivalent group; l is 0 -G comprises a structure selected from any one of the following groups: e 0 ,
Figure FDA0002963906350000151
Figure FDA0002963906350000152
Wherein, E is 0 Contains any one of the following structures:
Figure FDA0002963906350000153
Figure FDA0002963906350000154
Figure FDA0002963906350000155
wherein Q is 5 Is H atom, methyl, ethyl or propyl; when Q is 5 When located on a ring, the number is one or more; when more than 1, the structure is the same, or the combination of two or more different structures; wherein, the asterisk marks in the structure, which indicates that the asterisk end points to the polyethylene glycol unit;
said E 0 Preferably contains a capping structure selected from the group consisting of oxy, thio, secondary amino, divalent tertiary amino and 1, 2 or 3 of the same or different divalent linking groups in the carbonyl group;
said E 0 Any of the following structures is preferred:
Figure FDA0002963906350000161
Figure FDA0002963906350000162
Figure FDA0002963906350000171
Figure FDA0002963906350000172
Wherein, the asterisk marks in the structure, which indicates that the asterisk end points to the polyethylene glycol unit;
said E 0 Preferably a trivalent backbone structure of an amino acid or a derivative thereof; wherein the amino acid is L Is of type or D -type; amino acid orThe derivative is derived from any one of the following: serine, threonine, cysteine, tyrosine, hydroxyproline, aspartic acid, glutamic acid, asparagine, glutamine, lysine, arginine, citrulline, histidine, tryptophan.
15. The nitrogen-branched nine-arm polyethylene glycol derivative according to claim 3, wherein when k is 3, G is a tetravalent group; tetravalent G containing atoms CM 4 And unsaturated bond CB 4 Ring structure CC 4 Any tetravalent core structure, or comprises two trivalent core structures; l is 0 -G contains any of the following structures:
Figure FDA0002963906350000173
Figure FDA0002963906350000181
Figure FDA0002963906350000191
wherein X 1 Selected from any one of methyl, ethyl, n-propyl, isopropyl, tert-butyl, pentyl, hexyl, allyl, trityl, phenyl, benzyl, nitrobenzyl, p-methoxybenzyl and trifluoromethylbenzyl; wherein the asterisks within the structure indicate that the asterisk end points to the polyethylene glycol unit.
16. The nitrogen-branched nona-armed polyethylene glycol derivative according to claim 3, wherein when k is greater than or equal to 3, i.e. the valence state of G is greater than or equal to 4, the G with the valence of k +1 contains 1 k +1 nuclear structure, or is formed by directly connecting and combining 2-k-1 low-valence groups with the valence of 3-k or is formed by connecting and combining 1 or more than 1 divalent spacer L 10 Indirectly combining the components; the 3-k valent low-valent groups can be the same or different, and the valences can be the same or different; for a k +1 valent core structure, when k is more than or equal to 4 and contains a k +1 valent core structure, the k +1 valent core structure isA cyclic structure; when containing two or more L 10 When L is 10 May be the same as or different from each other; the direct or indirect combination of K +1(k is more than or equal to 4) G, the combination mode is selected from any one of a comb combination mode, a tree combination mode, a branching combination mode, a hyperbranched combination mode and a ring combination mode;
k is more than or equal to 4, and in the G with k +1 valence formed by direct or indirect combination, the combination mode is selected from any one of a comb combination mode, a tree combination mode, a branching combination mode, a hyperbranched combination mode and a ring combination mode;
wherein, in the tree-shaped composite structure, G is selected from any one of the following:
Figure FDA0002963906350000192
Figure FDA0002963906350000193
Figure FDA0002963906350000201
wherein d represents an algebra of a tree combination mode and is selected from 2, 3, 4, 5 or 6; wherein ng represents the algebra of the tree combination mode; wherein, the asterisk marks in the structure, which indicates that the asterisk end points to the polyethylene glycol unit;
wherein, the basic unit of the multivalent G forming the branched or hyperbranched combined structure is selected from trivalent G and tetravalent G and is a mixed combination of the multivalent G and the lower valence form thereof;
Wherein, the basic unit of the multivalent G forming the comb-shaped combined structure is trivalent G, tetravalent G or pentavalent G; the comb-like composite structure is formed by any basic unit selected from the following groups: polyglycerols, polypentaerythritol, substituted propylene oxides, substituted propylene oxide and carbon dioxide groups, acrylates and derivatives thereof, methacrylates and derivatives thereof, basic units containing acetal structures (e.g., (1 → 6) β -D glucopyranoside), amino acids and derivatives thereof containing hydroxyl or thio groups, acidic amino acids and derivatives thereof, basic amino acids and derivatives thereof, or acetalized glucans formed by D-glucopyranose units joined end to end by any one of β -1,6 glycosidic linkages, α -1,6 glycosidic linkages, β -1,4 glycosidic linkages, α -1,4 glycosidic linkages, β -1,3 glycosidic linkages, α -1,3 glycosidic linkages, and oxidized forms of the above multimers;
wherein, polyvalent G in cyclic combination is selected from: a residue of a cyclic peptide or a derivative thereof, a residue of a cyclic monosaccharide or a derivative thereof, a residue of a cyclic polysaccharide or a derivative thereof, a backbone of 1,4, 7-tri-tert-butoxycarbonyl-1, 4,7, 10-tetraazacyclododecane, a backbone of 2-hydroxymethylpiperidine-3, 4, 5-triol, a backbone of 6-amino-4- (hydroxymethyl) -4-cyclohexyl- [4H,5H ] -1,2, 3-triol.
17. A process for the preparation of a nitrogen-branched nine-armed polyethylene glycol derivative according to any of claims 1 to 16, characterized in that it involves the following steps:
step one, adopting nine hydroxyl small molecules containing nine hydroxyl groups
Figure FDA0002963906350000202
The initiator system of (1); wherein, deprotonation of nine bare hydroxyl groups forms nonaxonium anions
Figure FDA0002963906350000203
Is stable under anionic polymerization conditions;
initiating ethylene oxide polymerization;
step three, adding a proton source into the intermediate product system with nine polyethylene glycol chains obtained in the step two after the reaction is finished, so as to obtain hydroxyl-terminated nitrogen-branched nine-arm polyethylene glycol;
step four, functionalizing the tail end of the nitrogen-branched nine-arm polyethylene glycol to obtain the nitrogen-branched nine-arm polyethylene glycol derivative, wherein the terminal functionalization is selected from any one of terminal linear functionalization and terminal branched functionalization; when g is 0, terminal linear functionalization is performed, and when g is 1, terminal branched functionalization is performed.
18. The nitrogen-branched nine-arm poly of claim 17The preparation method of the ethylene glycol derivative is characterized in that the nonahydroxyl small molecule can be a trifunctional small molecule with 1 residue as a nitrogen atom trivalent branching center and 3 residues as U 0 The tetrafunctional micromolecules are obtained through coupling reaction; wherein the residue may be a tri-functional small molecule with a trivalent branching center for the nitrogen atom contains three identical functional groups; the residue may be U 0 The tetrafunctional small molecule contains three or four identical functional groups, when the residue can be U 0 When the tetrafunctional micromolecule only contains three same functional groups, the different functional group end is connected with the trifunctional micromolecule of which the residue can be a nitrogen atom trivalent branching center;
among them, a trifunctional small molecule in which the residue may be a trivalent branching center of a nitrogen atom is preferable
Figure FDA0002963906350000211
Figure FDA0002963906350000212
Any one of (1);
the residue may be U 0 The tetrafunctional small molecule of (a) is preferably
Figure FDA0002963906350000213
Figure FDA0002963906350000214
Any of the above.
19. The method for preparing nitrogen-branched nine-arm polyethylene glycol derivative according to claim 18, wherein the nine-hydroxy small molecule initiator IN- (OH) 9 Selected from any one of the following structures:
Figure FDA0002963906350000215
Figure FDA0002963906350000216
wherein n is i3 Is 0 or 1, R 1 Is a hydrogen atom or a methyl group.
20. A method for preparing a nitrogen-branched nine-arm polyethylene glycol derivative according to any one of claims 1 to 16, wherein the method involves 1 nine functionalized small molecule having nine identical functional groups
Figure FDA0002963906350000221
Reacting with 9 linear double-end functionalized PEG derivative bilPEG molecules in a coupling reaction process to obtain a nitrogen-branched nine-arm polyethylene glycol derivative; wherein bilPEG is monodisperse or polydisperse; wherein functional groups at both ends of bilPEG may be the same or different; wherein, F 1 Contains a reactive functional group capable of reacting with a terminal functional group in the biliPEG to form a divalent linking group L 2 ；
The functional group at the other end of the bilPEG is the same as or different from the target structure; when different, the method further comprises branching the nitrogen into nine-arm polyethylene glycol
Figure FDA0002963906350000222
The terminal functionalization process of (a), the terminal functionalization being selected from any one of terminal linear functionalization and terminal branched functionalization; when g is 0, terminal linear functionalization is performed, and when g is 1, terminal branched functionalization is performed.
21. The method for preparing the nitrogen-branched nine-arm polyethylene glycol derivative according to claim 20, wherein the nine-functionalized small molecule can be prepared by a tri-functionalized small molecule with 1 residue which can be a nitrogen atom trivalent branching center and 3 residues which can be U 0 The tetrafunctional micromolecules are obtained through coupling reaction; wherein the residue can be used as a trifunctional small molecule of a nitrogen atom trivalent branching center and contains three same functional groups; the residue may be U 0 The tetrafunctional small molecule of (2) then contains three or four identical functional groups, when the residue can beU 0 When the tetrafunctional micromolecule only contains three same functional groups, the different functional group end is connected with the trifunctional micromolecule of which the residue can be a nitrogen atom trivalent branching center;
Among them, the tri-functional small molecule in which the residue may be a nitrogen atom trivalent branching center is preferable
Figure FDA0002963906350000223
Figure FDA0002963906350000224
Figure FDA0002963906350000225
Any one of (1); the residue may be U 0 The tetrafunctional small molecule of (2) is preferably
Figure FDA0002963906350000226
Figure FDA0002963906350000227
Figure FDA0002963906350000231
Figure FDA0002963906350000232
Any of the above.
22. The method of preparing a nitrogen-branched nine-arm polyethylene glycol derivative of claim 21, wherein the nine-functionalized small molecule is selected from any one of the following structures: IN- (OH) 9 、
Figure FDA0002963906350000233
Figure FDA0002963906350000234
Figure FDA0002963906350000241
Figure FDA0002963906350000242
Wherein n is i3 Is 0 or 1, R 1 Is a hydrogen atom or a methyl group, the IN- (OH) 9 Is any one of the nine hydroxyl small molecules set forth in claim 19.
23. The nitrogen-branched nine-arm polyethylene glycol derivative-modified bio-related substance is characterized in that the structure of the nitrogen-branched nine-arm polyethylene glycol derivative-modified bio-related substance is shown as a general formula (2):
Figure FDA0002963906350000251
wherein N is a nitrogen atom trivalent branching center;
U 0 being a tetravalent radical, 3U's in the same molecule 0 The same; n and three U 0 Together form a nine-valent branched structure;
L 2 is absent or L 2 Is a divalent linking group; in the same molecule, nine L 2 Identical or different, preferably identical, from one another;
PEG is a polyethylene glycol block taking ethylene oxide as a repeating unit, and both ends of the PEG are oxygen radicals;
k is the number of EF in a single functionalized end and is selected from 1 or 2-250;
In a single functionalized PEG chain, g is 0 or 1; g is a terminal branching group selected from a trivalent or higher valent connecting group which connects the PEG chain segment with a terminal functional group; l is 0 Is a divalent linking group which connects the PEG chain segment with the terminal branching group G;
when g is 0, k is 1, L 0 G is not present;
when G is 1, G is present, L 0 May be present or absent, k is 2-250;
EF may be expressed as ED (structure is
Figure FDA0002963906350000252
) Or EF 1 (structure is
Figure FDA0002963906350000253
) And D is not equal to E 01 (ii) a Wherein q, q 1 Each independently is 0 or 1; z is a linear or branched member 1 、Z 2 Each independently is a divalent linking group; wherein D is a residue formed by reacting the modified biologically-relevant substance with the nitrogen-branched nine-arm polyethylene glycol derivative; e 01 Is selected from R 01 Protected R 01 Deprotected R 01 Or blocked R 01 ；R 01 Is a reactive group capable of reacting with a biologically relevant substance; l is a linking group formed after a reactive group in the nitrogen branched nine-arm polyethylene glycol derivative reacts with a biologically relevant substance; wherein the number of D at the end of one branch chain is marked by k D ，0≤k D K is not more than k, k of each branched chain in the same molecule D Each independently the same or different, and the sum of the numbers of D (N) in any one of the nitrogen-branched nine-arm polyethylene glycol derivative molecules D ) At least 1, preferably at least 9; when G is 1, G- (EF) k Can be expressed as
Figure FDA0002963906350000254
The biologically-relevant substance preferably has a plurality of reaction sites, and the same biologically-relevant substance and the same R 01 The residues D obtained by the reaction may be the same or different;
the biologically-relevant substance modified by the nitrogen-branched nine-arm polyethylene glycol derivative can exist stably or can be degraded; in the same molecule, U 0 、L 2 、L 0 、G、(Z 2 ) q -(Z 1 ) q1 、(Z 2 ) q Any of L, or any of the linkages formed with adjacent groups, is independently of each other stably present or degradable.
24. The nitrogen-branched nine-arm polyethylene glycol derivative-modified bio-related substance according to claim 23,
when g is 0, the structure is shown as formula (3):
Figure FDA0002963906350000261
or when the g is 1 and the content of D is 100%, the structure is shown as the formula (4):
Figure FDA0002963906350000262
25. the nitrogen-branched nine-arm polyethylene glycol derivative-modified bio-related substance according to claim 23,
k at the end of nine PEG chains in the same molecule D All satisfy k being more than or equal to 1 D K, i.e. at least one D is attached to each branch chain;
preferably, k at the end of nine PEG chains in the same molecule D All satisfy k, i.e. all of the terminal reactive sites in the nitrogen-branched nine-armed polyethylene glycol derivative molecule are each independently linked to a D.
26. The nitrogen-branched nine-arm polyethylene glycol derivative-modified bio-related substance of claim 23, wherein the average content of D in a single molecule is greater than 75%, equal to 75%, or less than 75%; preferably greater than about 80%, more preferably greater than about 85%, more preferably greater than about 90%, more preferably greater than about 94%, and most preferably equal to 100%.
27. The nitrogen-branched nine-armed polyethylene glycol derivative-modified biologically-relevant substance according to any one of claims 23 to 26, wherein the L corresponding to the nine PEG chain ends in the same molecule is not exactly the same or the L corresponding to the nine PEG chain ends is the same; the L is a straight chain structure, a branched chain structure or a cyclic structure;
the L is a divalent linking group or a trivalent linking group;
any one of L is independently stable or degradable, and the connecting group formed by L and the adjacent heteroatom group is stable or degradable; the conditions under which the difference can exist stably and be degraded are selected from any one of the following conditions: light, heat, enzymes, redox, acidic, basic, physiological conditions or in vitro simulated environments;
preferably the L at the ends of the nine PEG chains of the same molecule have the same stability, i.e. are all stably present or all degradable;
the L preferably contains any one of the following stably existing linking groups: ether linkages, thioether linkages, urea linkages, thiourea linkages, carbamate groups, thiocarbamate groups, secondary amino groups, tertiary amino groups, amide groups, imide groups, thioamide groups, sulfonamide groups, enamine groups, triazole groups, isoxazole groups, or degradable linking groups comprising any of the following: disulfide bond, vinyl ether bond, ester group, thioester group, dithioester group, carbonate group, thiocarbonate group, dithiocarbonate group, trithiocarbonate group, carbamate group, thiocarbamate group, dithiocarbamate group, acetal, cyclic acetal, mercaptal, azaacetal, azacyclic acetal, azathio acetal, dithioacetal, hemiacetal, thiohemiacetal, azahemiacetal, ketal, thioketal, azaketal, azathioketal, imine bond, hydrazone bond, acylhydrazone bond, oxime bond, sulfoximine ether group, semicarbazone bond, thiocarbazone bond, hydrazonohydrazone group, hydrazine group, hydrazide group, thiocarbhydrazide group, azocarbohydrazide group, thioazocarbonylacyl group, hydrazinoformate group, hydrazinothiocarbamate group, carbazide, thiocarbhydrazide, azo group, isoureido group, isothioureido group, thioureido group, and the like, Allophanate group, thioallophanate group, guanidino group, amidino group, aminoguanidino group, aminoamidino group, imido group, thiothiolester group, sulfonate group, sulfinate group, sulfonamide group, sulfonylhydrazide group, sulfonylurea group, maleimide group, orthoester group, phosphate group, phosphite group, hypophosphite group, phosphonate group, phosphosilane group, silane ester group, carbonamide, thioamide, phosphoramide, phosphoramidite, pyrophosphoro amide, cyclophosphamide, ifosfamide, thiophosphoramide, aconityl group, peptide bond, thioamide bond.
28. The nitrogen-branched nine-arm polyethylene glycol derivative-modified bio-related substance of claim 23, wherein the nitrogen-branched nine-arm polyethylene glycol derivative-modified bio-related substance meets any one of the following conditions:
(1) g-0, having stable nine-valent centers
Figure FDA0002963906350000271
stabilized-O- (Z) 2 ) q -L-；
(2) g-0, having stable nine-valent centers
Figure FDA0002963906350000272
degradable-O- (Z) 2 ) q -L-；
(3) g-0, with degradable nonavalent centers
Figure FDA0002963906350000273
degradable-O- (Z) 2 ) q -L-；
(4) g 1, having a stable nine-valent center
Figure FDA0002963906350000274
stabilized-O-L 0 -G- (G and Z are excluded) 2 Connection of), stable- (Z) 2 ) q -L- (containing G and Z) 2 The connection of (c);
(5) g 1, having a stable nine-valent center
Figure FDA0002963906350000275
degradable-O-L 0 -G- (G and Z are excluded) 2 Connection of), stable- (Z) 2 ) q -L- (containing G and Z) 2 The connection of (c);
(6) g 1, having a stable nine-valent center
Figure FDA0002963906350000276
stabilized-O-L 0 -G- (G and Z are excluded) 2 Linked) degradable- (Z) 2 ) q -L- (containing G and Z) 2 The connection of (c);
(7) g 1, having a degradable nine-valent center
Figure FDA0002963906350000277
stabilized-O-L 0 -G- (G and Z are excluded) 2 Connection of), stable- (Z) 2 ) q -L- (containing G and Z) 2 The connection of (c);
(8) g 1, having a degradable nine-valent center
Figure FDA0002963906350000278
degradable-O-L 0 -G- (not containing G and Z) 2 Connection of), stable- (Z) 2 ) q -L- (containing G and Z) 2 The connection of (c);
(9) g-1 with degradable nine-valent centers
Figure FDA0002963906350000279
stabilized-O-L 0 -G- (G and Z are excluded) 2 Linked) degradable- (Z) 2 ) q -L- (containing G and Z) 2 The connection of (c);
(10) g 1, having a stable nine-valent center
Figure FDA00029639063500002710
degradable-O-L 0 -G- (G and Z are excluded) 2 Linked) degradable- (Z) 2 ) q -L- (containing G and Z) 2 The connection of (c);
(11) g 1, having a degradable nine-valent center
Figure FDA00029639063500002711
degradable-O-L 0 -G- (G and Z are excluded) 2 Linked) degradable- (Z) 2 ) q -L- (containing G and Z) 2 The connection of (c);
(12) g-0, having stable nine-valent centers
Figure FDA00029639063500002712
stabilized-O- (Z) 2 ) q -, degradable L;
(13) g 1, having a stable nine-valent center
Figure FDA00029639063500002713
stabilized-O-L 0 -G-[(Z 2 ) q -] k A degradable L;
(14) g-0, having stable nine-valent centers
Figure FDA00029639063500002714
stabilized-O- (Z) 2 ) q Degradable L-D;
(15) g 1, having a stable nine-valent center
Figure FDA00029639063500002715
stabilized-O-L 0 -G-[(Z 2 ) q -] k And degradable L-D.
29. The nitrogen-branched nine-arm polyethylene glycol derivative-modified biologically-relevant substance according to claim 23, wherein the biologically-relevant substance is selected from any one of the following: drugs, proteins, polypeptides, oligopeptides, protein mimetics, fragments, enzymes, antigens, antibodies and fragments thereof, receptors, aptamers to gene-related substances, polysaccharides, proteoglycans, glycoproteins, lipid compounds, hormones, vitamins, vesicles, liposomes, dyes, fluorescent substances, targeting factors, cytokines, neurotransmitters, extracellular matrix substances, plant or animal extracts, viruses, vaccines, cells, micelles; any of the following is preferred: nucleic acids, steroids, phospholipids, glycolipids; any of the following is preferred: small molecule drugs, nucleosides, nucleotides, oligonucleotides, antisense oligonucleotides, polynucleotides, steroids;
The bio-related substance is preferably in any one of the following states: biologically relevant substances are any of themselves, dimers or multimers, partial subunits or fragments, precursors, activation states, derivatives, isomers, mutants, mimetics, polymorphs, pharmaceutically acceptable salts, fusion proteins, chemically modified substances, genetically recombinant substances, and agonists, activators, inhibitors, antagonists, modulators, receptors, ligands or ligands, antibodies and fragments thereof, substrates for acting enzymes or enzymes of any thereof;
the bio-related substance is preferably a bio-related substance, a modified bio-related substance, or a complex bio-related substance; the biologically-relevant substance is allowed to have a target molecule, an adjunct or a delivery vehicle bound thereto before or after binding to the nitrogen-branched nine-arm polyethylene glycol derivative to form a modified biologically-relevant substance or a complexed biologically-relevant substance;
the biologically-relevant substance is preferably any one of the following drugs: a medicament for treating any one of cancer, tumor, liver disease, diabetes, gout, rheumatism, rheumatoid, senile dementia, cardiovascular disease, antiallergic drug, anti-infective agent, antibiotic agent, antiviral agent, antifungal agent, central nervous system inhibitor, central nervous system stimulant, psychotropic drug, respiratory drug, peripheral nervous system drug, drug acting at synaptic or neuroeffector junction, smooth muscle active drug, histaminergic agent, antihistaminic agent, blood and hematopoietic drug, gastrointestinal drug, steroid agent, cell growth inhibitor, anthelmintic agent, antimalarial agent, antiprotozoal agent, antimicrobial agent, anti-inflammatory agent, immunosuppressive agent, Alzheimer's drug or compound, imaging agent, antidote, antispasmodic agent, muscle relaxant, anti-inflammatory agent, appetite suppressant, Migraine-treating agents, muscle contractants, antimalarials, antiemetics, bronchodilators, antithrombotic agents, antihypertensive agents, antiarrhythmic agents, antioxidants, antiasthmatic agents, diuretics, lipid regulating agents, antiandrogens, antiparasitic agents, anticoagulant agents, neoplastic agents, hypoglycemic agents, nutritional agents, food additives, growth supplements, anti-enteritis agents, vaccines, antibodies, diagnostic agents, contrast agents, hypnotic agents, sedatives, psychostimulants, tranquilizers, anti-parkinson agents, analgesics, anxiolytic agents, muscle infective agents, and auditory disease agents; the biologically-relevant substance is further preferably an anticancer or antitumor drug selected from anticancer or antitumor drugs for treating any one of the following diseases: breast cancer, ovarian cancer, cervical cancer, uterine cancer, endometrial cancer, gastrointestinal cancer, intestinal cancer, metastatic large intestinal cancer, rectal cancer, colon cancer, colorectal cancer, gastric cancer, squamous cell cancer, laryngeal cancer, esophageal cancer, malignant tumors, lung cancer, small-cell lung cancer, non-small cell lung cancer, liver cancer, thyroid cancer, renal cancer, bile duct cancer, brain cancer, skin cancer, pancreatic cancer, prostate cancer, bladder cancer, testicular cancer, nasopharyngeal cancer, head and neck cancer, gallbladder and bile duct cancer, retinal cancer, renal cell cancer, gallbladder adenocarcinoma, multidrug resistant cancer, melanoma, lymphoma, non-Hodgkin's lymphoma, adenoma, leukemia, chronic lymphocytic leukemia, multiple myeloma, brain tumor, Wilms' tumor, liposarcoma, endometrial sarcoma, rhabdomyosarcoma, neuroblastoma, and AIDS related cancers, Sarcoma or carcinosarcoma.
30. The nitrogen-branched nine-arm polyethylene glycol derivative-modified bio-related substance of claim 23, wherein the bio-related substance is preferably a small molecule drug selected from the group consisting of: biologically relevant substances with a molecular weight of not more than 1000Da and small molecular mimetics or active fragments of any of the biologically relevant substances; the small molecule drug is selected from any small molecule drug, or any derivative, or any pharmaceutically acceptable salt; the molecular weight of the small molecule drug is selected from any one of the following intervals: 0 to 300Da,300 to 350Da,350 to 400Da,400 to 450Da,450 to 500Da,500 to 550Da,550 to 600Da,600 to 650Da,650 to 700Da,700 to 750Da,750 to 800Da,800 to 850Da,850 to 900Da,900 to 950Da,950 to 1000 Da.
31. The nitrogen-branched nine-arm polyethylene glycol derivative-modified bio-related substance according to claim 23, wherein the small molecule drug is preferably selected from any one of flavonoids, terpenoids, carotenoids, saponins, steroids, quinones, anthraquinones, fluoroquinones, coumarins, alkaloids, porphyrins, polyphenols, macrolides, monobactams, phenylpropanoid phenols, anthracyclines, and aminoglycosides;
The small molecule medicament is preferably selected from any one of the following treatment fields: anti-cancer drugs, anti-tumor drugs, anti-hepatitis drugs, diabetes treatment drugs, anti-infective drugs, antibiotics, anti-viral agents, antifungal agents, vaccines, anti-respiratory drugs, anti-spasmodics, muscle relaxants, anti-inflammatory drugs, appetite suppressants, migraine agents, muscle contractants, antirheumatics, antimalarials, antiemetics, bronchodilators, antithrombotic agents, anti-hypertensive agents, cardiovascular agents, anti-arrhythmic agents, anti-oxidants, anti-asthmatic agents, diuretics, lipid regulators, anti-androgens, anti-parasitic agents, anticoagulants, neoplastic agents, hypoglycemic agents, nutritional agents and additives, growth supplements, anti-enteritis agents, vaccines, antibodies, diagnostic agents, contrast agents; preferably anticancer or antitumor drugs, antibiotics, antiviral agents, antifungal drugs, other anticancer drugs, antitumor drugs, antibiotics, antiviral agents, antifungal drugs; more preferably anticancer or antitumor drugs, antifungal drugs;
the small molecule drug is preferably any one of SN38, irinotecan, resveratrol, cantharidin and derivatives thereof, chrysin, tripterygium wilfordii extract, flavone or flavonoid drug, salvia miltiorrhiza extract and silybum marianum extract or any one of derivatives or any one of pharmaceutically acceptable salts; the pharmaceutically acceptable salt is preferably hydrochloride, oxalate, malate, citrate; most preferably a hydrochloride salt; the derivatives comprise molecular modified derivatives, glycosides, nucleosides, amino acids and polypeptide derivatives.
CN202110245443.7A 2021-03-05 2021-03-05 Nitrogen-branched nine-arm polyethylene glycol derivative, preparation method and modified biologically-relevant substance thereof Active CN115010915B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202110245443.7A CN115010915B (en) 2021-03-05 2021-03-05 Nitrogen-branched nine-arm polyethylene glycol derivative, preparation method and modified biologically-relevant substance thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202110245443.7A CN115010915B (en) 2021-03-05 2021-03-05 Nitrogen-branched nine-arm polyethylene glycol derivative, preparation method and modified biologically-relevant substance thereof

Publications (2)

Publication Number Publication Date
CN115010915A true CN115010915A (en) 2022-09-06
CN115010915B CN115010915B (en) 2024-06-11

Family

ID=83064835

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202110245443.7A Active CN115010915B (en) 2021-03-05 2021-03-05 Nitrogen-branched nine-arm polyethylene glycol derivative, preparation method and modified biologically-relevant substance thereof

Country Status (1)

Country Link
CN (1) CN115010915B (en)

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101831065A (en) * 2009-03-13 2010-09-15 复旦大学 Multi-arm star-shaped polyethylene glycol containing single-activity functional groups and preparation method thereof
CN102108119A (en) * 2009-12-25 2011-06-29 天津键凯科技有限公司 Multi-arm polyethylene glycol derivative, combination thereof with medicine, and gel
CN103044676A (en) * 2013-01-17 2013-04-17 厦门赛诺邦格生物科技有限公司 Polyethyleneglycol modified biologically-relevant substance
US20190117790A1 (en) * 2016-06-07 2019-04-25 Jenkem Technology Co., Ltd. (Beijing) Peg linker and ligand drug conjugate
CN110591079A (en) * 2018-06-13 2019-12-20 厦门赛诺邦格生物科技股份有限公司 Preparation method of monofunctional nonlinear polyethylene glycol
CN112694608A (en) * 2019-10-23 2021-04-23 厦门赛诺邦格生物科技股份有限公司 Six-arm polyethylene glycol derivative, preparation method and modified biologically-relevant substance

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101831065A (en) * 2009-03-13 2010-09-15 复旦大学 Multi-arm star-shaped polyethylene glycol containing single-activity functional groups and preparation method thereof
CN102108119A (en) * 2009-12-25 2011-06-29 天津键凯科技有限公司 Multi-arm polyethylene glycol derivative, combination thereof with medicine, and gel
CN103044676A (en) * 2013-01-17 2013-04-17 厦门赛诺邦格生物科技有限公司 Polyethyleneglycol modified biologically-relevant substance
US20190117790A1 (en) * 2016-06-07 2019-04-25 Jenkem Technology Co., Ltd. (Beijing) Peg linker and ligand drug conjugate
CN110591079A (en) * 2018-06-13 2019-12-20 厦门赛诺邦格生物科技股份有限公司 Preparation method of monofunctional nonlinear polyethylene glycol
CN112694608A (en) * 2019-10-23 2021-04-23 厦门赛诺邦格生物科技股份有限公司 Six-arm polyethylene glycol derivative, preparation method and modified biologically-relevant substance

Also Published As

Publication number Publication date
CN115010915B (en) 2024-06-11

Similar Documents

Publication Publication Date Title
US10660969B2 (en) Eight-arm polyethylene glycol derivative, production method therefor, and modified bio-related substance thereof
CN110643032B (en) Eight-arm polyethylene glycol, preparation method, functionalized derivative and modified biologically-relevant substance
EP1503802B1 (en) Controllably degradable polymeric biomolecule or drug carrier and method of synthesizing said carrier
CN110591079B (en) Preparation method of monofunctional nonlinear polyethylene glycol
Wang et al. Fabrication of zwitterionic and pH-responsive polyacetal dendrimers for anticancer drug delivery
US9180202B2 (en) Polyoxazoline polymers and methods for their preparation, conjugates of these polymers and medical uses thereof
US9186410B2 (en) Derivatized hyperbranched polyglycerols
Sedlacek et al. Poly (2-oxazoline)-protein conjugates
CN104109235A (en) Mono-functionalized polyethylene glycol with nitrogen atom branched center and its preparation method and biologically-relevant matter
CN107096038B (en) Preparation method of cross-linked nano-drug based on active reaction type one-step method
CN108530637B (en) Preparation method of single-functionalized branched polyethylene glycol
Liang et al. Fabrication of unimolecular micelle-based nanomedicines from hyperbranched polymers containing both terminal and internal reactive groups
CN113429559A (en) Six-arm polyethylene glycol derivative with end hetero-functionalized and preparation method thereof
US20170119898A1 (en) Hyperbranched polyglycerol sulfates with hydrophobic cores
Yao et al. PEGylated polylysine derived copolymers with reduction‐responsive side chains for anticancer drug delivery
CN108530636B (en) Single functionalized branched polyethylene glycol
US20080226739A1 (en) Hierarchically self-assembling linear-dendritic hybrid polymers for delivery of biologically active agents
CN115010915A (en) Nitrogen branched nine-arm polyethylene glycol derivative, preparation method and biological related substance modified by nitrogen branched nine-arm polyethylene glycol derivative
CN112694608B (en) Six-arm polyethylene glycol derivative, preparation method and modified biological related substance
Chen et al. Tumor extracellular pH-sensitive polymeric nanocarrier-grafted platinum (iv) prodrugs for improved intracellular delivery and cytosolic reductive-triggered release
Legros Engineering of poly (2-oxazoline) s for a potential use in biomedical applications
CN112694608A (en) Six-arm polyethylene glycol derivative, preparation method and modified biologically-relevant substance
Luo et al. Facile synthesis of high molecular weight poly (ethylene glycol)-b-poly (amino acid) s by relay polymerization
AU2015221541B2 (en) Derivatized hyperbranched polyglycerols

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant