CN112694608A

CN112694608A - Six-arm polyethylene glycol derivative, preparation method and modified biologically-relevant substance

Info

Publication number: CN112694608A
Application number: CN201911013365.7A
Authority: CN
Inventors: 翁文桂; 刘超; 王爱兰; 闫策
Original assignee: XIAMEN SINOPEG BIOTECH CO Ltd
Current assignee: XIAMEN SINOPEG BIOTECH CO Ltd
Priority date: 2019-10-23
Filing date: 2019-10-23
Publication date: 2021-04-23
Anticipated expiration: 2039-10-23

Abstract

The invention discloses a hexa-arm polyethylene glycol derivative shown in a general formula (1), a preparation method and a biological related substance modified by the hexa-arm polyethylene glycol derivative, wherein 1 trivalent central structure A₀And 3 symmetrical trivalent branched structures A₁Together forming a hexavalent central structure; l is₂Connection A₁And a polyethylene glycol arm; the polymerization degree of the six polyethylene glycol chains is n₁～n₆All selected from 1 to 2000; f_GIs- (L)₀‑G)_g‑(F)_k(ii) a g is 0 or 1, and a divalent linking group L can be arranged between the PEG chain and the F₀A terminal branched group G is connected, so that more reaction sites are provided, and the drug loading capacity is improved; k is the number of F in a single functionalized end and is selected from 1 or 2 to 250. The activity of the terminal hydroxyl of the hexahydroxy micromolecule initiator is basically the same, so that the performance of the hexa-arm polyethylene glycol derivative and the biological related substances modified by the hexa-arm polyethylene glycol derivative in large-scale production is more uniform and easy to control; degradable linking groups are allowed to exist in the linking groups, so that the drug can be degraded and released, and the drug effect is improved.

Description

Six-arm polyethylene glycol derivative, preparation method and modified biologically-relevant substance

Technical Field

The invention relates to the field of polymer synthesis, in particular to a six-arm polyethylene glycol derivative, a preparation method and a modified biologically-relevant substance.

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 general method for preparing monofunctional branched polyethylene glycols and their drug derivatives, and has been used in three commercially available 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, and the pharmaceutical activity can be kept high. In addition, the viscosity of the system 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 stages II and III.

Compared with four-arm polyethylene glycol, the six-arm polyethylene glycol can provide higher drug loading rate, better solubility and higher drug activity. The presently disclosed six-arm polyethylene glycol structure has dipentaerythritol ((-OCH) ₂)₃CCH₂C(CH₂O-)₃) And tetraglycerol (-OCH)₂(-O)CHCH₂OCH[CH₂OCH₂CH(O-)CH₂O-]₂) Two hexavalent central structures, but to our knowledge there are no cases of entry into preclinical studies or clinical stages.

Because the ether bond in the existing six-arm polyethylene glycol molecular structure is relatively stable, the polyethylene glycol molecule is not degradable, and because of the hexavalent central structure (-OCH) of the tetraglycerol branched polyethylene glycol₂(-O)CHCH₂OCH[CH₂OCH₂CH(O-)CH₂O-]₂) The polyethylene glycol has asymmetry, and two hydroxyl groups at each side of the terminal are asymmetric, so that the reactivity is different when ethylene oxide polymerization is initiated, the chain length of the polyethylene glycol is uneven, the molecular weight uniformity is not ideal, and the molecular weight distribution is wider.

Based on the background, it is necessary to develop a degradable initiator molecule with symmetrical branched terminal hydroxyl groups and basically the same activity, and further prepare a hexa-arm polyethylene glycol derivative with consistent chain length and uniform molecular weight.

Disclosure of Invention

The invention provides a six-arm polyethylene glycol derivative, a preparation method and a modified biological related substance against the background.

The purpose of the invention is realized by the following technical scheme:

a six-arm polyethylene glycol derivative has the following structural general formula:

wherein A is ₀Is a trivalent central structure;

A₁is a trivalent branched structure, three A₁Are all the same; each A₁Connection A₀And two L are led out₂And A is₁Middle two L₂The two ends of (a) are the same; a. the₀And A₁Covalently attached moieties, excluding A₀And A₁The respective part of the branched core is denoted L_A0A1，L_A0A1Is a divalent linking group containing an ether bond, a thioether bond, an ester bond, an amide bond, a carbonate bond, a urethane bond or a urea bond;

L₂is absent, or L₂To connect a trivalent branched structure A₁And a divalent linking group of PEG segments, six L₂Are all the same;

PEG has the general formula

Wherein one end is connected with L₂Is connected at the other end with F_GConnecting, wherein n is the polymerization degree of a polyethylene glycol chain and is selected from 1-2000; the degrees of polymerization of the six PEG chains, which may be the same or different from each other, are each represented by n₁、n₂、n₃、n₄、n₅、n₆(ii) a The six-arm polyethylene glycol has monodispersity or polydispersityDispersing; when monodisperse, the polydispersity PDI is 1. When polydisperse, the PDI>1, in which case the closer the PDI is to 1, the narrower the molecular weight distribution;

F_Gis- (L)₀-G)_g-(F)_kWherein 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 is₀Is 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-(Z₁)_q1-R₀₁Wherein q and q1 are each independently 0 or 1, Z₁、Z₂Each independently is a divalent linking group, R₀₁Is 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, L₀G is not present, F is not a hydrogen atom, hydroxyethyl, hydroxyl terminated PEG chain;

when G is 1, G is present, L₀May be present or absent, k is 2 to 250, where F is allowed to be a hydrogen atom;

the hexa-arm polyethylene glycol derivative can exist stably or can be degraded; in the same molecule, A₀、A₁、L₂、L₀、G、(Z₂)_q-(Z₁)_q1Each independently may be stable or degradable, and any of the above-described linkers to an adjacent group each independently may be stable or degradable.

The invention provides a preparation method of a six-arm polyethylene glycol derivative, which is prepared by the following technical scheme:

step one, adopting a hexahydroxy micromolecule containing six hydroxyl groups

The initiator system of (1); wherein deprotonation of six exposed hydroxyl groups forms the hexaoxide anions

(also note as

Or

) Is stable under anionic polymerization conditions;

initiating ethylene oxide polymerization;

step three, adding a proton source into the intermediate product system with six polyethylene glycol chains obtained in the step two after the reaction is finished, so as to obtain hydroxyl-terminated six-arm polyethylene glycol;

Step four, functionalizing the tail end of the hexa-arm polyethylene glycol to obtain a hexa-arm polyethylene glycol derivative, wherein the terminal functionalization is selected from any one of terminal linear functionalization and terminal branching 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 hexa-arm polyethylene glycol derivative, and the method relates to a hexa-functional small molecule containing six same functional groups

Reacting with six linear double-end functionalized PEG derivative bilPEG molecules in a coupling reaction process to obtain a six-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, F₁Contains a reactive functional group capable of reacting with a terminal functional group in the biliPEG to form a divalent linking group L₂；

The functional group at the other end of the bilPEG is the same as or different from the target structure; when different, the method also comprises the step of carrying out six-arm polyethylene glycol

The terminal functionalization process of (1), wherein the terminal functionalization is selected from any one of terminal linear functionalization and terminal branched functionalization(ii) a 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 six-arm polyethylene glycol derivative, which relates to 1 trifunctional micromolecule containing three same functional groups and 3 branched polyethylene glycol molecules V-PEG₂The coupling reaction process of (1), reacting to obtain a hexa-arm polyethylene glycol derivative; wherein, the branched V-PEG₂Containing two identical branched PEG chain segments and one end of a main chain functional group derived from a branched core, the functional groups of the two PEG chain ends and the main chain end may be the same or different, and V-PEG₂The main chain functional group end of the molecule is connected with the trifunctional micromolecule; wherein, V-PEG₂Is monodisperse or polydisperse;

the functional groups of the two PEG chain ends are the same or different from the target structure; when different, the method further comprises a process of end-functionalizing the hexa-armed polyethylene glycol derivative, wherein the end-functionalization is selected from any one of end-linear functionalization and end-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 biological related substance modified by the six-arm polyethylene glycol derivative, which has the following structural general formula:

wherein A is₀、A₁、L₂、PEG、g、G、L₀And k are as defined for formula (1) and are not described in detail herein.

₁Each independently is 0 or 1; z₁、Z₂Each independently is a divalent linking group; wherein D is a residue formed by the reaction of the modified bio-related substance and the hexa-arm polyethylene glycol derivative; e₀₁Is selected from R₀₁Protected R₀₁Deprotected R₀₁Or blocked R₀₁；R₀₁Is a reactive group capable of reacting with a biologically relevant substance; l is six-arm polyethylene glycolA linker formed after the reactive group in the alcohol derivative reacts with the bio-related substance; wherein the number of D at the end of one branch chain is marked by k_D，0≤k_DK is not more than k, k of each branched chain in the same molecule_DEach independently the same or different, and the sum of the numbers of D in any one of the hexa-armed polyethylene glycol derivative molecules (N)_D) At least 1, preferably at least 6; when G is 1, G- (EF)_kCan be expressed as

The biologically-relevant substance preferably has a plurality of reaction sites, and the same biologically-relevant substance and the same R₀₁The residues D obtained in the reaction may be the same or different.

The bio-related substance modified by the hexa-arm polyethylene glycol derivative can exist stably or can be degraded; in the same molecule, A₀、A₁、L₂、L₀、G、(Z₂)_q-(Z₁)_q1、(Z₂)_qEach L is independently, and any of the above-mentioned linking groups with adjacent groups are independently, either stably present or degradable.

Compared with the prior art, the invention has the following beneficial effects:

(1) the six-arm polyethylene glycol can be obtained by adopting symmetrical hexahydroxy micromolecules as initiator molecules through polymerization reaction. Compared with the prior six-arm polyethylene glycol product, the six-hydroxyl micromolecule can be used as A through one residue₀The trifunctional small molecule and 3 residues can be A₁The trifunctional micromolecules are obtained through coupling reaction. Wherein, the residue may be A₀The trifunctional small molecule of (a) contains three identical functional groups; the residue may be a₁The tri-functional small molecule of (A) contains two or three identical functional groups and is represented by₁At least two ends of the branched core are identical, when the residue can be A₁When the tri-functional small molecule contains only two identical functional groups, the different functional group ends and residues can be A₀Of (a) a trifunctional moietyThe sub-phases are connected. The six terminal hydroxyl groups of the small molecule with six hydroxyl groups are almost same in activity, and the six hydroxyl groups are respectively connected with A₁The six-arm polyethylene glycol derivative further prepared by taking the hexahydroxy micromolecule as an initiator molecule has the advantages of higher purity, more accurate control on molecular weight and distribution thereof in the product polymerization process, single product structure, no other multi-arm product structure, better performance, lower purification difficulty, reduced consumption of organic reagents in purification, lower cost, greener environmental protection and better product performance compared with the prior art in which the terminal hydroxyl asymmetric hexahydroxy micromolecule is taken as an initiator.

(2) Compared with linear polyethylene glycol, three-arm polyethylene glycol and four-arm polyethylene glycol, the six-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 six polyethylene glycol arms by direct initiation of ethylene oxide polymerization using a hexahydroxy small molecule initiator, six linear PEG arms can also be coupled to the end of a hexafunctional small molecule by a coupling method. The hexafunctional small molecule contains 6 identical functional groups selected from suitable reactive groups in class a-class H of the present invention. The hexafunctional small molecule passable residue may be A₀The three functional groups of the trifunctional micromolecule and the 3 residues can be A₁Is obtained by a coupling reaction of at least three functionalized small molecules containing the same functional group and is obtained from A₁At least two ends of the branched core are completely the same. When the residue may be A₁When the trifunctional small molecule of (a) contains only two identical functional groups, the different functional group or residue may be A ₀The activity of 6 terminal functional groups of the obtained hexafunctional micromolecule is almost the same, and the hexaarm polyethylene glycol can be endowed during the subsequent coupling reaction with the linear polyethylene glycolThe accurate molecular weight and narrow molecular weight distribution of the diol can reduce the difficulty of purification and separation, reduce the dosage of organic reagents, reduce the cost and be more 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 six-arm structure, a narrower molecular weight distribution can be achieved by reducing the PDI of the single-stranded starting material.

(5) Three branched polyethylene glycol molecules V-PEG containing two same branched PEG chain segments can also be combined by a coupling method₂Coupling to the end of a small trifunctional molecule with three identical functional groups results in a more uniform molecular weight of the coupled product.

(6) The product with single molecular weight of PDI-1 can be obtained, and the corresponding hexa-armed polyethylene glycol derivative and the bio-related substance modified by the hexa-armed polyethylene glycol derivative have definite structures and molecular weights, and have higher degree of standardized control and production.

(7) The six-arm polyethylene glycol derivative of the present invention, A₀、A₁、L₂、L₀、Z₁、Z₂Any one or any one of the linkers with an adjacent heteroatom allows for the presence of a degradable linker that can degrade under certain circumstances; when only A is₀When degradable, 1-3 two-arm branched V-shaped PEG can be degraded; when only L is present₂When degradable, 1-6 linear PEG chains can be degraded; when is- (Z)₂)_q-(Z₁)_q1When the position of (E) is degradable, 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 bio-related substance modified by the six-arm polyethylene glycol derivative, the functional group (F) and the polyethylene glycol chain allow the existence of a degradable connecting group, so that the biodegradable drug release is allowed under a specific environment, the tissue distribution of the drug is improved, the accumulation at a focus part is increased, and the drug effect is improved.

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 and various references, including but not limited to the term explanation 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 herein by reference 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 alkanes, 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, rings of hydrocarbon origin, 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, heterocycles, alicyclic heterocycles, aromatic heterocycles, heterocyclic rings, Heteroaromatic rings (heterocyclic-alicyclic rings), heteroaromatic rings (heterocyclic-aromatic rings), oxa-, aza-, thia-, phosph-, heteroatom numbers, oxa-, aza-, thia-, heteroatom positions, number of ring structures, single rings, multiple rings, single ring compounds, multiple ring compounds, number of rings, double rings, triple rings, four rings, linkages between ring structures, spiro rings, bridged rings, rings where any two of the multiple rings are linked, heteromonocyclic rings (monocyclic rings), hetero-cyclic rings, hetero-spiro rings, hetero-bridged rings, hetero-fused rings, fused aromatic rings (fused rings), fused heterocyclic rings (fused rings), fused aromatic rings (fused-heterocyclic rings), benzo-heterocyclic rings, fused aromatic rings, monocyclic hydrocarbons, spiro-cyclic hydrocarbons, bridged hydrocarbons, fused aromatic hydrocarbons, saturated cyclic hydrocarbons (aromatic hydrocarbons), fused aromatic hydrocarbons (fused cyclic hydrocarbons), fused aromatic hydrocarbons (aromatic hydrocarbons), heterocyclic hydrocarbons (fused heterocyclic hydrocarbons, fused heterocyclic hydrocarbons (fused heterocyclic hydrocarbons, fused aromatic hydrocarbons, fused heterocyclic hydrocarbons, unsaturated cyclic hydrocarbons, heterohydrocarbons, open-chain heterohydrocarbons, heterocyclic hydrocarbons, aliphatic heterohydrocarbons, aromatic heterohydrocarbons, aliphatic heterocyclic hydrocarbons, aliphatic heterocyclic open-chain hydrocarbons, saturated aliphatic heterohydrocarbons (heteroalkanes), heteroarenes, fused heterohydrocarbons, fused heterocyclic hydrocarbons, aromatic fused heterocyclic hydrocarbons, heteroaromatic alkanes and the like, and related structural examples and preferred modes.

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 group (alkyl), an unsaturated hydrocarbon group, an alkenyl group, an alkynyl group, a dienyl group, an alkenylhydrocarbon group, an alkynyl hydrocarbon group, a hydrocarbon group, an open-chain hydrocarbon, Unsaturated alicyclic hydrocarbon groups, monocyclic hydrocarbon groups, polycyclic hydrocarbon groups, aryl (aryl groups), aryl-hydrocarbon groups, aralkyl groups, heterohydrocarbon-substituted hydrocarbon groups (which may be heterohydrocarbon groups), aliphatic heterohydrocarbon groups, heteroalkyl groups, open-chain heterohydrocarbon groups, aliphatic heterohydrocarbon groups, heterocyclic hydrocarbon groups, heterocyclic-substituted hydrocarbon groups, heteroaromatic hydrocarbon groups, heteroaryl groups, heteroarylhydrocarbon groups, heteroaralkyl groups, fused cyclic hydrocarbon groups, fused aromatic groups, fused heterocyclic hydrocarbon groups, aromatic fused heterocyclic hydrocarbon groups, hetero-fused heterocyclic hydrocarbon groups, oxahydrocarbon groups, aza hydrocarbon groups, thiahydrocarbon groups, phosphane groups, mono-hetero heterohydrocarbon groups, bis-hetero hydrocarbon groups, hetero-hetero hydrocarbon groups, alkylene groups, sources of alkylene groups, alkylene groups derived from unsaturated aliphatic hydrocarbons, cycloalkylene groups, alicyclic hydrocarbon groups, arylene (arylene groups), arylenealkylene groups (aryl-hydrocarbene groups), cyclic structures as substituents, alkylene groups, may or may not contain substituents or pendant groups in the cyclic structures, The term "two positions in the alkylene group to which other groups are attached", protecting group, mercapto protecting group, alkynyl protecting group, hydroxyl protecting group, amino group, divalent linking group, etc., explanation of the relevant structures, and preferred modes. 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 hydrocarbon group formed by losing a hydrogen atom on a saturated carbon of the unsaturated hydrocarbon 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 where part other than the trivalent nuclear structure does not include a heteroatom, a trivalent group and its illustration where part other than the trivalent nuclear structure includes a heteroatom, a trivalent branching structure, a tetravalent group and its illustration where part other than the tetravalent nuclear structure does not include a heteroatom, a tetravalent group and its illustration where part other than the tetravalent nuclear 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 structural 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, exist stably, degradable divalent linking groups, degradable polyvalent groups, targeting factors, targeting molecules, photosensitive groups (covering fluorescent groups), fluorescent substances, heterofunctional group pairs (heterofunctional group pairs that can exist simultaneously), small molecules containing two same or different functional groups, small molecules, fluorescent substances, The definition, description, relevant examples and relevant citations of a 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, drug, application fields of drugs, application fields of small molecule drugs and the like. The preferred embodiments of the present invention are also included in the scope of the present invention.

In particular, CN201710126727.8 and the cited documents thereof are also incorporated into the present invention, as to the structure examples and preferred modes of protecting and deprotecting functional groups, particularly "protecting and deprotecting functional groups in reaction raw materials and intermediates". 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 amino carboxylic acids, protecting groups of natural amino acids, protecting and deprotecting methods of amino carboxylic 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. With respect to compounds, the group formed after a compound has lost part of the group is also referred to as a residue. 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. In particular, 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), and aryl groups (aryl hydrocarbon groups), respectively. Alkenyl, alkynyl, alkadienyl, aryl, and the like, with alkylene, constitute alkenylhydrocarbyl, alkynylalkyl, alkadienylalkyl, arylalkyl, and the like, while hydrocarbyl with alkenylene, alkynylene, alkadienylene, arylene, and the like, constitute respectively alkalkenyl, alkalkynyl, alkadienyl, alkarylyl, and the like. 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 end 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 into the category of aryl, 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 subscript position of C, indicating the number of carbon atoms the group has, e.g., C_1-10Denotes "having 1 to 10 carbon atoms", C_3-20Means "having 3 to 20 carbon atoms". ' get Substituted C_3-20Hydrocarbyl "means C_3-20A group obtained by substituting a hydrogen atom of a hydrocarbon group. "C_3-20The 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_1-10When 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 C₁、C₂、C₃、C₄、C₅、C₆、C₇、C₈、C₉、C₁₀Any of hydrocarbon groups.

The above definitions of integer numerical intervals composed of integers are not limited to subscripts of carbon atoms, but also apply to subscript intervals of other atoms, and also apply to integer numerical intervals in any subscript form or non-subscript form, including numerical intervals marked by short transverse lines (e.g., 1 to 6), and numerical intervals marked by wavy lines (e.g., 2 to 250), but not to numerical intervals composed 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, they are not required to be selected from the same preferred group of the same stage, but one may be selected from a wide range of preferences, one may be selected from a narrow range of preferences, one may be selected from a maximum range of preferences, and the other may be selected from any of the preferences, and the preferences in the same stage may be selected. For example, "R₈、R₉、R₁₀、R₁₁、R₁₂The number of carbon atoms of (A) is preferably 1 to 20, more preferably 1 to 201-10', may be 1-20, may be 1-10, may be partially 1-20, and the others are 1-10. Even if ranked or preferred by the same class, the structures of the two objects are not limited to be identical, for example A, B each independently selected from alkyl, cycloalkyl, aryl, aralkyl, and may be A is methyl and B is ethyl (both alkyl), or A is butyl and B is benzyl (one is alkyl and one is aralkyl).

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-CH₂CH₂-and-CH₂When an amide bond is used as a divalent linking group between-B, it may be A-CH ₂CH₂-C(＝O)NH-CH₂-B or A-CH₂CH₂-NHC(＝O)-CH₂-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 A₁Candidate of (2), the asterisk end points to trivalent center A₀Two identical non-star terminals each pointing to L₂E.g. A₁Is composed of

When the star ends are all pointing to the branching center A₀Two identical non-star terminals point to two L₂。

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 connected withWhen substituents contained in the linking group are easily confused, as in the structural formula

In (1), adopt

To mark the position of the divalent linking group to which the other group is attached, both of which represent-CH (CH) ₂CH₂CH₃)₂-、-CH₂CH₂CH(CH₃)₂-CH₂CH₂-. Or not particularly marked when no ambiguity arises, e.g. structural formulae marked as phenylene structures

Are respectively equivalent to

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,

represents an arbitrary cyclic structure;

represents an aliphatic cyclic structure and does not contain any aromatic or heteroaromatic ring, also known as an aliphatic ring;

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);

represents a skeleton of a saccharide or saccharide derivative having a cyclic monosaccharide skeleton, also referred to as a saccharide ring;

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;

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

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 carbocyclic rings of aliphatic hydrocarbon origin, being an all-carbon alicyclic ring. 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-dioxane. 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, and triazole is also included in the same 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 origin, being aromatic all-carbon rings. Aromatic-heterocyclic rings (aromatic-derived heterocyclic rings or aromatic heterocyclic rings)) refer to heterocyclic rings in which a carbon atom of a ring of the 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

Backbones of oligosaccharides or oligosaccharide derivatives

Polysaccharide or polysaccharide derivative backbone

Any of the above.

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 and the respective cited documents. 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 of isomer, chiral isomer, optical isomer, conformational isomer and rotamer, or a combination form of any two or more forms. 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, psicose, 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 linear, branched, hyperbranched, dendritic, comb-like or cyclic monosaccharide skeleton. The number of monosaccharide units is 2-10. Taking a cyclic mode as an example, any one of alpha-cyclodextrin, beta-cyclodextrin and gamma-cyclodextrin or derivatives thereof can be combined to form the cyclodextrin.

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, the D-glucopyranose units are linked in sequence by α -1,4 glycosidic linkages to form a linear combination; the linear structures are connected end to end, and 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 "hydrocarbon group" to 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.); also included, but not limited to, groups formed by directly linking a hydrocarbyl or heterohydrocarbyl group to a heteroatom-containing linker group such as oxy, thio, acyl, acyloxy, oxyacyl, -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. -CH₂-CH₂-CH₂-is replaced by-CH₂-S-CH(CH₃) - (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 also typically the shortest atomic distance, which can be used to indicate the length of the linking group; for example, in A-CO-NH-B the atomic separation of A and B is 2, A-p-Ph-CH ₂The atomic spacing of A and B in-B is 5(p-Ph is p-phenylene), and as another example A-CH (CH)₂CH₂CH₂CH₃) 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 a divalent linking group containing a cyclic structure, 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-CH₂–、–CH(CH₃)–、–C(CH₃)₂–、–CH(CH₂Ph)₂–、–C(CH₂OX) -with an atomic spacing of 1.

"carbon chain linkThe "linking group" refers to a linking group in which the main chain atoms are all carbon atoms, and the side chain moiety allows 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-CH₂-B、A-O-CH₂-B、

(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 one monovalent substituent, two monovalent substituents or one divalent substituent (e.g. divalent oxygen, e.g. together with the divalent methylene group to form a three-membered ring

) And (4) substitution. The substituted methylene group may be one in which a hydrogen atom is substituted (e.g., -CH (CH)₃) -) or two hydrogen atoms may be independently substituted (e.g., - (CH)₃)C(OCH₃) -, it being possible for two hydrogen atoms to be substituted simultaneously (for example carbonyl, thiocarbonyl, -C (-NH) -, -C (-N)⁺H₂) -) or may be cyclic side groups (e.g.

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. -CH₂-NH-CH₂-; if-C (═ O) -NH-is referred to as an amide bond, it is not considered to contain a secondary amino bond.

The number of corresponding repeating units in the "polymer chain" in the present invention is at least 2.

In the present invention, "molecular weight" represents the mass size of one molecule of a compound, and "average molecular weight" represents the mass size of a component of a compound of the formula in a macroscopic substance, and "average molecular weight" generally means "number average molecular weight" M _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 otherwise specified, the unit of measure of "molecular weight" and "average molecular weight" is 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 production method for obtaining a compound component having a set EO unit number, due to the limitations of the production method and the purification method, the macroscopic product may contain impurities of other EO unit number components than 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. gtoreq.10) or not more than. + -. 0.5 (base <10), it is considered that a monodispersed macroscopic product having the target component is obtained; furthermore, when the content of the component in accordance with the number of EO units or the range of 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 invention; even if the above content ratio is not achieved, the product having an insufficient content, the component in the form of a coproduct or a byproduct obtained therefrom, whether or not separation or purification is carried out, is within the scope of the present invention as long as the production method of the present invention or a similar method using substantially the same production concept is employed.

When describing the molecular weight of the compound formula of the polydisperse composition in Da, kDa, number of repeating units, number of EO units, the values fall within certain ranges of the given values (inclusive, preferably within ± 10%) for a single compound molecule; when the molecular weight of the compound formula describing the monodisperse component is a predetermined molecular weight, there is no fluctuation in the range, and it is a discrete point, 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 corresponding 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

Wherein one end is connected with L₂Is connected at the other end with F_GAnd 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 the breaking of the inventive chemical bond and the breaking into at least two residues independently of each other. 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 is not limited to, serum, heart, liver, spleen, lung, kidney, bone, muscle, fat, brain, lymph node, small intestine, gonad, etc., and may refer to intracellular, extracellular matrix, normal physiological tissue, and pathological 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. Degradable refers to 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 thiol group and 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, pathological 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, neutral, acidic, basic, physiological conditions, in vitro simulated environments, and the like, and preferably, light, heat, enzymes, redox, acidic, basic, and the like. The stable existence here means that stable linkage can be maintained in metabolic cycles in vivo without specific stimulation (e.g., pH condition at a specific site, light, heat, low temperature at the time of treatment, etc.) and molecular weight reduction due to chain cleavage (as long as integrity is maintained) does not occur.

In addition, the term "stably exist" with respect to the same linker is not an absolute concept, 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, an amide bond formed by dehydration condensation of an α -carboxyl group of a molecule of an amino acid and an α -amino group of a molecule of an amino acid, such as a peptide bond, can be cleaved when subjected to a particular enzyme, and is therefore also included in the "degradable" linker. Similarly, carbamate, thiocarbamate, and the like may be either 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 specific 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 more precisely the degradable nature of the compound structure, 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 referenced to the percent of dose that meets clinical evaluation criteria. For example, for an intravenously administered pegylated drug, when the blood concentration (based on the active pharmaceutical 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 a group that can exist stably in the present invention 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 component₂CH₂-O-CH₂CH₂Are 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 "not on or off" (on/off) comparison. 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 molecule 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 leaving group, etc., and 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, change of a leaving group and the like.

"reactive group variant" as used herein refers to a form in which a reactive group remains active (remains a reactive group) after at least one chemical change of oxidation, reduction, hydration, dehydration, electronic rearrangement, structural rearrangement, salt complexation and decomplexing, ionization, protonation, deprotonation, substitution, deprotection, change of leaving group, or the like, or an inactive form after it has been 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 and decomplexation, ionization, protonation, deprotonation, leaving group conversion, and the like. A transition of the leaving group, such as a transition from 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 means any two or any two of the aforementioned recited related structure typesCombinations of the above structures; 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-CH₂-Ph- (arylene-alkylene-arylene), -CH ₂-Ph-CH₂CH₂- (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-diazepine. 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 end₂And 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-L₅-COOH, wherein L₅Is 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 L₅Is alkylene (which may be open-chain or cyclic); further preferred is H₂N(CH₂)_j1COOH, wherein, the integer j₁The 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_LType-can also mean_DType-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 loss of a carboxylhydroxyl group (including all C-terminal carboxylhydroxyl groups, and also including carboxylhydroxyl groups on pendant 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 pendant 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 guanidino pendant 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, 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. The chemical modification products formed by modifying the reactive groups to change types, and 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 bind to it, 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 one molecule of the present invention is 2 or more, the same structure or general polymer formula is allowed without specific reference, and the molecular weight of the general polymer formula is allowed to be different. Such as Q, Q in the invention₃、Q₅、Q_eThe 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,Mercapto, alkylthio, and the like. A typical example of a lower alkyl group referred to in the present invention is C_1-6Alkyl, 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 amidinocarbonyl group, an amidinosulfonyl group, a formamido group, a sulfoxy group, a sulfonamide, a ureido 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.

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 Pointing to the inside of the ring and without other labels, representing a ring-forming atom attached at any suitable position, pointing to the outside and ending with a wavy line representing the free base end, which may constitute a covalent single bond with other groups.

Detailed Description

1. A hexa-arm polyethylene glycol derivative has a structure shown in a general formula (1):

wherein A is₀Is a trivalent central structure;

A₁is a trivalent branched structure, three A₁Are all the same; each A₁Connection A₀And two L are led out₂And A is₁Middle two L₂The two ends of (a) are the same;A₀and A₁Covalently attached moieties, excluding A₀And A₁The respective part of the branched core is denoted L_A0A1，L_A0A1Is a divalent linking group containing an ether bond, a thioether bond, an ester bond, an amide bond, a carbonate bond, a urethane bond or a urea bond;

L₂is absent, or L₂To connect a trivalent branched structure A ₁And a divalent linking group of PEG segments, six L₂Are all the same;

PEG has the general formula

Wherein one end is connected with L₂Is connected at the other end with F_GConnecting, wherein n is the polymerization degree of a polyethylene glycol chain and is selected from 1-2000; the degrees of polymerization of the six PEG chains, which may be the same or different from each other, are each represented by n₁、n₂、n₃、n₄、n₅、n₆(ii) a The six-arm polyethylene glycol has monodispersity or polydispersity;

F_Gis- (L)₀-G)_g-(F)_kWherein 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 is₀Is 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-(Z₁)_q1-R₀₁Wherein q and q1 are each independently 0 or 1, Z₁、Z₂Each independently is a divalent linking group, R₀₁Is 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;

1.1. Trivalent central structure A₀

Trivalent central structure A₀Is a trivalent group and contains a trivalent nuclear structure. The trivalent nucleus structure may be an atom CM₃One unsaturated bond CB₃Or a cyclic structure CC₃. Trivalent nuclear atom CM₃And a trivalent unsaturated bond nucleus structure CB₃Trivalent ring nucleus structure CC₃And preferred versions of the three include, but are not limited to, those described and exemplified in documents CN104877127A, CN104530413A, CN104530415A, CN104530417A and the respective cited documents. Taking CN104530417A as an example, corresponding to segment [0211]～[0284]. Taking CN104877127A as an example, the corresponding segment [0117 ]]～[0143]。

Wherein a trivalent nuclear atom CM₃There is no particular limitation as long as three covalent single bonds are allowed to be formed simultaneously. By way of example, trivalent nitrogen nuclei, trivalent carbon nuclei

Trivalent silicon nucleus

Trivalent phosphorus nuclei (e.g. of the formula

) And the like. The trivalent nuclear atom may be free of any atoms or groups, such as a trivalent nitrogen nucleus, or may be bound to other atoms or groups, such as a trivalent carbon nucleus, a trivalent silicon nucleus, a trivalent phosphorus nucleus, and the like.

R₃₇Substituents being the branching centre of trivalent silicon, selected from hydrocarbon radicals, preferably C_1-20Hydrocarbyl, more preferably C_1-20Alkyl, most preferably methyl.

R₁Is a hydrogen atom or a substituent attached to a carbon atom.

When taken as a substituent, R₁Are not particularly limited. Substituents which are stable under the conditions of anionic polymerization are preferred.

When taken as a substituent, R₁The 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, R₁May or may not contain heteroatoms.

When taken as a substituent, R₁The 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.

R₁Is a hydrogen atom or is selected from C_1-20Hydrocarbyl, substituted C_1-20A hydrocarbon group, etc. Wherein R is₁The 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 group.

R₁Preferably a hydrogen atom or C_1-20Alkyl, aralkyl, C _1-20Open-chain heterocarbyl, heteroaralkyl, substituted C_1-20Alkyl, substituted aryl, substituted C_1-20An open-chain heterocarbon group, a substituted heteroaromatic hydrocarbon group, or the like.

Specifically, as an example R₁Selected from a hydrogen atom or from the group comprising, 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_1-20Alkyl, substituted arylsHydrocarbyl, substituted C_1-20An 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_1-6Alkyl, alkoxy or nitro.

R₁Preferably 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, C group _1-10Halohydrocarbyl, haloacetyl or alkoxy substituted C_1-10An aliphatic hydrocarbon group. Wherein, the halogen atom is F, Cl, Br or I.

R₁Most preferably a hydrogen atom, a methyl group or an ethyl group.

Wherein, the trivalent unsaturated bond has a nuclear structure CB₃There is no particular limitation as long as three covalent single bonds can be formed simultaneously. The unsaturated bond may have two or more bonding atoms. Preferably 2 or 3. More preferably 2. By way of example, such as

And the like.

Wherein, the trivalent ring nucleus structure CC₃There is no particular limitation as long as three covalent single bonds can be simultaneously extracted. The ring-forming atoms from which the covalent single bond is derived are not particularly limited and include, but are not limited to, N, C, Si, P, and the like. The cyclic structure is selected from the group consisting of, but not limited to, aliphatic rings, aromatic rings, sugar rings, and condensed rings. The cyclic structure may be a single ring, such as a trivalent ring from cyclohexane, furanose, pyranose, benzene, pyridine, triazole, triazacyclononane, and the like; and may be polycyclic, such as rings from fluorene, carbazole, adamantane, and the like. Can be a naturally occurring cyclic structure, such as any trivalent monocyclic ring from any cyclic monosaccharide, or a trivalent ring generated by chemical reaction, such as cyclic peptide, lactone, lactam, lactide, etc. The covalent single bond may be directly derived from a ring-forming atom or may be derived from an unsaturated bond And (5) leading out. Three single covalent bonds may be drawn simultaneously from three ring-forming atoms, e.g.

Or wherein two single covalent bonds are from the same ring-forming atom

Wherein M is₅、M₆、M₇、M₂₃Are ring-forming atoms, i.e. atoms located on a ring. M₅、M₆、M₇、M₂₃Each independently is a carbon atom or a heteroatom, and may be the same as or different from each other in the same molecule. M₅、M₆、M₇、M₂₃Each independently is preferably a carbon atom, a nitrogen atom, a phosphorus atom or a silicon atom. M₅、M₆、M₇、M₂₃The ring is a 3-to 50-membered ring, preferably a 3-to 32-membered ring, more preferably a 3-to 18-membered ring, and still more preferably a 5-to 18-membered ring.

M₂₃Carbon atom, nitrogen atom, phosphorus atom or silicon atom with 2 single bonds led out from the ring. When a nitrogen atom, it is present in the form of a quaternary ammonium cation.

M₅、M₆、M₇Ring in which M is located₂₃、M₆The ring is not particularly limited, including but not limited to

And the like. The number of ring-forming atoms is not particularly limited, but is preferably 3 to 50-membered rings, more preferably 3 to 32, and still more preferably 3 to 18.

Wherein, the aliphatic ring

Is any alicyclic or alicyclic ring, and the ring-forming atoms are each independently a carbon atom or a heteroatom; the hetero atomThe atom is not particularly limited, and includes, but is not limited to, a nitrogen atom, an oxygen atom, a sulfur atom, a phosphorus atom, a silicon atom, a boron atom, and the like. The hydrogen atom on the ring-forming atom of the alicyclic ring may be substituted with any substituent atom or substituent, or may be unsubstituted. The substituted heteroatom or substituent is not particularly limited and includes, but is not limited to, any substituted heteroatom or any substituent of the term moiety selected from any of a halogen atom, a hydrocarbyl substituent, and a heteroatom-containing substituent. Broadly, the alicyclic and alicyclic rings include, but are not limited to, any one of the ring structures or any combination of two or more of the ring types in monocyclic, polycyclic, spiro, bridged, fused, carbocyclic, heterocyclic, alicyclic, heteromonocyclic, heteromulticyclic, heterospiro, heterobridged, heteroalicyclic.

Wherein the aromatic ring

Is any aromatic ring or aromatic heterocyclic ring, and the ring-forming atoms are each independently carbon atoms or heteroatoms; the hetero atom is not particularly limited and includes, but is not limited to, a nitrogen atom, a phosphorus atom, a silicon atom, a boron atom, and the like. The hydrogen atom on the ring-forming atom of the aromatic ring may be substituted with any substituent atom or any substituent, or may be unsubstituted. The substituted heteroatom or substituent is not particularly limited and includes, but is not limited to, any substituted heteroatom or any substituent of the term moiety selected from any of a halogen atom, a hydrocarbyl substituent, and a heteroatom-containing substituent. The substituent atom is preferably a halogen atom. The substituent is preferably a group that contributes to the induction, conjugation effect of the unsaturated bond electrons. Broadly, the aromatic rings and aromatic heterocycles: including, but not limited to, any one of the ring structures or any combination of two or more of the ring types monocyclic, polycyclic, fused ring, fused aromatic ring, fused heteroaromatic ring, carbocyclic ring, heterocyclic ring, aromatic heterocyclic ring, hetero-monocyclic, hetero-polycyclic, hetero-fused ring, and hetero-aromatic ring. The aromatic ring is preferably selected from the group consisting of benzene, pyridine, pyridazine, pyrimidine, pyrazine, 1,3, 5-triazine, tetrazine (1,2,3,4-, 1,2,4, 5-and 1,2,3, 5-isomers), indene, indane, indole, isoindole, purine, pyrimidine, pyrazine, triazine, Naphthalene, dihydroanthracene, xanthene, thioxanthene, dihydrophenanthrene, 10, 11-dihydro-5H-dibenzo [ a, d ]]Cycloheptane, dibenzocycloheptene, 5-dibenzocycloheptenone, quinoline, isoquinoline, fluorene, carbazole, iminodibenzyl, naphthylethyl, dibenzocyclooctyne, azabenzocyclooctyne, and the like, substituted versions of any, or hybridized versions of any.

Wherein, the sugar ring

Is a skeleton of a saccharide or a saccharide derivative having a cyclic monosaccharide skeleton. The saccharide or saccharide derivative is derived from natural monosaccharide or unnatural monosaccharide. The structure of the cyclic monosaccharide is any one form or a combination form of any two or more than two of an isomer, a chiral isomer, an optical isomer, a conformational isomer and a rotational isomer of the cyclic monosaccharide. For example, the pyranose ring may be in either the boat or chair conformation.

A backbone selected from a cyclic monosaccharide or a derivative of a cyclic monosaccharide

Backbones of oligosaccharides or oligosaccharide derivatives

Polysaccharide or polysaccharide derivative backbone

Any of the above.

And preferred versions of the three include, but are not limited to, those described and exemplified in documents CN104877127A, CN104530413A, CN104530415A, CN104530417A and the respective cited documents. 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, psicose, fructose, sorbose, tagatose, inositol. The cyclic structure is preferably a five-membered ring or a six-membered ring.

Wherein, condensed ring

Containing chemical bonds formed by condensation of amide bonds, ester bonds, imides, anhydrides, or the likeAnd (4) a ring. Examples are lactones, lactides, lactams, cyclic imides, cyclic anhydrides, cyclic peptides, etc.

CC₃The trivalent ring structure of (A) is preferably selected from the group consisting of cyclohexane, furanose, pyranose, benzene, tetrahydrofuran, pyrrolidine, thiazolidine, cyclohexene, tetrahydropyran, piperidine, 1, 4-dioxane, pyridine, pyridazine, pyrimidine, pyrazine, 1,3, 5-triazine, 1,4, 7-triazacyclononane, cyclic tripeptide, indene, indane, indole, isoindole, purine, naphthalene, dihydroanthracene, xanthene (e.g. xanthene), thioxanthene, dihydrophenanthrene, 10, 11-dihydro-5H-dibenzo [ a, d-dihydrodibenzo [ a, d ] and mixtures thereof]Cycloheptane, dibenzocycloheptene, 5-dibenzocycloheptenone, quinoline, isoquinoline, fluorene, carbazole, iminodibenzyl, naphthylethyl, dibenzocyclooctyne, azabenzocyclooctyne, and the like, substituted versions of any, or hybridized versions of any.

A₀The specific structure of (a) is exemplified as follows:

etc.; wherein Q is₅Is H atom, methyl, ethyl or propyl; when Q is₅When 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 integer j ₂Selected from any one of 0, 1, 2, 3, 4, 5, 6.

A₀Further preferred are structures containing the above structure terminated with 1, 2 or 3 divalent linking groups selected from the group consisting of oxy, thio, secondary amino, divalent tertiary amino and carbonyl groups, which may be the same or different. Wherein Q is₅Selected from a hydrogen atom, a substituent atom or a substituent group, is not particularly limited, and is preferably selected from a H atom, a methyl group, an ethyl group or a propyl group. When Q is₅When located on a ring, 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. Q₅The ring includes but is not limited toFluorene, carbazole, norbornene, 7-oxa-bicyclo [2.2.1]Hept-5-en-2-yl. By way of example, A₀May be selected from any of the following structures:

and the like. Wherein Q is₅The definitions of (a) and (b) are consistent with the above.

1.2. Trivalent branched structure A₁

A₁Is a trivalent branched structure, excluding the branching central atom or central branching ring, A₁Contains two identical end groups, or three identical end groups. A. the₁Contains a trivalent nucleus structure. The trivalent nucleus structure may be an atom CM₃One unsaturated bond CB₃Or a cyclic structure CC₃. Trivalent nuclear atom CM₃And a trivalent unsaturated bond nucleus structure CB ₃Trivalent ring nucleus structure CC₃And preferred versions of the three include, but are not limited to, those described and exemplified in documents CN104877127A, CN104530413A, CN104530415A, CN104530417A and the respective cited documents. Taking CN104530417A as an example, corresponding to segment [0211]～[0284]. Taking CN104877127A as an example, the corresponding segment [0117 ]]～[0143]. Wherein a trivalent nuclear atom CM₃And a trivalent unsaturated bond nucleus structure CB₃Trivalent ring nucleus structure CC₃The definition of (A) and the preferable conditions of the three are as described above₀The above description is not repeated here.

A₁Comprises

Any one of the trivalent nuclear structures; wherein A is₁The asterisk marks in the structure indicate that the asterisk ends point towards the trivalent center A₀Two non-star ends are the same and point to two L₂；

Wherein R is₁、R₃₇、M₅、M₆、M₇、M₁₉、M₂₃The definition of (A) and₀the above description is not repeated here.

A₁Specific examples of the structures, A₁Contains any one of the following structures:

wherein Q is₅Is H atom, methyl, ethyl or propyl; when Q is₅When 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 A is₁Asterisks in the structure indicate that the asterisk ends point towards the trivalent center A₀With two identical non-star terminals pointing to two L₂。

A₁Further preferred are compounds containing the above structure terminated with 1, 2 or 3 identical or different divalent linking groups selected from oxy, thio, secondary amino, divalent tertiary amino and carbonyl groups; when participating in initiator molecules constituting living anionic polymerization, no carbonyl group, secondary amino group.

A₁Further preferably, any of the following structures:

1.3. divalent linking group L_A0A1

A₀And A₁Covalently attached moieties, excluding A₀And A₁The respective part of the branched core is denoted L_A0A1，L_A0A1Is 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-). "A" is₀With a branched core of₁The atomic distance between the branched cores of (2) is greater than or equal to 1, preferably 1 to 100, that is, any atomic distance between 1, 2, 3, 4, 5, … …, 98, 99 and 100 or any atomic distance between 1, 1 to 10, 10 to 20, 20 to 50 and 50 to 100 can be arbitrarily selected.

Said "A" is₀By branched core "is meant the trivalent radical A₀The simplest trivalent moiety of (A) is removed₁A divalent linking group therebetween; said "A" is₁By branched core "is meant the trivalent radical A₁The simplest trivalent moiety of (A) is removed₀The divalent linking group in between, and L is also removed₂A linker group therebetween; "A" is₀Branched core of "," A₁Any of the base atoms at the free ends of the branched cores "is indispensable for constituting a trivalent structure, and together constitutes a trivalent A₀Trivalent A₁The absence of any free-end atom in the outermost periphery of the core skeleton of (a) results in a reduction in valence state or an incomplete structure.

Said "A" is₀With a branched core of₁The atomic distance between the branched nuclei of (A)'₀Branched core distance A of₁The atomic spacing between the branched nuclei of (a).

“A₀Branched core of "," A₁Judgment of branched core of (1) "exemplifies:

a in example 5₁

A in example 7₁

A in example 8₁

Etc. are all branched nuclei>CH-；

A in example 6₀

Etc. are all branched nuclei>C(CH₃)-；

Etc. are all branched nuclei>C(CH₂CH₃)-；

A in example 7₀

Etc. are all branched nuclei>Si(CH₃) -; a in example 5₀

A in example 6₁

A in example 9₀

A in example 22₀

A in example 23₁

Etc. are all branched nuclei of

Example 8A₀

Has a branched core of

Example 23A₀

Has a branched core of

L_A0A1When A is an example₀Is composed of

A₁Is composed of

When L is_A0A1is-CH₂OCH₂CH₂-, containing an ether bond; when A is₀Is composed of

A₁Is composed of

When L is_A0A1is-CH₂CH₂NHC(O)CH₂-, containing an amide bond; when A is₀Is composed of

A₁Is composed of

When L is_A0A1is-CH₂C (O) O-, containing an ester bond.

1.4. Divalent linking group L₂、L₀(g＝1)、Z₁、Z₂

L₂Is absent, or L₂To connect a trivalent branched structure A₁And a divalent linking group of PEG segments, six L₂Are all the same.

L₂Are groups that can be stably present or degradable. When L is₂In the case of a group which can be present stably, L₂Preferred 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 _1-20Alkyl, substituted aryl, substituted C_1-20An 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_1-6Alkyl, alkoxy or nitro.

L in the above general formula (1)₀When present, L is not a hydrogen atom₂、L₀(g＝1)、Z₁、Z₂Are 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.

L₂、L₀(g＝1)、Z₁、Z₂The 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.

L₂、L₀(g＝1)、Z₁、Z₂The 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; more preferably 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. Not being hydrogen atomsWhen the number is 1, the non-hydrogen atom may be a carbon atom or a hetero atom. 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.

L₂、L₀(g＝1)、Z₁、Z₂Each 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.

L₂、L₀(g＝1)、(Z₂)_q-(Z₁)_q1Is not particularly limited, and 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 existing linking group STAG or a degradable linking group DEGG. For the preferred case, L₀(g＝1)、(Z₂)_q-(Z₁)_q1Any 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), the polymerization degrees of six PEG chains may be the same as or different from each other, and may be represented by n₁、n₂、n₃、n₄、n₅、n₆. It is permissible that the number of EO units of the six PEG chains in the same molecule be the same as or different from each other, and that the degrees of polymerization of the six PEG chains in the macropolymer be the same as or different from each other. The six-arm polyethylene glycol can be polydisperse or monodisperse. Wherein, the PEG chain n _i(i ═ 1,2,3,4,5,6) can be both monodisperse and polydisperse, or any combination of monodisperse and polydisperse, preferably both monodisperse and polydisperse.

The polymerization mode is adopted to obtain six polyethylene glycol chains with the same polydispersity.

The PDI of the six-armed polyethylene glycol obtained by coupling depends on the polydisperse nature of the starting material, preferably the six 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_nIt may be either a polydisperse block or a molecular weight of a substance or a monodisperse block or a molecular weight of a substance, and unless otherwise specified, a polydisperse polymer is generally specified. When not specifically written, the units are daltons, Da.

PEG chain n for polydispersity_i(i ═ 1,2,3,4,5or 6), 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 preferable the above, the more conventional the molecular weight of the corresponding PEG segment is, the simpler and easier the preparation, and the narrower the PDI (polydispersity index) of the molecular weight is, the more uniform the performance is. The number average molecular weight of linear PEG obtained by a common polymerization method is about 2kDa to 40kDa, and in the present invention, the number average molecular weight of each PEG of the six-arm polyethylene glycol derivative also prepared by a polymerization method is selected from 2kDa to 40 kDa. In 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 monodisperse PEG blocks, the molecular weight is defined by the number of oxyethylene units (described 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 six-armed polyethylene glycol of formula (1) is determined collectively by the combination of the six PEG chains, and the six-armed polyethylene glycol species may be a single component or a mixture of different components, as long as the PDI of the polymer is 1. When a single component, the six PEG chains have the same number of EO units. When a mixture of different components, the overall molecular weight of each six-armed polyethylene glycol molecule in the polymer is fixed, but where the number of EO units of the six PEG chains 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, the six 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.

The polydispersity index for the entire six-arm polyethylene glycol derivative may be the same or different than the polydispersity of the individual PEG chains. But the lower the PDI of the whole compound, the better. Thus, for the six-arm polyethylene glycol derivative represented by the general formula (1)The chain length distribution of six PEG chains of the substance, preferably n₁≈n₂≈n₃≈n₄≈n₅≈n₆(in which case the number average molecular weights of the six chains may each independently be the same or close together) or n₁＝n₂＝n₃＝n₄＝n₅＝n₆(six chains now have fixed molecular weights and are equal to each other). The chain lengths of the PEG chains are equal or similar, 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 can be improved. n is₁≈n₂≈n₃≈n₄≈n₅≈n₆The situation applies to polydisperse structures, which can satisfy the requirements of different molecular weights, and n₁＝n₂＝n₃＝n₄＝n₅＝n₆The 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-(Z₁)_q1-R₀₁，-(Z₂)_q-(Z₁)_q1-R₀₁See 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, and in this case, contains a functional group R₀₁。

1.6.1. Functional group R₀₁Definition of (1)

R₀₁Is 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 ₀₁Selected 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 bio-related 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 stablyBonds, degradable covalent bonds, dynamic covalent bonds. 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 oxidation, 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 modified form of the reactive group refers to a form (still reactive group) in which a reactive group remains active after at least one process of oxidation, reduction, hydration, dehydration, electronic rearrangement, structural rearrangement, salt complexation and decomplexing, ionization, protonation, deprotonation, substitution, deprotection, and the like, or a non-active 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-; 177-.

R₀₁Including but not limited to groups A through H or variations thereof, groups R₀₁Is a reactive group or a variant 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 active ester groups of similar structure A16-A18 (e.g., 2-thione-3-thiazolidine formate groups (tetrahydrothiazole-2-thione-N-formate groups), 2-sulfoximine-3-carboxylate groups, 2-pyrrolidine-N-carboxylate groups, 2-thione-pyrrolidine-N-formate groups, succinimide-ester groups, p-nitrophenyl, 2-thioketone benzothiazole-N-formate, 1-oxo-3-thiooxoisoindoline-N-formate, 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), tetramethylpiperidinoxy group, dioxopiperidinoxy group (3, 5-dioxo-1-cyclohexylamine-N-oxy group), ammonium salt (amine salt), hydrazine group, disulfide/disulfide compound (such as linear O-dithiopyridine and the like, e.g., cyclic lipoic acid and the like), 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, enamine group, alkynylamine group, or the like, Protected hydroxy or mercapto (carbamate, monothiocarbamate, dithiocarbamate), protected amino (carbamate, monothiocarbamate, dithiocarbamate), and 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, thiohydral group, thiothiothiothiothiothiothioketal group, thiothiothioketal group, thioester group (e.g., D26), thioester group (e.g., D27), dithioester group (e.g., D18), thiohemiacetal group, monothiohydrate group, dithiohydrate 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 E13₃Including 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 dienyls (e.g., furyl), 1,2,4, 5-tetrazinyls, and the like;

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, and the like), a protected hydroxyl group, a siloxy group, a protected dihydroxy group, a trihydroxysilyl group, a protected trihydroxysilyl group, and the like;

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), -NH₂Is ammonium ion-NH₃ ⁺Amine salt-NH₂HCl 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 ⁺)NH₂Is a protonated form of the guanidino group. One functional group can belong to both subcategories simultaneously. The ortho-pyridyl disulfide in C13 is also a protected form of the sulfhydryl group. 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. Within this class are predominantly groups with similar reactivity (e.g., hydroxylamine, hydrazine), protected forms, salt forms, and the like, and in addition include readily leaving halogens, and the like. 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 groups, the most common structures of which have similarities in preparation methods, can be obtained by substitution reactions of the 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.

The 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-tetrazine group can carry out cycloaddition 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 represented by G32 by treatment with hydrazine 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 thereof, 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 necessary constituents of the initiator core for initiating ethylene oxide polymerization 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.

R₀₁Or does not react with the biologically relevant substance, and has special functions, including two types of functional groups, namely a targeting group and a fluorescent group, or substituted forms thereof. The substituted forms need to still have the corresponding special function and can be classified as corresponding targeting groups and fluorescent groups. Such R₀₁Including 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 further include, but are not limited to, the rhodamine derivatives described in the document { Progress in Chemistry,2010,22(10):1929-1939} and citations thereof.

In the present invention, - (Z)₁)_q1-R₀₁Functional groups as a whole. Wherein, for example, R₀₁Functional groups include, but are not limited to, groups described and exemplified in documents CN104877127A, CN104530413A, CN104530415A, CN104530417A, including, but not limited to, CN104530417A, and the like, which are active esters, amino groups, aldehyde groups, carboxyl groups, acid halides, acid anhydrides, cyano groups, alkynyl groups, hydroxyl groups, and the like[0423]～[0432]Section CN104877127A [0308]～[0328]For example, they are not described in detail herein.

1.6.3. Functional group R₀₁Structural classification of

Specifically, R₀₁Including 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:

or class B:

or class C:

or class D:

or class E:

or class F:

or class G:

class Ga:

or class Gb:

or class H:

or class I:

or class J:

and the like.

Wherein Q is₅、M₅And M₅The rings are consistent with the above definitions and are not described further herein.

Wherein, Y₁Is a leaving group attached to a sulfonyl, sulfinyl, oxysulfonyl or oxysulfinyl group. Y is ₁There is no particular limitation. Y is₁Preferably having C_1-10Hydrocarbyl or fluoro C_1-10A hydrocarbyl group. Y is₁More preferably having C_1-10Alkyl radical, C_1-10Alkenyl, 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 Y₁Can 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 is₁More 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, W₃Is a leaving group including, but not limited to, F, Cl, Br, I, PhS-, preferably Br or Cl.

Wherein, W₂Is F, Cl, Br or I, preferably I.

Wherein the content of the first and second substances,

each of which is a cyclic structure containing a nitrogen atom, a nitrogen onium ion, a double bond, an azo, a triple bond, a disulfide bond, an anhydride, an imide, a diene in the ring backbone, including but not limited to a carbocycle, a heterocycle, a benzoheterocycle, a substituted carbocycle, a substituted heterocycle, or a substituted benzoheterocycle, and the like.

Wherein M is a carbon or heteroatom in the ring, including but not limited to carbon, nitrogen, phosphorus, silicon.

Wherein M is₈Is a carbon atom or a heteroatom located on the ring. M₈Preferably a carbon atom, a nitrogen atom, a phosphorus atom or a silicon atom. M₈The 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. M₈May be a carbon atom or a hetero atom on a 4-to 50-membered ring, preferably on a 4-to 32-membered ringA carbon atom, a nitrogen atom, a phosphorus atom or a silicon atom, more preferably a carbon atom, a nitrogen atom, a phosphorus atom or a silicon atom on a 5-32-membered ring, still more preferably a carbon atom, a nitrogen atom, a phosphorus atom or a silicon atom on 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 is₂₂Is 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. M₂₂The number of ring-forming atoms of the ring is 4, 5, 6, 7 or 8, preferably 4, 5 or 6.

Wherein PG₈Is 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 (Picture experts group) ₈May be a single trivalent end group such as

Taking D8 as an example, corresponding to

PG₈It may also be two or three separate end groups, and D8 corresponds to

H5 corresponds to

Wherein R is₂Is 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.

R₂Can be selected from hydrogen atoms, R₂₁Or R₃Any one atom or group.

Wherein R is₂₁Is a divalent linking group and participates in ring formation.

R₂₁The number of carbon atoms of (A) is not particularly limited, but preferably 1 to 20, more preferably 1 to10。

R₂₁The 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.

R₂₁May or may not contain heteroatoms.

R₂₁Is selected from C_1-20Alkylene, divalent C_1-20Heterohydrocarbyl, substituted C_1-20Alkylene, substituted divalent C_1-20Any divalent linking group or any combination of two or three of the divalent linking groups in the heterohydrocarbyl group. Wherein, the substituent atom or substituent is not particularly limited, including but not limited to any substituent atom or any substituent group of the term moiety, selected from any one of halogen atom, hydrocarbyl substituent group, heteroatom-containing substituent group.

R₂₁Preferably C_1-20Open-chain alkylene, C_1-20Alkenyl radical, C_1-20Cycloalkylene radical, C_1-20Cycloalkylene, arylene, divalent C_1-20Aliphatic heteroalkyl, divalent C_1-20Lipoheteroalkenyl, divalent heteroaryl, divalent heteroarylalkyl, substituted alkylene, substituted C_1-20Open alkenylene, substituted C_1-20Cycloalkylene, substituted C_1-20Cycloalkylene radical, substituted arylene radical, substituted divalent C radical_1-20Lipoheteroalkyl, substituted divalent C_1-20Any 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.

R₂₁More preferably C_1-10Open-chain alkylene, C_1-10Alkenyl radical, C_3-10Cycloalkylene radical, C_1-10Cycloalkylene, arylene, divalent C_1-10Aliphatic heteroalkyl, divalent C_1-10Lipoheteroalkenyl, divalent heteroaryl, divalent heteroarylalkyl, substituted aryleneAlkyl, substituted C_1-10Open alkenylene, substituted C_1-10Cycloalkylene, substituted C_1-10Cycloalkylene radical, substituted arylene radical, substituted aralkylene radical, substituted divalent C_1-10Lipoheteroalkyl, substituted divalent C _1-10Any one of divalent linking groups of lipoheteroalkenyl, substituted divalent heteroaryl, substituted divalent heteroarylalkyl, a combination of any two, or a combination of any three.

Specifically, R₂₁Selected from the group consisting of methylene, ethylene, propylene, butylene, pentylene, hexylene, heptylene, octylene, nonylene, decylene, 1, 2-phenylene, benzylene, C_1-20Oxaalkylene, C_1-20Thiaalkylene group, C_1-20Any 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 group substituent and a heteroatom-containing substituent, and is preferably a halogen atom, an alkoxy group or a nitro group.

R₂₁More preferred are 1, 2-ethylene group and 1, 3-propylene group.

Wherein R is₃Are terminal groups to which an oxygen or sulfur group is attached.

R₃The number of carbon atoms of (A) is not particularly limited, but is preferably 1 to 20, more preferably 1 to 10.

R₃The 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.

R₃May or may not contain heteroatoms.

R₃Is selected from C_1-20Hydrocarbyl radical, C_1-20Heterohydrocarbyl radical, C_1-20Substituted hydrocarbyl radical, C_1-20Any of substituted heterohydrocarbyl groups. For substitution of R₃The heteroatom or substituent of (a) is not particularly limited and includes, but is not limited to, any heteroatom or substituent of the term moietyAny substituent is preferably selected from a halogen atom, a hydrocarbon group, and a heteroatom-containing substituent.

R₃Preferably C_1-20Alkyl radical, C_3-20Alkylene, aryl, C_1-20Aliphatic heterocarbyl, heteroaryl, heteroaromatic hydrocarbyl, substituted C_1-20Alkyl, substituted C_3-20Alkylene, substituted aryl, substituted aralkyl, substituted C_1-20Any 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.

R₃Preferably C_1-20Straight chain alkyl, C_1-20Branched alkyl radical, C_3-20Cycloalkyl, aryl, aralkyl, C_1-20Aliphatic heterocarbyl, heteroaryl, heteroaromatic hydrocarbyl, substituted C_1-20Straight chain alkyl, substituted C_1-20Branched alkyl, substituted C_3-20Cycloalkyl, substituted aryl, substituted arylalkyl, substituted C_1-20Any 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, preferably halogen atom, alkoxy, alkyl, aryl or nitro.

R₃More preferably C_1-10Straight chain alkyl, C_1-10Branched alkyl radical, C_3-10Cycloalkyl, aryl, aralkyl, C_1-20Aliphatic heterocarbyl, heteroaryl, heteroaromatic hydrocarbyl, substituted C_1-10Straight chain alkyl, substituted C_1-10Branched alkyl, substituted C_3-10Cycloalkyl, substituted aryl, substituted arylalkyl, substituted C_1-10Any 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, R₃Selected 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.

R₃Most preferred is methyl, ethyl or benzyl.

Wherein R is₄Is- (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, R₄Selected from any one of halogen atoms. Fluorine atoms are preferred.

When taken as a substituent, R₄The 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, R₄The 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.

When taken as a substituent, R₄May or may not contain heteroatoms.

R₄Selected from hydrogen atoms, halogen atoms, C_1-20Hydrocarbyl radical, C_1-20Heterohydrocarbyl, substituted C_1-20Hydrocarbyl or substituted heterohydrocarbyl. Wherein R is₄The 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 group.

R₄More preferably a hydrogen atom, a halogen atom, C_1-20Alkyl radical, C_1-20Unsaturated aliphatic, aryl, C _1-20Heterohydrocarbyl radical, C_1-20Hydrocarbyloxyacyl group, C_1-20Hydrocarbyl thioacyl, C_1-20Any atom or group of a hydrocarbylaminoacyl group, or a substituted version of any group. Wherein R is₄The acyl group in (1) is not particularly limited, including but not limited to any acyl type of the term moiety. R₄The acyl group in (1) is more preferably a carbonyl group or a thiocarbonyl group.

R₄More preferably a hydrogen atom, a halogen atom, C_1-20Alkyl radical, C_1-20Alkenyl, aryl, arylalkyl, C_1-20Aliphatic heterocarbyl, heteroaryl, heteroaromatic hydrocarbyl, C_1-20Alkoxyacyl, aryloxyacyl, C_1-20Alkylthio acyl, arylthio acyl, C_1-20Any one of an alkylaminoacyl group, an arylaminoacyl group, or a substituted version of any one of the groups.

R₄More preferably a hydrogen atom, a halogen atom, C_1-20Alkyl radical, C_1-20Alkenyl, aryl, arylalkyl, C_1-20Aliphatic heterocarbyl, heteroaryl, heteroaromatic hydrocarbyl, C_1-20Alkoxycarbonyl, aryloxycarbonyl, C_1-20Alkylthio carbonyl, arylthio carbonyl, C_1-20Alkylaminocarbonyl, arylaminocarbonyl, C_1-20Alkoxythiocarbonyl, aryloxylthiocarbonyl, C_1-20Alkylthio thiocarbonyl, arylthio thiocarbonyl, C_1-20Any one atom or group of an alkylaminothiocarbonyl group, arylaminothiocarbonyl group, or a substituted version of any one group.

Specifically, R₄Selected 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, nonadecyl group, eicosyl group, allyl group, propenyl group, vinyl group, phenyl group, methylphenyl group, butylphenyl group, benzyl group, methoxy groupCarbonyl, ethoxycarbonyl, phenoxycarbonyl, benzyloxycarbonyl, methylthiocarbonyl, ethylthiocarbonyl, phenylthiocarbonyl, benzylthiocarbonyl, ethylaminocarbonyl, benzylaminocarbonyl, methoxythiocarbonyl, ethoxythiocarbonyl, phenoxythiocarbonyl, benzyloxythiocarbonyl, methylthiothiocarbonyl, ethylthiothiocarbonyl, phenylthiothiocarbonyl, benzylthiothiocarbonyl, ethylaminothiocarbonyl, benzylaminothiocarbonyl, substituted C_1-20Alkyl, substituted C_1-20Alkenyl, substituted aryl, substituted arylalkyl, substituted C_1-20Aliphatic heterocarbyl, substituted heteroaryl, substituted heteroarylalkyl, substituted C_1-20Alkoxycarbonyl, substituted aryloxycarbonyl, substituted C _1-20Alkylthio carbonyl, substituted arylthio carbonyl, substituted C_1-20Alkylaminocarbonyl, substituted arylaminocarbonyl, substituted C_1-20Alkoxythiocarbonyl, substituted aryloxythiocarbonyl, substituted C_1-20Alkylthio thiocarbonyl, substituted arylthio thiocarbonyl, substituted C_1-20An 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 fluorine atom, a chlorine atom, a bromine atom, an iodine atom, an alkenyl group or a nitro group.

R₄More 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, benzylthiocarbonylthiocarbonyl group Radical, ethylaminothiocarbonyl, benzylaminothiocarbonyl, C_1-10Halogenated hydrocarbon group, halogenated phenyl, halogenated benzyl, nitro phenyl and any kind of atom or group, or any kind of substituted form of group.

R₄Preferably 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.

R₄Most preferably a hydrogen atom, a methyl group or a benzyl group.

Wherein R is₈、R₉、R₁₀、R₁₁、R₁₂Each independently is a hydrogen atom, a substituent atom or a substituent on a double bond (-C-), and R is in the same molecule₈、R₉、R₁₀、R₁₁、R₁₂May be the same as or different from each other.

When it is a substituted atom, R₈、R₉、R₁₀、R₁₁、R₁₂Each 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, R₈、R₉、R₁₀、R₁₁、R₁₂The number of carbon atoms of (a) is not particularly limited. R₈、R₉、R₁₀、R₁₁、R₁₂The 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, R₈、R₉、R₁₀、R₁₁、R₁₂The 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-mentioned aliphatic ring, aromatic ring, sugar ring, and condensed ring are preferable.

When it is a substituent, R₈、R₉、R₁₀、R₁₁、R₁₂Each independently may or may not contain a heteroatomAn atom.

R₈、R₉、R₁₀、R₁₁、R₁₂Each independently selected from a hydrogen atom, a halogen atom, C_1-20Hydrocarbyl radical, C_1-20Heterohydrocarbyl, substituted C_1-20Hydrocarbyl or substituted heterohydrocarbyl. Wherein R is₈The 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 group.

R₈、R₉、R₁₀、R₁₁、R₁₂Each independently more preferably a hydrogen atom, a halogen atom, C_1-20Alkyl radical, C_1-20Unsaturated aliphatic, aryl, C_1-20Heterohydrocarbyl radical, C_1-20Hydrocarbyloxyacyl group, C_1-20Hydrocarbyl thioacyl, C_1-20Any atom or group of a hydrocarbylaminoacyl group, or a substituted version of any group. Wherein R is₈The acyl group in (1) is not particularly limited, including but not limited to any acyl type of the term moiety.

R₈、R₉、R₁₀、R₁₁、R₁₂Each independently more preferably a hydrogen atom, a halogen atom, C_1-20Alkyl radical, C_1-20Alkenyl, aryl, arylalkyl, C_1-20Aliphatic heterocarbyl, heteroaryl, heteroaromatic hydrocarbyl, C_1-20Alkoxyacyl, aryloxyacyl, C_1-20Alkylthio acyl, arylthio acyl, C_1-20Any one of an alkylaminoacyl group, an arylaminoacyl group, or a substituted version of any one of the groups. 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

R₈、R₉、R₁₀、R₁₁、R₁₂Each independently more preferably a hydrogen atom, a halogen atom, C_1-20Alkyl radical, C_1-20Alkenyl, aryl, arylalkyl, C_1-20Aliphatic heterohydrocarbyl, heteroaryl, heteroaromatic hydrocarbyl,C_1-20Alkoxycarbonyl, aryloxycarbonyl, C_1-20Alkylthio carbonyl, arylthio carbonyl, C_1-20Alkylaminocarbonyl, arylaminocarbonyl, C_1-20Alkoxythiocarbonyl, aryloxylthiocarbonyl, C_1-20Alkylthio thiocarbonyl, arylthio thiocarbonyl, C_1-20Any one atom or group of an alkylaminothiocarbonyl group, arylaminothiocarbonyl group, or a substituted version of any one group. R₈The acyl group in (1) is more preferably a carbonyl group or a thiocarbonyl group. 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 fluorine atom, a chlorine atom, a bromine atom, an iodine atom, an alkenyl group or a nitro group.

Specifically, R₈、R₉、R₁₀、R₁₁、R₁₂Each independently 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, a 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 methylthiocarbonyl group, a benzyloxycarbonyl group, a methylthiocarbonyl group, an ethylthiocarbonyl group, Benzylthiothiocarbonyl, ethylaminothiocarbonyl, benzylaminothiocarbonyl, substituted C _1-20Alkyl, substituted C_1-20Alkenyl, substituted aryl, substituted arylalkyl, substituted C_1-20Aliphatic heterocarbyl, substituted heteroaryl, substituted heteroarylalkyl, substituted C_1-20Alkoxycarbonyl, substituted aryloxycarbonyl, substituted C_1-20Alkylthio carbonyl, substituted arylthio carbonyl, substituted C_1-20Alkylaminocarbonyl, substituted arylaminocarbonyl, substituted C_1-20Alkoxythiocarbonyl, substituted aryloxythiocarbonyl, substituted C_1-20Alkylthio thiocarbonyl, substituted arylthio thiocarbonyl, substituted C_1-20An 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.

R₈、R₉、R₁₀、R₁₁、R₁₂Further 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 group, methoxythiocarbonyl group, ethoxythiocarbonyl group, phenoxythiocarbonyl group, benzyloxythiocarbonyl group, methylthiothiocarbonyl group, ethylthiocarbonyl group, phenylthiocarbonyl group, benzylthiocarbonyl group, ethylaminothiocarbonyl group, benzylaminothiocarbonyl group, C thiocarbonyl group, C _1-10Halogenated hydrocarbon group, halogenated phenyl, halogenated benzyl, nitro phenyl and any kind of atom or group, or any kind of substituted form of group. 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 fluorine atom, a chlorine atom, a bromine atom, an iodine atom, an alkenyl group or a nitro group.

R₈、R₉、R₁₀、R₁₁、R₁₂Each independently is more preferably a hydrogen atom, a fluorine atom or a methyl group.

In class E3, R₈Most preferred is methyl.

Wherein R is₂₄To end groups of disulfide bondsPreferably C_1-20Alkyl, aryl, heteroaryl, and the like, such as ortho-pyridyl.

Wherein R is₂₇For the substituent attached to the azo, a phenyl group, a substituted phenyl group or a hybridized phenyl group is preferred.

Wherein R is₃₀Is a hydrocarbon radical, preferably C_1-20Alkyl, benzyl, phenyl ring hydrogen atoms by C_1-20Hydrocarbyl-substituted benzyl.

Wherein M is₁₉、M₂₀、M₂₁Each independently an oxygen atom or a sulfur atom, and may be the same as or different from each other in the same molecule.

Wherein, X₁₁To attach a carbonyl or thiocarbonyl end group, preferably C_1-20Alkyl groups, more preferably methyl, ethyl, isopropyl, tert-butyl.

Wherein, X₁₂Terminal groups to which carbonate or thiocarbonate groups are attached, selected from hydrocarbon groups (which may or may not include a benzene ring), preferably C _1-20Hydrocarbyl, more preferably C_1-20Alkyl, phenylhydrocarbyl or hydrocarbyl substituted phenyl.

Wherein, X₁₃Is a terminal group for attachment of a thio group, selected from a mercapto-protecting group or the group LG₂。

When it is a mercapto-protecting group, X₁₃Is selected from PG₂Thiol protecting groups in the listed groups.

Wherein LG is₂The number of carbon atoms of (a) is not particularly limited. LG (Ligno-lead-acid)₂The number of carbon atoms of (A) is preferably 1 to 20, more preferably 1 to 10.

LG₂The 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.

LG₂May or may not contain heteroatoms.

LG₂Is selected from C_1-20Hydrocarbyl radical, C_1-20Heterohydrocarbyl, substituted C_1-20Any one of a hydrocarbon group and a substituted heterohydrocarbon group. Wherein LG is₂The 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.

LG₂More preferably C_1-20Alkyl radical, C_1-20Unsaturated aliphatic, aryl, C_1-20Heterohydrocarbyl radical, C_1-20Alkylthio radical, C_1-20Aliphatic heterocarbylthio, arylthio, C _1-20Fatty hydrocarbyl acyl radical, C_1-20Lipoheteroalkylacyl, arylacyl, heteroarylacyl, C_1-20Hydrocarbyloxyacyl group, C_1-20Hydrocarbyl thioacyl, C_1-20Hydrocarbyl aminoacyl radical, C_1-20Heterohydrocarbyloxyacyl group, C_1-20Heterocarbylthioacyl radical, C_1-20Any one group or substituted version of any one group of a heterocarbylaminoacyl group. Wherein LG is₂The acyl group in (1) is not particularly limited, including but not limited to any acyl type of the term moiety. By way of example, LG₂The acyl group in (1) may be selected from a carbonyl group, a sulfonyl group, a sulfinyl group, a phosphoryl group, a hypophosphoryl group, a nitroxyl group, a nitrosyl group, a thiocarbonyl group, an imidoyl group, a thiophosphoryl group, a dithiophosphoryl group, a trithiophosphoryl group, a thiophosphorous group, a dithiophosphoryl group, a thiophosphoryl group, a dithiophosphoryl group, a thiophosphoryl 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)₂The acyl group in (1) is more preferably a carbonyl group, thiocarbonyl group or sulfonyl group.

LG₂More preferably C_1-20Alkyl, aryl, aralkyl, C_1-20Heteroalkyl, heteroaryl, heteroaralkyl, C_1-20Alkylthio, arylthio, aralkylthio, C _1-20Heteroalkylthio, heteroarylthio, heteroaralkylthio, C_1-20Alkylcarbonyl, arylcarbonyl, aralkylcarbonyl, C_1-20Heteroalkylcarbonyl, heteroarylcarbonyl, heteroaralkylcarbonyl, C_1-20Alkoxycarbonyl, aryloxycarbonyl, aralkyloxycarbonyl, C_1-20Alkylthio-carbonyl, arylthio-carbonyl, aralkylthiocarbonyl, C_1-20Alkylaminocarbonyl, arylaminocarbonyl, aralkylaminocarbonyl, C_1-20Heteroalkyloxycarbonyl, heteroaryloxycarbonyl, heteroaralkyloxycarbonyl, C_1-20Heteroalkylthio-carbonyl, heteroarylthio-carbonyl, heteroaralkylthio-carbonyl, C_1-20Heteroalkylaminocarbonyl, heteroarylaminocarbonyl, heteroarylalkylaminocarbonyl, C_1-20Alkylthio, arylthio, aralkylthiocarbonyl, C_1-20Heteroalkylthiocarbonyl, heteroarylthiocarbonyl, heteroarylalkylthiocarbonyl, C_1-20Alkoxythiocarbonyl, aryloxylthiocarbonyl, aralkyloxythiocarbonyl, C_1-20Alkylthio thiocarbonyl, arylthio thiocarbonyl, aralkylthio thiocarbonyl, C_1-20Alkylaminothiocarbonyl, arylaminothiocarbonyl, aralkylaminothiocarbonyl, C_1-20Heteroalkyloxythiocarbonyl, heteroaryloxythiocarbonyl, heteroarylalkoxythiocarbonyl, C _1-20Heteroalkylthio thiocarbonyl, heteroarylthio thiocarbonyl, heteroarylalkylthio thiocarbonyl, C_1-20A heteroalkylaminothiocarbonyl group, a heteroarylaminothiocarbonyl group, or a substituted version of any group.

LG₂More preferably C_1-20Alkyl, aryl, aralkyl, C_1-20Heteroalkyl, heteroaryl, heteroaralkyl, C_1-20Alkylthio, arylthio, aralkylthio, C_1-20Any one of the groups or substituted versions of any one of the groups heteroarylthio, heteroaralkylthio.

Specifically, LG₂Selected from the group consisting of, 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, ethyl acyl, benzoyl, methoxyacyl, ethoxy acyl, tert-butyloxyacyl, phenoxy acyl, benzyloxy acyl, methylthioacyl, ethylthioacyl, tert-butylthioacyl, phenylthioacyl, benzylthioacyl, 2-pyridylcarbonyl, methylaminoacyl, ethylaminoacyl, tert-butylaminoacyl, benzylaminoacyl and the like, or a substituted form of any group. 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 heteroatom-containing substituent, and is preferably a fluorine atom, a chlorine atom, a bromine atom, an iodine atom or a nitro group.

LG₂More preferred 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, benzyl, methylbenzyl, nitrobenzyl, tert-butylthio, benzylthio, 2-pyridylthio, acetyl, benzoyl, methoxycarbonyl, ethoxycarbonyl, tert-butyloxycarbonyl, phenoxycarbonyl, benzyloxycarbonyl, methylthiocarbonyl, ethylthiocarbonyl, tert-butylthiocarbonyl, phenylthiocarbonyl, benzylthiocarbonyl, 2-pyridylcarbonyl, methylaminocarbonyl, ethylaminocarbonyl, tert-butylaminocarbonyl, benzylaminocarbonyl, ethylthiocarbonyl, tert-butylaminocarbonyl, benzylthiocarbonyl, tert-butylaminocarbonyl, tert-propyl, Methylthiocarbonyl, ethoxythiocarbonyl, tert-butyloxythiocarbonyl, phenoxythiocarbonyl, benzyloxythiocarbonyl, methylthiothiocarbonyl, ethylthiothiocarbonyl, tert-butylthiothiocarbonyl, phenylthiothiocarbonyl, benzylthiocarbonyl, methylaminothiocarbonyl, ethylaminothiocarbonyl, tert-butylaminothiocarbonyl, benzylaminothiocarbonyl, C _1-10Any one of halogenated hydrocarbon group, trifluoroacetyl group, halogenated phenyl group, halogenated benzyl group, nitrophenyl group and the like, or any one of the groups may be substitutedForm (a). Among them, the substituent atom or the substituent is preferably a fluorine atom, an alkoxy group or a nitro group.

LG₂More 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-butylthiothiocarbonyl, phenylthiothiocarbonyl, benzylthiothiocarbonyl, trifluoroacetyl and the like.

LG₂More preferably, it is any of tert-butyl, trityl, phenyl, benzyl, methylbenzyl, tert-butylthio, benzylthio, 2-pyridylthio and the like.

LG₂Most preferred is methyl, ethyl, allyl or benzyl.

Wherein, X₆Is a terminal group attached to the oxygen atom of the ester group and is selected from a hydroxyl protecting group or the group LG₄. Wherein LG is₄The 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 structures are used, the same structure may be used, or a combination of two or more different structures may be used.

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 _1-20Haloalkyl, C_1-20Alkyl radical, C_2-20Alkenyl radical, C_3-20Open-chain alkenyl, C_3-20Cycloalkyl, aryl, arylalkyl, C_1-20Heteroalkyl, heteroaryl, heteroaralkyl, C_1-20Alkoxy, aryloxy, aralkyloxy, C_1-20Heteroalkyloxy, heteroaryloxy, heteroarylhydrocarbyloxy, C_1-20Alkylthio, arylthio, aralkylthio, C_1-20Any 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 any substituted heteroatom or any substituent of the term moiety, selected from any of halogen atoms, hydrocarbyl substituents, heteroatom-containing substituents.

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, a thioester group-containing substituent at the terminal, an amide bond-containing substituent at the terminal, C_1-20Haloalkyl, C_2-20Alkenyl radical, C_3-20Open-chain alkenyl, C_3-20Cycloalkyl, aryl, arylalkyl, C_1-20Heteroalkyl, heteroaryl, heteroaralkyl, C_1-20Alkoxy, aryloxy, aralkyloxy, C_1-20Heteroalkyloxy, heteroaryloxy, heteroarylhydrocarbyloxy, C _1-20Alkylthio, arylthio, aralkylthio, C_1-20Any one atom or group, or substituted version of any one group, of heteroalkylthio, heteroarylthio, heteroarylalkylthio, and the like. Wherein the acyl group is not particularly limited, including but not limited to any acyl type of the term moiety. By way of example, in QThe acyl group of (a) may be selected from carbonyl, sulfonyl, sulfinyl, phosphoryl, phosphorylidene, nitroxyl, nitrosyl, thiocarbonyl, imidoyl, thiophosphoryl, dithiophosphoryl, trithiophosphoryl, thiophosphoridene, dithiophosphorylidene, thiohypophosphoryl, thiophosphoryl, dithiophosphoryl, thiophosphoryl 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_1-20Carbonyl group, C_1-20Alkylthio carbonyl of C_1-20Sulfonyl radical, C_1-20Alkyloxycarbonyl, C_1-20Alkylthio carbonyl group, C_1-20Alkylaminocarbonyl radical, C_1-20Alkyloxythiocarbonyl radical, C _1-20Alkylthio thiocarbonyl radical, C_1-20Alkylamino thiocarbonyl radical, C_1-20Alkyloxysulfonyl, C_1-20Alkyloxysulfinyl, arylthiocarbonyl, aryloxycarbonyl, arylthiocarbonyl, arylaminocarbonyl, aryloxysulfonyl, aryloxysulfinyl, aralkylthiocarbonyl, aralkyloxycarbonyl, aralkylthiocarbonyl, aralkylaminocarbonyl, aralkyloxythiocarbonyl, aralkylthiothiocarbonyl, aralkylaminothiocarbonyl, aralkyloxysulfonyl, aralkyloxysulfinyl, C_1-20Alkyl radical, C_2-20Alkenyl radical, C_3-20Open-chain alkenyl, C_3-20Cycloalkyl, aryl, arylalkyl, C_1-20Heteroalkyl, heteroaryl, heteroaralkyl, C_1-20Alkoxy, aryloxy, aralkyloxy, C_1-20Heteroalkyloxy, heteroaryloxy, heteroarylhydrocarbyloxy, C_1-20Alkylthio, arylthio, aralkylthio, C_1-20Heteroalkylthio, heteroarylthio, heteroarylalkylthio, C_1-20Any atom or group of haloalkyl, etc., or substituted form of any groupFormula (II) is shown.

Q is more preferably a hydrogen atom, a halogen atom, a nitro group-containing substituent, or C _1-10Carbonyl group, C_1-10Alkylthio carbonyl of C_1-10Sulfonyl radical, C_1-10Alkyloxycarbonyl, C_1-10Alkylthio carbonyl group, C_1-10Alkylaminocarbonyl radical, C_1-10Alkyloxythiocarbonyl radical, C_1-10Alkylthio thiocarbonyl radical, C_1-10Alkylamino thiocarbonyl radical, C_1-10Alkyloxysulfonyl, C_1-10Alkyloxysulfinyl, arylthiocarbonyl, aryloxycarbonyl, arylthiocarbonyl, arylaminocarbonyl, aryloxysulfonyl, aryloxysulfinyl, aralkylthiocarbonyl, aralkyloxycarbonyl, aralkylthiocarbonyl, aralkylaminocarbonyl, aralkyloxythiocarbonyl, aralkylthiothiocarbonyl, aralkylaminothiocarbonyl, aralkyloxysulfonyl, aralkyloxysulfinyl, C_1-20Alkyl radical, C_2-10Alkenyl radical, C_3-10Open-chain alkenyl, C_3-10Cycloalkyl, aryl, arylalkyl, C_1-10Heteroalkyl, heteroaryl, heteroaralkyl, C_1-10Alkoxy, aryloxy, aralkyloxy, C_1-10Heteroalkyloxy, heteroaryloxy, heteroarylhydrocarbyloxy, C_1-10Alkylthio, arylthio, aralkylthio, C_1-10Heteroalkylthio, heteroarylthio, heteroarylalkylthio, C _1-10Haloalkyl, and the like, or substituted versions of either 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 groupCarbonyl, ethoxythiocarbonyl, tert-butyloxythiocarbonyl, phenoxythiocarbonyl, benzyloxythiocarbonyl, methylthiothiocarbonyl, ethylthiocarbonyl, tert-butylthiocarbonyl, phenylthiocarbonyl, benzylthiocarbonyl, ethylaminoacyl, tert-butylaminothiocarbonyl, 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, n-butylphenyl, p-butylphenyl, n-dodecyl, n-eicosyl, vinyl, propenyl, allyl, propynyl, Phenoxy, benzyloxy, methylthio, ethylthio, phenylthio, benzylthio, C _1-20Haloalkyl, 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, 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, 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 cyclop, Trifluoromethyl, 2,2, 2-trifluoroethyl, and the like, or a substituted version of any. 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 is₃Is an H atom or a group that contributes to the induction, conjugation effect of unsaturated bond electrons;

Q₃selected 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.

Q₃May 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.

Q₃The 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.

Q₃Can 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 ₃The 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.

Q₃More preferably a hydrogen atom, a halogen atom, C_1-20Alkyl radical, C_2-20Alkenyl radical, C_3-20Open-chain alkenyl, C_3-20Cycloalkyl, aryl, arylalkyl,C_1-20Heteroalkyl, heteroaryl, heteroaralkyl, C_1-20Alkoxy, aryloxy, aralkyloxy, C_1-20Heteroalkyloxy, heteroaryloxy, heteroarylhydrocarbyloxy, C_1-20Heteroalkylthio, heteroarylthio, heteroarylalkylthio, C_1-20Haloalkyl, and the like, or substituted versions of either group.

Q₃More preferably a hydrogen atom, a halogen atom, C_1-10Haloalkyl, C_1-10Alkyl radical, C_2-10Alkenyl radical, C_3-10Open-chain alkenyl, C_3-10Cycloalkyl, aryl, arylalkyl, C_1-10Heteroalkyl, heteroaryl, heteroaralkyl, C_1-10Alkoxy, aryloxy, aralkyloxy, C_1-10Any atom or group, or substituted version of any group, of heteroalkyloxy, heteroaryloxy, heteroarylhydrocarbyloxy, and the like.

Specifically, Q₃Can be selected from 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, 2-ethylhexyl group, nonyl group, decyl group, undecyl group, dodecyl group, tridecyl group, tetradecyl group, pentadecyl group, hexadecyl group, heptadecyl group, octadecyl group, nonadecyl group, eicosyl group, vinyl group, propenyl group, allyl group, propynyl group, propargyl group, cyclopropyl group, cyclopropenyl group, phenyl group, benzyl group, butylphenyl group, p-methylphenyl group, nitrophenyl group, p-methoxyphenyl group, azaphenyl group, methoxy group, ethoxy group, phenoxy group, benzyloxy group, methylthio group, ethylthio group, phenylthio group, benzylthio group, C _1-20Haloalkyl, 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, 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.

Q₃Preferably a hydrogen atom, a fluorine atom, a chlorine atom, a bromine atom, an iodine atom, a methyl groupAn atom or group selected from the group consisting of ethyl, n-propyl, isopropyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, vinyl, propenyl, allyl, propynyl, propargyl, cyclopropyl, cyclopropenyl, phenyl, benzyl, butylphenyl, p-methylphenyl, p-nitrophenyl, o-nitrophenyl, p-methoxyphenyl, azaphenyl, methoxy, ethoxy, phenoxy, benzyloxy, methylthio, ethylthio, phenylthio, benzylthio, trifluoromethyl, 2,2, 2-trifluoroethyl, and a substituted form of any of these 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.

Q₃More preferably any 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 diaza-phenyl group or a substituted form thereof, a triaza-phenyl group or a substituted form thereof, and the like.

Q₃More preferably a hydrogen atom, methyl group, phenyl group, pyridyl group, diazophenyl group, triazophenyl group.

Q₃More preferably a hydrogen atom, a methyl group, a phenyl group or a pyridyl group.

Q₃Most preferably a hydrogen atom, a phenyl group or a pyridyl group.

Wherein Q is₆Is a hydrogen atom or a methyl group. Q₇Is hydrogen atom, methyl, phenyl or substituted phenyl. Such as p-methoxyphenyl. In the same molecule, Q₆And Q₇May be the same or different.

Wherein Q is₈The 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 is₈May 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 is₁₁Is a substituent on the nitrogen atom of tetrazole, preferably phenyl, substituted phenyl or aza phenyl.

Wherein PG₂Is a thiol protecting group, the protected thiol group being denoted as SPG ₂。

Wherein PG₃Being an alkynyl protecting group, the protected alkynyl group is represented by C ≡ CPG₃。

Wherein PG₄Is a hydroxy protecting group, the protected hydroxy group being represented by OPG₄。

Wherein PG₅Is an amino protecting group, the protected amino group being represented by NPG₅。

PG₂、SPG₂、PG₃、PG₄、OPG、PG₅、NPG₅Including 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 SPG₂Preferred are thioether, disulfide, silyl sulfide, thioester, and the like. In particular, SPG₂Any one of tert-butyl sulfide, trityl sulfide, substituted trityl sulfide, tert-butyl dimethyl silyl sulfide, triisopropyl silyl sulfide, benzyl sulfide, substituted benzyl sulfide, p-nitrobenzyl sulfide, o-nitrobenzyl sulfide, acetyl thioester, benzoyl thioester, trifluoroacetyl thioester, tert-butyl disulfide, substituted phenyl disulfide, 2-pyridine disulfide and the like is preferable.

The PG₃Preferred 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 PG₄It may be a protecting group for alcoholic hydroxyl group or phenolic hydroxyl group. OPG₄Preferred are ether, silyl ether, ester, carbonate, sulfonate, and the like. In particular, OPG₄Preferably methyl ether, 1-ethoxyethyl ether, tert-butyl ether, allyl ether, benzyl ether,P-methoxybenzyl ether, o-nitrobenzyl ether, p-nitrobenzyl ether, 2-trifluoromethylbenzyl ether, methoxymethyl ether, 2-methoxyethoxymethyl ether, benzyloxymethyl ether, p-methoxybenzyloxymethyl ether, methylthiomethyl ether, tetrahydropyranyl ether, trimethylsilyl ether, triethylsilyl ether, triisopropylsilyl ether, tert-butyldimethylsilyl ether, acetate, chloroacetate, trifluoroacetate, carbonate, and the like. Among the ether protective 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 PG₅May be a protecting group for primary amine, secondary amine, hydrazine, etc. NPG ₅Preferred are structures such as carbamate, amide, imide, N-alkylamine, N-arylamine, imine, enamine, imidazole, pyrrole, indole, and the like. In particular, NPG₅Preferably, it is any of formamide, acetamide, trifluoroacetamide, tert-butyl carbamate, 2-iodoethyl carbamate, benzyl carbamate, 9-fluorenemethyl carbamate, 2-trimethylsilylethyl carbamate, 2-methylsulfonylethyl carbamate, 2- (p-toluenesulfonyl) ethyl carbamate, phthalimide, diphenylmethyleneamine, 1,3, 5-dioxoazacyclohexyl, methylamino, triphenylmethylamino, tert-butylamino, allylamino, benzylamino, 4-methoxybenzylamino, benzylimine, and the like. Wherein PG₆Is a bishydroxy protecting group, and PG₆And two oxygen atoms form an acetal structure of a five-membered ring or a six-membered ring. PG (Picture experts group)₆Selected from methylene or substituted methylene. The PG₆The 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-R₀₁Examples of (2)

By way of example, - (Z)₂)_q-(Z₁)_q1-R₀₁Including 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_A0A1、L₂、L₀(g＝1)、Z₁、Z₂Any divalent linking group, or a divalent linking group consisting of any divalent linking group and an adjacent hetero atom group, may also be present in any polyvalent group A₀、A₁G, or any divalent linking group formed by a polyvalent group and an adjacent group.

The hexa-arm polyethylene glycol derivative may be stable 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 located at L₀、G、(Z₂)_q-(Z₁)_q1Any one of the degradable groups; (2) but not for Z, at the junction of any of the above groups with an adjacent group₁-R₀₁The 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、L₀-G、G-Z₂、Z₂-Z₁Degradation of either attachment site can occur.

According to the number of degradable sites in the six-arm polyethylene glycol derivative and the difference of the positions of the degradable sites, the stability of the polymer and the releaseability of the modified drug are greatly influenced. When degradation can occur between the functional groups at the ends of the six polyethylene glycol chains and the polyethylene glycol chain, i.e., - (Z)₂)_q-(Z₁)_q1The position of-the drug molecule and the polyethylene glycol structure are separated, so that the drug isThe active site of the molecule is exposed to the greatest extent, and the drug molecule can approach the unmodified state to the greatest extent.

Several degradation modes are typical as follows: (a) when g is 0 or 1, when only in L₂When the connection position of the PEG-PEG; (b) when g is 0 or 1, when g is only in- (Z)₂)_q-(Z₁)_q1-a position comprising its linker with the adjacent group on the PEG side is degraded to a six-arm PEG structure with an independent functional residue; (c) when g is 1, and is only in L₀Position (including L)₀Internal, O-L₀Connection, L₀-G-attachment) to a six-arm PEG structure and a cluster of multiple functional groups attached via G; (d) when G is 1, and degradation occurs only in G, degradation occurs to hexa-arm PEG, independent functional group residues, or residues of multiple functional clusters.

The hexa-arm polyethylene glycol derivative allows 1 or more than 1 degradation modes to exist. 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 the STAG is not particularly limited, and the stable presence condition may be 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 any condition including light, heat, enzyme, redox, acidic, alkaline, and the like.

The type of STAG is not particularly limited and includes, but is 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 bonds, thioether bonds, urea bonds, thiourea bonds, carbamate groups, thiocarbamate groups, divalent silicon groups containing no active hydrogen, divalent linking groups containing a boron atom, secondary amino groups, tertiary amino groups, Carbonyl, thiocarbonyl, acylamino, thioamido, sulfamide, enamine, triazolyl, 4, 5-dihydroisoxazolyl, any divalent connecting group in skeleton of amino acid and its derivative, and stable divalent connecting group formed from any two or more than two 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 divalent 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, -CH₂O-、-OCH₂-、-CH₂CH₂O-、-OCH₂CH₂-、-OCH₂CH₂O-、-(CH₂)₃O-、-O(CH₂)₃-、-(CH₂)₃O-、-O(CH₂)₃-and the like. As examples of L₀An 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, L₀In one aspect, the composition may further comprise (L)₇O)_nj-、-(OL₇)_nj-、-(R₂₉O)_nj-、-(OR₂₉)_nj-、-(CH₂CH₂O)_nj-、-(OCH₂CH₂)_nj-and the like. Wherein L is₇、R₂₉The 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. But of the invention CORE₈(O-)₈Does not include the removal of-CH₂CH₂A heteroatom-containing repeating unit other than O-.

1.7.2. Degradable divalent linking 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 the degradable divalent linking group containing the aromatic ring, the aromatic ring and the degradable divalent linking group can be combined.

The type of DEGG is not particularly limited and includes, but is not limited to, compounds 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, thioketal groups, azaketal groups, azathioketal groups, azothioketal groups, imine bonds, hydrazone bonds, acylhydrazone bonds, oxime bonds, sulfoximine ether groups, semicarbazone bonds, thiosemicarbazone bonds, hydrazine groups, hydrazide groups, thiocarbohydrazide groups, azocarbohydrazide groups, hydrazinocarbohydrazide groups, hydrazinoformate groups, carbazate groups, thiosemicarbazide groups, dithiocarbonate groups, dithioacetal groups, dithio, Hydrazinothiocarbamate groups, carbazolyl groups, thiocarbazohydrazine groups, azo groups, isoureido groups, isothioureido groups, allophanate groups, thioallophanate groups, guanidino groups, amidino groups, aminoguanidino groups, aminoamidino groups, imino groups, iminothioester groups, sulfonate groups, sulfinate groups, sulfonylhydrazide groups, sulfonylurea groups, maleimide groups, orthoester groups, phosphate groups, 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.

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, but not limited to, groups composed of a stable trivalent group containing a trivalent atomic nucleus structure and a degradable divalent linking group, groups composed of a trivalent aromatic ring and a degradable divalent linking group, combinations of a degradable trivalent ring structure and a divalent linking group that can exist stably, combinations of a degradable trivalent ring structure and a degradable divalent linking group, trivalent forms of any of the degradable divalent linking groups described above. 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, such as cyclic structures 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-containing, comb-like, dendritic, hyperbranched, and the like types. G may be degradable or may exist stably.

L₀The divalent linking group, which is used to link the PEG segment to the terminal branched structure G, may or may not be present. L is₀May be stable or degradable. L is₀May be selected from any of the foregoing STAG or DEGG.

The terminal branched groups G of the six-arm polyethylene glycol derivatives have the same structural type, such as a triple-branched structure, or a quadruple-branched structure, or a comb-like structure, or a tree-like structure, or a hyperbranched structure, or a cyclic structure. In the case of the same structure type, the structures allowing six PEG chain ends are 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 a comb-shaped or hyperbranched structure, the k at the end can 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 six-arm polyethylene glycol derivative G is 1, G is selected from the group including, but not limited to, any of the above groups having a valence of k +1(k is 2 to 250). 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, including but not limited to the above group G³Trivalent radical of (1), E_i(i＝1,2,3or 4)。L₀-G preferably comprises a structure selected from the group consisting of: e₀，

And the like; wherein, the asterisk marks in the structure, which indicates that the asterisk end points to the polyethylene glycol unit; said E₀Is any one of A₀And preferred forms thereof, any of A₁And preferred forms thereof(ii) a Said E₀May also be a trivalent skeleton structure of an amino acid or a derivative thereof; wherein the amino acid is_LIs of type or_D-type; the amino acid or the derivative thereof is derived from any one of the following: serine, threonine, cysteine, tyrosine, 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 ⁴A tetravalent group of (1); tetravalent G preferably contains the atom CM₄And unsaturated bond CB₄Ring structure CC₄Either tetravalent core structure, or contains two trivalent core structures. L is₀-G further preferably comprises any of the following structures:

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+1The 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 L₁₀Indirectly 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

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 there are twoOr more than two L₁₀When L is₁₀May be the same as or different from each other. L is₁₀The definitions of (a) and (b) are 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)_denrNONE, d) or DENR (U)_denr,L₁₀D) represents U_denrRepresents a polyvalent radical repeating unit, NONE represents a direct linkage of polyvalent repeating units, L₁₀Denotes a polyvalent repeating unit via a divalent linking group L₁₀Indirectly 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

And the like. Wherein ng represents the algebra of the 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

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

Is selected from

Wherein the asterisks in the structure indicate that the asterisk ends point towards 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 units of the polyvalent G constituting the comb-like composite structure include, but are not limited to, polyglycerin, polypentaerythritol, substituted propylene oxide, a group of substituted propylene oxide and carbon dioxide, acrylate and derivatives thereof, methacrylate and derivatives thereof, acetal structure-containing basic units (e.g., (1 → 6) β -D glucopyranoside), hydroxyl or thio group-containing amino acids and derivatives thereof, acidic amino acids and derivatives thereof, basic amino acids and derivatives thereof, 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 any form in which the hydroxyl groups other than the hydroxyl groups of the ether bond are protected, such as glycerol, trimethylolethane, trimethylolpropane.

Among them, the polyvalent G in 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 G⁵A pentavalent group in (1). Pentavalent G can include 1 pentavalent nuclear structure, 1 tetravalent nuclear structure and 1 trivalent nuclear structure, or 3 trivalent nuclear structures. L is₀-G preferably contains any one of the following structures:

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 triglutamic acid skeleton, a tripolyaspartic acid skeleton, a triglycerol skeleton, etc., such as

(ii) a A comb structure formed by indirectly combining 3 trivalent groups, such as three lysines combined by taking amino acids such as glycine, alanine and the like as spacers. Wherein the asterisks in the structure indicate that the asterisk ends point towards the polyethylene glycol unit.

For example, when the terminal reaction site k is 5, G is a hexavalent group, including but not limited to the above set G⁶The hexavalent group of (1). Hexavalent G may include 1 hexavalent nucleus structure, 1 pentavalent nucleus structure and 1 trivalent nucleus structure, 2 tetravalent nucleus structures, 1 tetravalent nucleus structure2 trivalent nuclear structures, or 4 trivalent nuclear structures. L is₀-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.),

and the like. Wherein the asterisks in the structure indicate that the asterisk ends point towards the polyethylene glycol unit.

2. Preparation method of six-arm polyethylene glycol derivative

2.1. A preparation method of a hexa-arm polyethylene glycol derivative relates to the following steps:

step one, adopting a hexahydroxy micromolecule containing six hydroxyl groups

The initiator system of (1); wherein, deprotonation of six exposed hydroxyl groups forms hexaoxide anions

Is stable under anionic polymerization conditions;

initiating ethylene oxide polymerization;

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, dimethylformamide or dimethylacetamide, and more preferably dimethyl sulfoxide, dimethylformamide, toluene or tetrahydrofuran.

2.1.1. Hexahydroxy small molecule initiator (IN- (OH)₆)

The structural general formula of the hexahydroxy micromolecule initiator is

Wherein A is₀、A₁、L₂The definitions of (A) are the same as those in the above general formula (1), and are not described herein again.

The hexahydroxy small molecule can be used as A through 1 residue₀The trifunctional small molecule and 3 residues can be A₁The trifunctional micromolecules are obtained through coupling reaction. Wherein, the residue may be A₀The trifunctional small molecule of (a) contains three identical functional groups; the residue may be a₁The tri-functional small molecule contains two or three identical functional groups, and the two ends of the tri-functional small molecule containing two identical functional groups are connected to A₁The atomic spacings of the branched nuclei being the same and consisting of A₁At least two ends of the branched core are identical, when the residue can be A₁When the tri-functional small molecule contains only two identical functional groups, the different functional group ends and residues can be A₀Are linked. The six terminal hydroxyl groups of the small molecule with six hydroxyl groups are almost same in activity, and the six hydroxyl groups are respectively connected with A₁The six-arm polyethylene glycol derivative further prepared by taking the small hexahydroxy molecule as an initiator molecule has the advantages of higher purity, more accurate control of molecular weight and distribution thereof in the product polymerization process, single product structure, no occurrence of structures of other multi-arm products, better performance, reduction of purification difficulty, reduction of the consumption of organic reagents in purification, reduction of cost and greenness and environmental protection.

The tri-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.

When preparing the hexahydroxy micromolecule initiator, the residue can be used as A₀、A₁The trifunctional small molecule of (A) is

Etc.; wherein j₂Selected from any one of 0, 1, 2, 3, 4, 5, 6; r₁Is a hydrogen atom, or is selected from any one of the following groups: 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_1-20Alkyl, substituted aryl, substituted C_1-20Open-chain heterohydrocarbyl, substituted heteroaromatic hydrocarbyl.

The structure of a specific hexahydroxy small molecule initiator is exemplified as follows:

wherein the integer j₂Is any one of 0, 1, 2, 3, 4, 5 and 6.

2.1.2. Initiator system

The structure of the hexahydroxy micromolecule can be characterized and confirmed by the 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 small hexahydroxy molecule 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 catalyzed acylation method, acetic anhydride-perchloric acid-dichloroethane catalyzed acylation method, acetic anhydride-N-methylimidazole-DMF catalyzed acylation method and the like), normal-temperature catalyzed 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 small hexahydroxy molecules are 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 diphenylmethyl potassium, more preferably metallic sodium, potassium or diphenylmethyl potassium, and most preferably diphenylmethyl potassium. 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, the quality is unstable, and the modification of the 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 low solubility in organic solvents (such as sodium methoxide, potassium methoxide, sodium hydride, potassium hydride, etc.) require a long 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 the deprotonation time is short and the deprotonation is incomplete, taking alkali as a nucleophilic reagent to participate in anionic polymerization to obtain low-molecular-weight impurities with target molecular weight of the target polymer chain being 0.5 times; 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 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, dimethylformamide or dimethylacetamide. In general, preference is given to working in aprotic solvents, preferably in dimethyl sulfoxide, dimethylformamide, toluene or tetrahydrofuran.

2.1.4. Terminating the polymerization reaction

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 is caused, active oxygen anions cause unstable product structure, and impurities with molecular weight larger than the target molecular weight are formed after being placed in 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 incomplete protonation of the product due to uneven stirring of the reaction system caused by an increase in viscosity or solidification. 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 10 minutes to 60 minutes, and the time control 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 preparation method of hexa-arm polyethylene glycol derivative relates to a hexa-functional micromolecule containing six functional groups

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.

2.2.1. Hexafunctionalized small molecules

The degradability of the hexafunctional small molecule is not particularly limited, and may contain only the stable linker STAG, or 1 or more than 1 degradable linker DEGG. Among them, the kind of the degradable linking group may be 1 or more.

The hexafunctional small molecule contains 6 identical reactive groups selected from suitable reactive groups in class a-class H of the present invention. The hexafunctional small molecule can be A through 1 residue₀The trifunctional small molecule and 3 residues can be A₁The trifunctional micromolecules are obtained through coupling reaction; wherein, the residue may be A₀The trifunctional small molecule of (a) contains three identical functional groups; the residue may be a₁The tri-functional small molecule contains two or three identical functional groups, and the two ends of the tri-functional small molecule containing two identical functional groups are connected to A₁The atomic spacings of the branched nuclei being the same and consisting of A₁At least two ends of the branched core are completely the same. When the residue may be A₁III of (2)When the functionalized small molecule only contains two identical functional groups, the different functional group end and residue can be A₀Are linked. The activity of the 6 terminal functional groups of the hexa-functionalized micromolecule is almost the same, the hexa-armed polyethylene glycol derivative can be endowed with accurate molecular weight and narrower molecular weight distribution in the subsequent coupling reaction with 6 linear polyethylene glycols, and the purification and separation difficulty can be reduced, the organic reagent dosage can be reduced, the cost can be reduced, and the hexa-functionalized micromolecule is more green and environment-friendly in the subsequent purification process.

In the preparation of hexafunctional small molecules, the residue may be A₀The tri-functional small molecule can be A except the residue in the preparation of the hexahydroxy small molecule initiator₀The tri-functional small molecule can also be

Etc.; the residue may be A₁The tri-functional small molecule can be A except the residue in the preparation of the hexahydroxy small molecule initiator₁The tri-functional small molecule can also be

And the like.

The hexafunctional small molecule is any one of the hexahydroxyl small molecule initiators IN- (OH) used IN the polymerization reaction described IN the above 2.1.1₆In addition, can also be selected from the following structuresOne of them is:

and the like.

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 hexafunctional small molecule and is coupled to form a divalent linking group L₂。

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. For general formula (1) there may be a PEG terminal hydroxyl group, a linear functionalized functional group (containing only one functional group) or a branched functionalized functional group (which may 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 of the functional groups mentioned above A-J, precursors including any of the reactive groups, variations thereof as precursors, substituted forms, protected forms, deprotected forms, 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, or 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. As another example, it is permissible for the terminal to contain two or more kinds of protected reactive groups, but in the subsequent application, when used for modification of a bio-related 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 target structure having the target functional group can be obtained by end-functionalizing the hexa-armed polyethylene glycol derivative obtained by the above-mentioned coupling reaction. 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 of class I-class J or derivatives thereof (such as targeting group, photosensitive group and the like), active ester group and maleimide group, active ester group and aldehyde group, active ester group and azide group, active ester group and alkynyl or protected alkynyl, active ester group and acrylate group, active ester group and methacrylate group, active ester group and acrylate group, maleimide group and azide group, maleimide group and alkynyl or protected alkynyl group, Maleimide and acrylate groups, maleimide and methacrylate groups, maleimide and acrylate groups, maleimide and carboxyl groups, maleimide and amino groups or protected amino or amine salts, maleimide and isocyanate groups, maleimide and protected thiol groups, aldehyde and azide groups, aldehyde and acrylate groups, aldehyde and methacrylate groups, aldehyde and acrylate groups, aldehyde and epoxy groups, aldehyde and carboxyl groups, aldehyde and alkyne groups or protected alkyne groups, azide and thiol groups or protected thiol groups, azide and amino groups or protected amino or amine salts, azide and acrylate groups, azide and methacrylate groups, azide and acrylate groups, azide and carboxyl groups, acrylate groups and amino groups or protected amino or amine salts, acrylate groups and isocyanate groups, Acrylate group, epoxy group and acrylate group Acrylate and methacrylate group, acrylate and carboxyl group, methacrylate and amino group or protected amino or amine salt, methacrylate and isocyanate group, methacrylate and epoxy group, alkynyl or protected alkynyl and amino group or protected amino 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, acrylate and isocyanate group, acrylate and acrylate group, acrylate and epoxy group, acrylate and carboxyl group, carboxyl and mercapto group or protected mercapto group, Carboxyl and amino or protected amino or amine salt, carboxyl and isocyanate group, carboxyl and epoxy group, amino or protected amino or amine salt and sulfhydryl or protected sulfhydryl, 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 ₂HCl。

The bilmpeg may be polydisperse or monodisperse. When polydisperse, corresponds to n₁≈n₂≈n₃≈n₄≈n₅≈n₆When monodisperse, corresponds to n₁＝n₂＝n₃＝n₄＝n₅＝n₆。

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 obtained product, and when used for modifying molecules such as drugs, the higher the quality of the modified product, the more practical requirements can be met.

When the bilmpeg is monodisperse, PDI is 1, six PEG chains each have a fixed molecular weight and are equal to each other, and a six-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 the monodisperse polyethylene glycol chain can adopt the well-known technology in the technical field, including but not limited to the document J.Org.chem.2006,71, 9884. sup.9886 and the cited document thereof, the document Angew.chem.2009,121,1274-1278 and the cited document thereof, the document Expert Rev.mol.Diagn.3, 13(4), 315. sup.319 and the cited document thereof, the document Angew.chem.int.Ed.2014,53, 6411. sup.6413 and the cited document thereof, the document Bioorganic & Medicinal Chemistry Letters 2015,25:38-42 and the cited document thereof, the document Angew.m.int.Ed.2015, 54: 3763. sup.3767 and the applied document thereof, and the like.

2.3. A preparation method of six-arm polyethylene glycol derivatives relates to 1 trifunctional micromolecule containing three same functional groups and 3 branched polyethylene glycol molecules V-PEG₂The coupling reaction process of (1), reacting to obtain a hexa-arm polyethylene glycol derivative; wherein, the branched V-PEG₂Containing two identical branched PEG chain segments and one end of a main chain functional group derived from a branched core, the functional groups of the two PEG chain ends and the main chain end may be the same or different, and V-PEG₂The main chain functional group end of the molecule is connected with the trifunctional micromolecule; wherein, V-PEG₂Is monodisperse or polydisperse;

2.3.1. Trifunctionalized small molecules

The residue in the preparation of the hexafunctional micromolecule can be A₀The trifunctional small molecules of (a) are not described in detail herein.

2.3.2. Branched polyethylene glycol molecule V-PEG₂

Branched V-PEG₂Contains two identical branched PEG chain segments and one main chain non-PEG chain functional group end led out from the branched core, the functional groups of the two PEG chain ends and the main chain end can be identical or different, and the V-PEG₂The main chain functional group end of the molecule is connected with the trifunctional micromolecule, and then the V-PEG is added₂The branched core of the molecule is A₁The branched core of (3).

Branched polyethylene glycol molecule V-PEG₂The functional groups of the two PEG chain ends are the same or different from the target structure, and the functional groups may be the same or different from the target functional groups. For general formula (1) there may be a PEG terminal hydroxyl group, a linear functionalized functional group (containing only one functional group) or a branched functionalized functional group (which may 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 of the functional groups mentioned above A-J, precursors including any of the reactive groups, variations thereof as precursors, substituted forms, protected forms, deprotected forms, 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, or 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. As another example, it is permissible for the terminal to contain two or more protected reactive groups, but in subsequent applications, when used to modify a biologically relevant substance, only one of them is selectively deprotected A single kind of reactive group is obtained. When the structure and the functional group at the end are different from the target, the target structure having the target functional group can be obtained by end-functionalizing the hexa-armed polyethylene glycol derivative obtained by the above-mentioned coupling reaction. 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.

Branched V-PEG₂The functional groups at the ends of the two PEG chains and the functional groups at the ends of the main chain of (A) may be the same or different, and branched V-PEG having two different functional groups is preferred₂Derivatives, the number of two different functional groups being one of 1 and the other two, wherein the pair of heterofunctional groups which may be present simultaneously is as described in section 2.2.2.

Branched V-PEG₂It may be polydisperse or monodisperse. When polydisperse, corresponds to n₁≈n₂≈n₃≈n₄≈n₅≈n₆When monodisperse, corresponds to n₁＝n₂＝n₃＝n₄＝n₅＝n₆。

When branched V-PEG₂When it is a polydispersity, the 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 preferred. The lower the PDI, the more uniform the molecular weight, and the narrower the molecular weight distribution of the obtained product, and when used for modifying molecules such as drugs, the higher the quality of the modified product, the more practical requirements can be met.

When branched V-PEG₂In the case of monodispersity, PDI ═ 1, six PEG chains each have a fixed molecular weight and are equal to each other, and a six-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.

2.4. 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 groups include, but are not limited to, the functional groups listed in class A through 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 ₀₁The 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 branching group with a valence of k +1, and a functional group R at the tail end of a polyethylene glycol chain₀₁The 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.

2.4.1. 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 hydroxyl group at the end of the polyethylene glycol chain, or conversion to the target functional group based on the 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 reaction types involved (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.

2.4.1.1. Class A: r₀₁Functionalization 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 the terminal hydroxyl intermediate with the corresponding carbonates (N, N '-disuccinimidyl carbonate, di (p-nitrophenyl) carbonate, di (o-nitrophenyl) carbonate, bis-benzotriazol carbonate, etc.), haloformates (p-nitrophenyl chloroformate, o-nitrophenyl chloroformate, trichlorophenyl chloroformate, etc.), N' -carbonyldiimidazole in the presence of a base. The corresponding ring-substituted derivatives of hydrogen atoms can also be obtained in an analogous 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 is preferably 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. The solvent may be a non-solvent or an aprotic solvent. The base includes generally organic bases, preferably triethylamine, pyridine.

2.4.1.2. Class B: r₀₁Functionalization selected from class B

The sulfonic acid or sulfinate derivatives (B1, B2) can be reacted with a leaving group Y through a terminal hydroxyl group₁The sulfonyl chloride and sulfinyl chloride are obtained by esterification in the presence of alkali. Y is ₁The definitions of (a) and (b) are consistent with the above. 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.

The sulfone or sulfoxide derivative (B3, B4) can be modified by containing a leaving group Y₁The sulfoxide intermediate or thioether intermediate is prepared by oxidation reaction. Y is₁The 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 }.

2.4.1.3. Class C: r₀₁Functionalization 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 preferably water, toluene, benzene, xylene, acetonitrile, ethyl acetate, diethyl ether, methyl tert-butyl ether, tetrahydrofuran, chloroform, dichloromethane, dimethyl sulfoxide, dimethylformamide or dimethylacetamide, and 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 the 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 an organic base or an inorganic base, preferably an organic base, 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 the imine (Schiff base) is reduced to the corresponding alkylated amine compound (C5) in the presence of a reducing agent after the imine has been prepared by reacting the amine with the corresponding aldehyde or ketone. 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), tetramethylpiperidinyloxy compound (C8), and dioxopiperidinyloxy compound (C9) can be obtained by reacting the sulfonate compound (B1) with the corresponding halide salt, 2,6, 6-tetramethylpiperidine-nitrogen-hydroxy, 3, 5-dioxo-1-cyclohexylamine. The bromine salt is not limited as long as free bromide ions are 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 compound (C17) can be obtained by condensation 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- (benzylthio-thiocarbonylthio) 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 with corresponding alcohol (H1) or amine (C4).

2.4.1.4. Class D: r₀₁Functionalization 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. For example, ethyl chloroacetate and ethyl bromoacetate.

Thioesters (D26) are also obtained by reaction of the corresponding ester (D11) with a thiol.

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_1-10Acid chloride, C_1-10Acyl bromide, C_1-10Anhydride, 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 used₂Preferably 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, and sodium, potassium, sodium hydride, potassium hydride, sodium methoxide, potassium tert-butoxide, or diphenylmethyl potassium is preferred, and sodium hydride or diphenylmethyl potassium is more preferred. 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, and removing acetal protection₂)_0～3A 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 _1-10Diisocyanate, C_1-10A 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 di-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 basic conditions. 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.

2.4.1.5. Class E: r₀₁Functionalization 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-maleimidopropionic 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) butyrate 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. The amine 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 then reacting with the 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 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 preferred, and DCC is most preferred. 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 compounds 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-methylamine, 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).

2.4.1.6. Class F: r₀₁Functionalization selected from class F

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.; epichlorohydrin is preferred. 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-tert-butyldimethylsilylbromidepyne.

2.4.1.7. Class G: r₀₁Functionalization 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, urethane bonds, carbonate bonds, hydrazide bonds and the like. The following are exemplified as raw materials:

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 amine derivative (C4) is obtained by firstly pressurizing with ammonia and then pressurizing with hydrogen under the catalysis of nickel or palladium carbon, and then carrying out dehydrogenation reaction under the condition of high temperature.

Compounds (G31) and (G32) can be prepared using the methods of the literature PCT/US2013/046,989.

2.4.1.8. Class H: r₀₁Functionalization 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) CH₂CH₂An 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 halosilane, 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 reacted with a halosilane, acid chloride, acid anhydride, sulfonyl chloride, halohydrocarbon under neutral conditions or in the presence of a base to afford 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 ₄In 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)₀₁The 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. Among them, 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 base, vinyl ether base, dihydropyran base, benzyl and Boc.

The end-protected bishydroxy group (H3) can be obtained by a method including, but not limited to, references { Macromol. biosci.2011,11,1570-1578}, 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.

2.4.1.9. Class I: r₀₁Functionalization 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, and DCC are preferred. 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 terminal hydroxyl group of polyethylene glycol with carboxylic acid (D1), acid halide (D4), sulfonyl chloride (D5), isocyanate (D9), isothiocyanate (D10), and the like. The pegylated cholesterol may also be obtained by coupling of a derivative of cholesterol with a suitable group. Taking cholesteryl hydrogen succinate as an example, the cholesteryl hydrogen succinate 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 suitable polyethylene glycol or derivatives thereof 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.

2.4.1.10. Class J: r₀₁Functionalization 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), and succinimide active esters (A1, A6), carboxyl (D1), primary amino (C4), secondary amino (C15), hydrazine or substituted hydrazine (C12), such as N-aminocarbazole, cyano (G23), unsaturated bonds of maleimide (E1), maleimide (C21), aldehyde groups (D6), acrylate groups (E2), methacrylate groups (E3), oxime groups (G24), and derivatives (J3957), 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.

2.4.1.11. Conversion to target functional groups based on reactive groups

The method can be realized by any one of the following modes: (1) and (3) direct modification, namely, based on the direct modification of the reactive group, obtaining the target functional group. 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 methods 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, a condensation reaction to form an ester group, a thioester group, an amide group, an imine linkage, a hydrazone linkage, a carbamate group, 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, cyanooxide and the like and the target functional group as raw materials. 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 are 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.

2.4.2. 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. At this time, 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.

2.4.2.1. 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 steps 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 L₀. 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 ether bond is formed, at which time L ₀Comprises CH₂CH₂O; for another 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. forming a new bond NHCOO, COO, etc. is contained in L₀Performing the following steps; 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 includesFunctional modifications at the hydroxyl group also include conversion to a new functional group based on a non-hydroxyl functional group.

The method for introducing the above-mentioned branched group is not particularly limited, and the existing techniques in the chemical field may be employed as long as the covalent bonding can be formed, including but not limited to the various coupling reactions described above. Examples include Macromolecules 2013,46,3280-3287, Macromolecules 2012,45,3039-3046, Macromolecules Chem 2012,3,1714-1721, US5,811,510, US7,790,150, US7,838,619, etc., and examples include Journal of Polymer Science, Part A, Polymer Chemistry,2013,51,995-1019, macromolecule Biosci.11, 11, 3-propan-2, hyperbranched structures 1562, 2011-rapid-mut 2010,31,1811-1815, Langir 2010,26(11), 2011-8881, hyperbranched structures 2011-d-mut.2010, 31,1811-1815, Langir 2010, 2633, 75, 250-8881, etc., hyperbranched structures contained in Macromolecules, 2011-883, 2011-250, 2703-250, 2705, Polyporacle-250-wo 75, 2703-250-20-one-wo 29, 2011, 2673, 2633, 2673, 2703-73, 2673, a. The branched structures described in the above documents and the methods for their preparation 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.

2.4.2.2. Terminal branched functionalized feedstock

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

) 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, two protected primary amines or two protected secondary amines, thiols, 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 (dihydroxy)Monocarboxylic acids) include, but are not limited to, 2, 3-dihydroxypropionic acid, 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.

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, hydroxy dicarboxylic acids (including but not limited to tartronic acid, L-malic acid, D-malic acid, citramalic acid, 3-hydroxyglutaric acid), amino dicarboxylic acids (including but not limited to 2-aminomalonic acid, diethyl 2-aminomalonate, 3-aminoglutaric acid), mercapto dicarboxylic 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, fumaric acid, malic acid, fumaric, 4-amino-2- (2-aminoethylamino) butyric acid with two amino groups protected, glycerol dimethacrylate, 2-bis (allyloxy) butyric acid Methyl) -1-butanol,

And the like, and a form in which any of the above-mentioned functional groups in an amount of 2 is protected.

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 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 surfactant 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 heteropentafunctional 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 forms of any of the foregoing in which the number of functional groups is 5 is protected .

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

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-hydroxyiminolevulinic acid, N-hydroxybutanedioic acid, N-hydroxyiminolevulinic acid, N-,

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 or 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,

Ethyl-3-oxetanylcarbinol),

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 two hydroxyl groups are protected (e.g., monophenylol pentaerythritol, forming polypentaerythritol), and mixtures thereof,

(F is as defined above, preferably in protected form, one of the preferred forms being protected hydroxy OPG₄Such as 1-ethoxyethyl-2, 3-epoxypropyl ether, benzyl glycidyl ether, tert-butyl glycidyl ether, allyl glycidyl ether, propargyl glycidyl ether, glycidyl methacrylate, glycidyl ether,

(e.g., azidopropyl methacrylate), carbon dioxide and

combinations of (e.g. { Macromolecules 2013,46,3280-3287}, and further e.g. carbon dioxide with a compound selected from the group consisting of

Propargyl glycidyl ether, and the like), a combination of a diisocyanate and a diol containing 1 reactive group or a protected form thereof,

Combinations with diamines (forming combs with multiple thiol groups suspended, { Macromol. RapidCommun.2014,35,1986-1993}), D-glucopyranose units (forming acetalized glycan structures such as poly (1 → 6) hexose, 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, and the like. Other trihydric alcohols, trihydric alcohols containing one protected hydroxyl group, tetrahydric alcohols containing 2 protected hydroxyl groups, and polyhydric alcohols containing 2 naked hydroxyl groups and other protected hydroxyl groups can be used as raw materials for preparing the comb-shaped structure. 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.

2.4.2.3. 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

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),

And the phosphorus atom branching center can be obtained by taking phosphoric acid, phosphate ester, thiophosphoric acid and thiophosphate as raw materials.

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 yield trivalent nitrogen branching centers. As another example, reaction between an alkynyl group and 2 molecules of a thiol group 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.

2.5. 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 form a covalent linking group upon reaction. The reaction conditions, depending on the type of covalent linking group formed by the reaction, can be as described in the prior 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 of the covalent linking group may be divalent or trivalent, with divalent predominating.

The coupling reaction can produce stable groups as well as degradable groups.

In general terms, for example: the amino group reacts with active ester, formic acid active ester, sulfonic ester, aldehyde, alpha, beta-unsaturated bond, carboxylic acid group, epoxide, isocyanate and isothiocyanate to obtain amide group, urethane group, amino group, imino group (which can be further reduced into secondary amino group), and ammonia Divalent linking groups such as a group, an amide group, an amino alcohol, a urea bond, and a thiourea bond; reacting a sulfhydryl group with a divalent linking group containing an active ester, a formic acid active ester, a sulfonic ester, a sulfhydryl group, maleimide, aldehyde, an alpha, beta-unsaturated bond, a carboxylic acid group, iodoacetamide and an anhydride to obtain a thioester group, a thiocarbonate, a thioether, a disulfide, a thioether, a hemithioacetal, a thioether, a thioester, thioether, imide and the like; 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 linking 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

Also as the following diamines and aldehydesReaction of radicals

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 may be carried out in a manner 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; (3) the method 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 ₂NHCH₂-) can be obtained by condensation and reduction reaction between aldehyde group and amino group, or by alkylation reaction between primary amine and sulfonate 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-dihydroisoxazole is formed, the compoundThe product is obtained by 1, 3-dipolar cycloaddition reaction between peroxycyanide and alkynyl.

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 sulfonates or halides, which in turn correspond to the formation of ether linkages, thioether linkages, secondary or tertiary amino groups.

2.6. 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 six-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 each of the cited documents. The terminal functionalization rate (substitution rate) of the hexa-arm polyethylene glycol derivative, namely the mole percentage of the terminal hydroxyl group of the hexa-arm polyethylene glycol which is functionalized and modified, is determined by the terminal hydroxyl group characteristic peak-CH in the hexa-arm polyethylene glycol raw material nuclear magnetism characteristic peak ₂CH₂OH and EO Block characteristic Peak-CH₂CH₂Characteristic peak of functional group and characteristic peak of EO block-CH in nuclear magnetism characteristic peak of O-, and hexa-arm polyethylene glycol derivative products₂CH₂The integral ratio of O-is obtained by conversion, and the conversion method is well known to those skilled in the art and is not described in detail herein.

2.7. 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 biological related substance modified by the six-arm polyethylene glycol derivative, which has the following structural general formula:

EF may be expressed as ED (structure is

) Or EF₁(structure is

) And D is not equal to E₀₁(ii) a Wherein q and q are₁Each independently is 0 or 1; z₁、Z₂Each independently is a divalent linking group; wherein D is a residue formed by the reaction of the modified bio-related substance and the hexa-arm polyethylene glycol derivative; e₀₁Is selected from R₀₁Protected R₀₁Deprotected R ₀₁Or blocked R₀₁；R₀₁Is a reactive group capable of reacting with a biologically relevant substance; l is a linking group formed after a reactive group in the six-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_DK is not more than k, k of each branched chain in the same molecule_DEach independently the same or different, and the sum of the numbers of D in any one of the hexa-armed polyethylene glycol derivative molecules (N)_D) At least 1, preferably at least 6; when G is 1, G- (EF)_kCan be expressed as

The biologically-relevant substance preferably has a plurality of reaction sites, and the same biologically-relevant substance and the same R₀₁The residues D obtained by the reaction may be the same or different;

In the general formula (2), when g is 0, the structure of the bio-related substance modified with the hexa-arm polyethylene glycol derivative is represented by the formula (3).

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 bio-related substance modified by the hexa-arm polyethylene glycol derivative is shown as a formula (4).

K at the end of six PEG chains in the same molecule_DPreferably all satisfy 1. ltoreq. k_DK, i.e.at least one D is attached to each branch chain. Ideally, k is at the end of six PEG chains in the same molecule_DAll satisfy k_DK, 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 R₀₁The 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 site) in a single molecule is not particularly limited and may be largeAt about 75%, equal to about 75%, or less than about 75%. The macroscopic material constituting the product of the six-arm polyethylene glycol derivative modification may have the same or different D content in each molecule, such as 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 ₀₁After 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% and also 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_DThe 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 one average six-arm polyethylene glycol derivative molecule can be an integer or a non-integer; wherein the integer k in a single molecule_DEach independently is 0, 1 or 2 to 250. The invention also covers the substances of one molecule of the biologically relevant substance combined with 2 or more molecules of the hexa-armed polyethylene glycol derivative, but preferably 1 molecule of the biologically relevant substance reacts with only 1 functional group, namely only one molecule of the hexa-armed polyethylene glycol derivative is connected, and the corresponding quality controllability is strong. I.e. k _DAlso indicated is the number of bio-related substance molecules bound in F (expressed as a mean value for macroscopic substances, i.e. the number of bio-related substances to which one hexa-armed polyethylene glycol derivative molecule is attached on average). The functional group modified by the hexa-arm polyethylene glycol derivative can be totally or partially involved in the modification of the biological related substances. Preferably all involved in the modification of the biologically relevant substance. In the bio-related substance modified by the six-arm polyethylene glycol derivative, the functional group which is not bonded with the bio-related substance can be in a structural form before reaction, a protected form, a deprotected form or a non-bio-related substance end capping.

L may be covalently or non-covalently attached. Preferably a covalent linker; it may also be a double or multiple hydrogen bond. Since reactions with different sites from the same biologically relevant substance are allowed, the six PEG chain ends of the same molecule are allowed to correspond to different L, preferably the L at the six PEG chain ends is the same. Any one of L is independently stable or degradable, and the linking 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)_qEach L is independently stably present or degradable, and (Z)₂)_qThe linking group of-L to the adjacent heteroatom group may be stable or degradable. Preferably, the L's at the six PEG chain ends have the same stability, i.e., are all stably present or all degradable, in which case (Z) at the six PEG chain ends₂)_q-L also has the same stability.

The difference of the stability property (also called degradability) of the bio-related substance modified by the hexa-arm polyethylene glycol derivative according to the degradation position includes but is not limited to the following cases:

(1) g-0, having stable hexavalent centers

stabilized-O- (Z)₂)_q-L-；

(2) g-0, having stable hexavalent centers

degradable-O- (Z)₂)_q-L-；

(3) g-0, having degradable hexavalent centers

degradable-O- (Z)₂)_q-L-；

(4) g 1, having stable hexavalent centers

stabilized-O-L₀-G- (G and Z are excluded)₂Connection of), stable- (Z)₂)_q-L- (containing G and Z)₂The connection of (c);

(5) g 1, having stable hexavalent centers

degradable-O-L₀-G- (G and Z are excluded)₂Connection of), stable- (Z)₂)_q-L- (containing G and Z)₂The connection of (c);

(6) g 1, having stable hexavalent centers

stabilized-O-L₀-G- (G and Z are excluded)₂Linked) degradable- (Z)₂)_q-L- (containing G and Z)₂The connection of (c);

(7) g 1, having degradable hexavalent centers

(8) g 1, having degradable hexavalent centers

(9) g 1, having degradable hexavalent centers

(10) g 1, having stable hexavalent centers

degradable-O-L₀-G- (G and Z are excluded)₂Linked) degradable- (Z)₂)_q-L- (containing G and Z)₂The connection of (c);

(11) g 1, having degradable hexavalent centers

(12) g-0, having stable hexavalent centers

stabilized-O- (Z)₂)_q-, degradable L;

(13) g 1, having stable hexavalent centers

stabilized-O-L₀-G-[(Z₂)_q-]_kA degradable L;

(14) g-0, having stable hexavalent centers

stabilized-O- (Z)₂)_qDegradable L-D;

(15) g 1, having stable hexavalent centers

stabilized-O-L₀-G-[(Z₂)_q-]_kCanDegraded 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), (13) correspond to stable six-arm polyethylene glycol derivative; (3) degradable six-arm polyethylene glycol derivatives corresponding to the items (5), (7) and (11); (2) provided that (Z) in (1) and (6) ₂)_q- (containing Z)₂Attachment to the PEG terminus O or G) and L) may be degradable, and thus may correspond to a stable hexa-armed polyethylene glycol derivative containing degradable L, or may correspond to a degradable hexa-armed polyethylene glycol derivative containing stable L. One preference of the above combination is to have stable hexavalent centers

Comprises (1), (2), (4), (5), (6), (10), (12), (13), (14) and (15). Wherein, for the two cases (1) and (4), the PEG part is not degradable, and the connection between the PEG part and the bio-related substance also exists stably, and the bio-related substance modified by the polyethylene glycol derivative can be a modified bio-related 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 passed₁)_q1-R₀₁Can 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 six PEG chains in the general formula (2) are from the same source, so the lengths of the six PEG chains are completely the same or close to each other, and the biologically-relevant substances modified by the six-arm polyethylene glycol derivatives 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 n₁＝n₂＝n₃＝n₄＝n₅＝n₆N corresponds to a monodisperse PEG chain, or n₁≈n₂≈n₃≈n₄≈n₅≈n₆N 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 and modification products generated by additionally introducing structures such as functional groups, reactive groups, amino acids or amino acid derivatives, polypeptides and the like belong to chemical modification substances of biological related substances. The bio-related substance may also allow for a target molecule, adjunct or delivery vehicle to bind to it, either before or after binding to the hexa-armed polyethylene glycol derivative, 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, 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 medical bio-related substances including but not limited to drugs, drug carriers and medical devices can be used for 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 disease and the like, anti-allergic 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, alzheimer's drugs or compounds, Imaging agents, antidotes, anticonvulsants, muscle relaxants, anti-inflammatory agents, appetite suppressants, migraine agents, muscle contractants, antimalarials, antiemetics/antiemetics, bronchodilators, antithrombotic agents, antihypertensive agents, antiarrhythmics, antioxidants, anti-asthma agents, diuretics, lipid regulating agents, antiandrogens, antiparasitics, anticoagulants, anti-neoplastic agents, hypoglycemic agents, nutritional agents, additives, growth supplements, anti-enteritis agents, vaccines, antibodies, diagnostic agents (including but not limited to contrast agents), contrast agents, hypnotics, sedatives, psychostimulants, tranquilizers, anti-parkinson agents, analgesics, anxiolytics, muscle infectives, auditory disease agents, 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, rhabdomyosarcoma, sarcoma, and colon cancer, Primary or secondary carcinoma, sarcoma or carcinosarcoma such as neuroblastoma, AIDS-related cancer (such as Kaposi's sarcoma).

"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, lipids, hormones, vitamins, vesicles, liposomes, phospholipids, glycolipids, dyes, fluorescent substances, targeting factors, cytokines, neurotransmitters, extracellular matrix substances, 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 six-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. The reactive sites of the amino acids described in the literature { Journal of Controlled 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 ₀₁Examples 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 or 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; reacting a biological related substance containing sulfydryl with polyethylene glycol containing active ester, formic acid active ester, sulfonate, 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, thiohemiacetal, 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; respectively reacting biologically-relevant substances containing carbonyl or aldehyde groups 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

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 can stably exist under light, heat, low temperature, enzyme, redox, acidic, basic, physiological conditions or in vitro simulated environments, or a linker that can degrade under light, heat, low temperature, enzyme, 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 a degradable linking group including, but not limited to, any of the degradable linking groups described above, 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, acetals, cyclic acetals, mercaptals, azaacetals, azaheterocyclic acetals, dithioacetals, hemiacetals, thiohemiacetals, azahemiacetals, ketals, thioketals, azaheterocyclic ketals, thioketal, imine bonds, hydrazone bonds, acylhydrazone bonds, oxime bonds, thiooxime bonds, semicarbazone bonds, thiosemicarbazone bonds, hydrazine groups, hydrazide groups, thiocarbhydrazide groups, azohydrazide groups, thiohydrazide groups, thioazocarbohydrazide groups, thiohydrazide groups, Hydrazinoformate groups, hydrazinothiocarbamate groups, carbazide, thiocarbohydrazide, azo groups, isoureido groups, isothioureido groups, allophanate groups, thioallophanate groups, guanidino groups, amidino groups, aminoguanidino groups, amidino groups, imino groups, thioester groups, sulfonate groups, sulfinate groups, sulfonamide groups, sulfonyl hydrazide groups, sulfonyl urea groups, maleimide groups, orthoester groups, phosphate groups, phosphite groups, hypophosphite groups, phosphonate groups, phosphosilane groups, silane groups, carbonamide, thioamide, phosphoramide, phosphoramidite, pyrophosphoroamide, cyclophosphamide, ifosfamide, thiophosphoramide, aconityl groups, peptide bonds, thioamide bonds and 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 a 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 hexa-armed polyethylene glycol derivatives and biologically related substances

Reactions between hexa-armed 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 hexa-armed polyethylene glycol derivative and the biologically-relevant substance is not particularly limited, and may be a site-directed modification or an undefined site modification (also referred to as a random modification). By way of example, site-directed modifications such as commercial products

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, vinylsulfone, 2-iodoacetamide, o-pyridyldisulfide, 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

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 hexa-arm polyethylene glycol derivative modifies the bio-related substance, 1 or more than 1 hexa-arm polyethylene glycol derivative molecules can be connected to one bio-related substance. For reference, e.g. commercial products

One molecule of polyethylene glycol reacts with only one reaction site in one drug molecule; to commercialize the product

In this case, one drug molecule may be linked to a plurality of polyethylene glycol molecules. It is preferred in the present invention that a biologically relevant substance is bound to only one hexa-armed polyethylene glycol derivative molecule.

When the hexa-arm polyethylene glycol derivative modifies a bio-related substance having two or more reaction sites, the hexa-arm polyethylene glycol derivative 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 hexa-arm polyethylene glycol derivative without special description; preferably, 1 molecule of the biologically relevant substance reacts with only 1 functional group.

3.4. Micromolecular drug modified by six-arm polyethylene glycol derivative

The invention also discloses a micromolecular drug modified by the six-arm polyethylene glycol, 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, organic metal compound, oligopeptide or polypeptide 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 the biologically relevant 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 mimicry or active fragments (including variants) of any biologically relevant 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 analog, a mimetic, a polymorph, a pharmaceutically acceptable salt, a fusion protein, a chemically modified substance, a gene recombinant substance, and the like thereof, or a corresponding agonist, activator, inhibitor, antagonist, modulator, receptor, ligand or ligand, 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 by way of example but not limited to anticancer drugs, antineoplastic drugs, anti-hepatitis drugs, diabetes treatment drugs, anti-infective drugs, antibiotics, antiviral agents, antifungal agents, vaccines, anti-respiratory drugs, anti-spasmodics, muscle relaxants, anti-inflammatory drugs, appetite suppressants, migraine treating agents, muscle contractants, antirheumatics, antimalarials, antiemetics, bronchodilators, antithrombotic agents, antihypertensive agents, cardiovascular agents, antiarrhythmic agents, antioxidants, anti-asthmatic agents, diuretics, lipid regulating agents, antiandrogenic agents, antiparasitic agents, anticoagulants, anti-neoplastic agents, hypoglycemic agents, nutritional agents and additives, growth supplements, anti-enteritis agents, antibodies, diagnostic agents, anti-cancer drugs, anti-cancer, 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 SN38, irinotecan and resveratrolCantharidin and its derivatives, Buxus sinica Linn, radix Tripterygii Wilfordii extract, flavone or flavonoid drug, Saviae Miltiorrhizae radix extract, and herba Silybi Mariani extract or their derivatives or their pharmaceutically acceptable salts; 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, polypeptide derivatives, in addition to the molecularly modified derivatives. When the hexa-armed polyethylene glycol derivative is combined with the small-molecule drug through alcoholic hydroxyl group or phenolic hydroxyl group, the amino acid derivative of the small-molecule drug or the oligo-polyethylene glycol fragment with 2-10 EO units is preferred, the amino acid derivative of the small-molecule drug is more preferred, the glycine-modified product of the small-molecule drug is most preferred, namely, the L preferably contains an amino acid derivative skeleton, the glycine skeleton or the alanine skeleton is more preferred, and the 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 hexa-armed polyethylene glycol derivatives and the preparation 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 these examples, the monodisperse starting materials, key intermediates and products were prepared by preparing hexa-armed polyethylene glycol and hexa-armed polyethylene glycol derivatives, and the molecular weights were confirmed 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 six-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 atom branched center hexahydroxy micromolecule S1-3

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(29.28g, 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 hexahydroxy micromolecule initiator S1-3 is obtained through concentration, washing and column chromatography purification. The hydrogen spectrum data of the hexahydroxy small molecule S1-3 are as follows: ¹H NMR(DMSO-d₆)δ(ppm):2.52-2.56(N(CH₂CH₂O-)₃,6H),3.15-3.25(-OCH(CH₂OH)₂,3H),3.32-3.48(-CH(CH₂OH)₂,12H),3.48-3.52(N(CH₂CH₂O-)₃,6H)。

Example 2: preparation of six-hydroxyl micromolecule S2-3 with carbon atom branching center

The preparation process is as follows:

adding 400mL of tetrahydrofuran, a compound S2-1(1.20g, 10.0mmol) and excessive diphenyl methyl potassium (75.0mmol) into an anhydrous and oxygen-free closed reaction kettle, then adding a compound S2-2(33.63g, 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, after the reaction is carried out for 4 hours, the hexahydroxy micromolecule initiator S2 is obtained by concentration, washing and column chromatography purification-3. The hydrogen spectrum data of the hexahydroxy small molecule S2-3 are as follows:¹H NMR(CDCl₃)δ(ppm):0.98(CH₃C(CH₂O-)₃,3H),2.60-2.70(>NCH₂CH₂O-,6H),2.75-2.85(-N(CH₂CH₂OH)₂,12H),3.37(CH₃C(CH₂O-)₃,6H),3.48-3.68(>NCH₂CH₂O-,6H),3.76-3.80(-N(CH₂CH₂OH)₂,12H)。

example 3: preparation of hexahydric micromolecule S3-3 with silicon atom as branching center

The preparation process is as follows:

into an anhydrous and oxygen-free closed reaction kettle, 400mL of tetrahydrofuran, the compound S3-1(0.94g,10.0mmol) and excess diphenylmethyl potassium (75.0mmol) are added successively, followed by

(S3-2, 33.63g, 75.0mmol), 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 hexahydroxy micromolecule initiator S3-3 is obtained through concentration, washing and column chromatography purification. The hydrogen spectrum data of the hexahydroxy small molecule S3-3 are as follows: ¹H NMR(CDCl₃)δ(ppm):0.01(SiCH₃,3H),1.28(-CCH₃,3H),2.55-2.65(>C(CH₂CH₂OH)₂,12H),3.76-3.85((>C(CH₂CH₂OH)₂,12H)。

Example 4: preparation of hexahydroxy micromolecule S4-2 at cyclohexane branching center

The preparation process is as follows:

adding 400mL of tetrahydrofuran, a compound S4-1(1.32g, 10.0mmol) and excessive diphenyl methyl potassium (75.0mmol) into an anhydrous and oxygen-free closed reaction kettle, then adding a compound S1-2(29.28g, 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 hexahydroxy micromolecule initiator S4-2 is obtained through concentration, washing and column chromatography purification. The hydrogen spectrum data of the hexahydroxy small molecule S4-2 are as follows:¹H NMR(CDCl₃)δ(ppm):1.23-2.11(>CHCH₂CH<,6H),3.40-3.55(>CHOCH(CH₂OH)₂,3H；-CH(CH₂OH)₂,3H),3.62-3.78(-CH(CH₂OH)₂,12H)。

example 5: preparation of nitrogen atom branched center six-arm polyethylene glycol butyne derivative E1-1

Corresponding to the general formula (1), wherein A₀Is a nitrogen atom

A₁Is composed of

L_A0A1Containing an ether bond of-CH₂CH₂O-，L₂is-CH₂-，F_GIs composed of

The designed total molecular weight is about 9852Da, wherein each PEG chain has a molecular weight of about 1500Da, corresponding to n₁≈n₂≈n₃≈n₄≈n₅≈n₆≈34。

The preparation process is as follows:

step a: tetrahydrofuran (500mL), a hexahydroxy small-molecule initiator S1-3(0.93g,2.5mmol) and diphenylmethyl potassium (6.0mmol) were added sequentially to a water-free and oxygen-free closed reaction kettle.

Step b: adding calculated amount of ethylene oxide, gradually raising the temperature to 60 ℃, and reacting for 48 hours.

Step c: an excess of methanol, a proton source, was added to give a six-arm polyethylene glycol H1-1 with a nitrogen atom as a branching center. The hydrogen spectrum data of H1-1 is as follows:¹H NMR(CDCl₃)δ(ppm):3.33-3.52(-CH(CH₂O-)₂,12H),3.55-3.75(-OCH₂CH₂OH,24H；-OCH₂CH₂O-)。

step d: in a dry clean 1L round bottom flask, hexa-armed polyethylene glycol H1-1(18.74g,2.0mmol), excess 4-pentynoic acid (3.53g,36.0mmol) and solvent dichloromethane (200mL) were added, DMAP (0.05g,0.4mmol) was added under ice bath, DCC (7.40g,36.0mmol) dissolved in 100mL dichloromethane was added dropwise to the reaction solution, and after the addition was completed, the reaction was carried out at room temperature for 16H. After completion of the reaction, insoluble matter was removed by filtration, concentrated and purified by column chromatography to give a six-arm polyethylene glycol butyne derivative E1-1(15.37g, yield 78%) having a nitrogen atom as a branching center. The hydrogen spectrum data of E1-1 is as follows:¹H NMR(CDCl₃)δ(ppm):2.00(-CH₂CH₂C≡CH,6H),2.42-2.62(-OC(O)CH₂CH₂C≡CH,24H),3.33-3.52(-CH(CH₂O-)₂,12H),3.55-3.75(-OCH₂CH₂OC(O)-,12H；-OCH₂CH₂O-),3.90-4.20(-OCH₂CH₂OC(O)-,12H)。

to six-arm polyethylene glycolButyne derivative E1-1 was subjected to GPC measurement to determine M_n≈9.9kDa,PDI＝1.02。

Example 6: preparation of six-arm polyethylene glycol azide derivative E2-1 with carbon atom as branching center

Corresponding to the general formula (1), wherein A₀Is composed of

A₁Is composed of

L_A0A1Containing an ether bond of-CH₂OCH₂CH₂-，L₂is-CH₂CH₂-，F_GIs composed of

The designed total molecular weight is about 31265Da, wherein each PEG chain has a molecular weight of about 5000Da, corresponding to n ₁≈n₂≈n₃≈n₄≈n₅≈n₆≈114。

The preparation process is as follows:

step a: tetrahydrofuran (500mL), a hexahydroxy small-molecule initiator S2-3(1.28g, 2.5mmol) and diphenylmethyl potassium (6.0mmol) were added sequentially to a water-free and oxygen-free closed reaction kettle.

Step c: an excess of methanol, a proton source, was added to give a six-arm polyethylene glycol H1-2 with a carbon atom as a branching center. The hydrogen spectrum data of H1-2 is as follows:¹H NMR(CDCl₃)δ(ppm):3.42-3.76(-OCH₂CH₂OH,24H；>NCH₂CH₂O-,18H；-OCH₂CH₂O-)。

step d: in a dry clean 1L round bottom flask, dry hexa-armed polyethylene glycol H1-2(30.51g,1.0mmol) was added dissolved in DMF (300mL), then 5-azidopentanoic acid (4.29g,30.0mmol), N' -diisopropylcarbodiimide DIC (3.79g,30.0mmol) and DMAP (3.67g,30.0mmol) were added to the previous solution, the reaction was stirred at room temperature for 48 hours, and after completion of the reaction, dialysis was performed for 72 hours to finally obtain hexa-armed polyethylene glycol azide derivative E2-1(24.39g, 78% yield). The hydrogen spectrum data of E2-1 is as follows:¹H NMR(CDCl₃)δ(ppm):1.64-1.86(-CH₂CH₂CH₂CH₂N₃,24H),2.38-2.43(-CH₂CH₂CH₂CH₂N₃,12H),3.27-3.33(-CH₂N₃,12H),3.42-3.76(-OCH₂CH₂O-),4.10-4.30(-OCH₂CH₂OC(O)-,12H)。

GPC measurement of six-arm polyethylene glycol azide derivative E2-1 to determine M_n≈31.3kDa,PDI＝1.03。

Example 7: preparation of six-arm polyethylene glycol ethylamine derivative E3-1 with silicon atom as branched center

Corresponding to the general formula (1), wherein A ₀Is composed of

A₁Is composed of

L_A0A1Is an ether bond-O-, L₂is-CH₂-，F_GIs composed of

The designed total molecular weight is about 21748Da, where each PEG chain has a molecular weight of about 3500Da, corresponding to n₁≈n₂≈n₃≈n₄≈n₅≈n₆≈80。

The preparation process is as follows:

step a: adding 400mL of tetrahydrofuran, a compound S3-1(0.94g,10.0mmol) and excessive diphenyl methyl potassium (75.0mmol) into an anhydrous and oxygen-free closed reaction kettle, then adding a compound S1-2(29.28g,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 hexahydroxy micromolecule initiator S3-4 is obtained through concentration, washing and column chromatography purification. The hydrogen spectrum data of the hexahydroxy small molecule S3-4 are as follows:¹H NMR(CDCl₃)δ(ppm):0.05(-SiCH₃,3H),3.45-3.55(-CH(CH₂OH)₂,3H),3.62-3.78(-CH(CH₂OH)₂,12H)。

step b: tetrahydrofuran (500mL), a hexahydroxy small-molecule initiator S3-4(0.79g,2.5mmol) and diphenylmethyl potassium (6.0mmol) were added sequentially to a water-free and oxygen-free closed reaction kettle.

Step c: adding calculated amount of ethylene oxide, gradually raising the temperature to 60 ℃, and reacting for 48 hours.

Step d: adding excessive proton source methanol to obtain six-arm polyethylene glycol H1-3 with a silicon atom branching center; the hydrogen spectrum data of H1-3 is as follows: ¹H NMR(CDCl₃)δ(ppm):3.33-3.52(-CH(CH₂O-)₂,12H),3.42-3.76(-OCH₂CH₂OH,24H；-OCH₂CH₂O-)。

Step e: in a dry clean 1L round bottom flask, hexa-armed polyethylene glycol H1-3(42.63g,2.0mmol), excess aminoFmoc protected β -alanine (7.47g,24.0mmol) and the solvent dichloromethane (300mL) were added, DMAP (0.04g,0.3mmol) was added, DCC (4.93g,24.0mmol) dissolved in 100mL dichloromethane was added dropwise to the reaction under ice bath conditions,after the 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 aminoFmoc-protected hexa-armed polyethylene glycol ethylamine derivative S3-5(35.09g, yield 76%). The hydrogen spectrum data of S3-5 is as follows:¹H NMR(CDCl₃)δ(ppm):2.45-2.55(-C(O)CH₂CH₂NH-,12H),3.42-3.76(-OCH₂CH₂O-；-C(O)CH₂CH₂NH-,12H),4.08-4.34(Fmoc-9H,6H；-OCH₂CH₂OC(O)-,12H；Fmoc-CH₂-,12H),7.20-7.80(Fmoc-Ar-H,48H)。

step f: removing Fmoc protecting group, treating S3-5(23.08g,1.0mmol) with 20% piperidine/DMF solution, removing Fmoc protection to obtain naked amino, removing solvent by rotary evaporation, dissolving with dichloromethane, precipitating with anhydrous ether, and recrystallizing with isopropanol to obtain hexa-arm polyethylene glycol ethylamine derivative E3-1(21.75 g). The hydrogen spectrum data of E3-1 showed that the characteristic peak of Fmoc disappeared,¹H NMR(CDCl₃)δ(ppm):2.97-3.16(-C(O)CH₂CH₂NH₂,12H)。

GPC measurement of six-arm polyethylene glycol ethylamine derivative E3-1 to determine M_n≈21.7kDa,PDI＝1.03。

Example 8: preparation of six-arm polyethylene glycol propionaldehyde derivative E4-1 with cyclohexane branching center

Corresponding to the general formula (1), wherein A ₀Is composed of

A₁Is composed of

L_A0A1Is an ether bond-O-, L₂is-CH₂-，F_GIs composed of

The designed total molecular weight is about 37291Da, wherein the molecular weight of each PEG chain is about 6000Da, corresponding to n₁≈n₂≈n₃≈n₄≈n₅≈n₆≈136。

The preparation process is as follows:

step a: tetrahydrofuran (500mL), a hexahydroxy small-molecule initiator S4-2(0.89g,2.5mmol) and diphenylmethyl potassium (6.0mmol) were added sequentially to a water-free and oxygen-free closed reaction kettle.

Step c: excess proton source methanol was added to give six-armed polyethylene glycol H1-4 as the cyclohexane branching center. The hydrogen spectrum data of the six-arm polyethylene glycol H1-4 at the cyclohexane branching center are as follows:¹H NMR(CDCl₃)δ(ppm):1.23-2.11(>CHCH₂CH<,6H),3.40-3.78(>CHOCH(CH₂O-)₂,6H；-OCH(CH₂O-)₂,12H；-OCH₂CH₂OH；24H；-OCH₂CH₂O-)。

step d: in a dry clean 1L round bottom flask, dry H1-4(36.35g, 1.0mmol) was dissolved in DMF (200mL) and stirred well before

(S4-3,7.45g,30.0mmol) and DMAP (3.67g,30.0mmol) were added to the above solution, and stirred for 20 minutes. DIC (3.79g, 30.0mmol) dissolved in DMF was then added to the previous solution and stirred overnight. After the reaction was completed, acetone was used for precipitation, and after dissolving in water, dialysis was performed for 24 hours to obtain hexa-arm polyethylene glycol acetal derivative S4-4(30.19g, yield 80%).

Step e: deacetalization of S4-4 was carried out by dissolving S4-4 in dichloromethane in a dry clean 1000mL round-bottomed flask and adding dropwise acetic acid solution to the solution to a pH of 3-4 and stirring for 21 h at room temperatureThen (c) is performed. After the reaction is finished, freeze-drying overnight, and purifying to obtain the final product, namely the hexa-arm polyethylene glycol propionaldehyde derivative E4-1. The hydrogen spectrum data of E4-1 is as follows:¹H NMR(CDCl₃)δ(ppm):2.60-2.78(-C(O)CH₂CH₂C(O)-,24H；-OCH₂CH₂CHO,12H),3.40-3.78(-OCH₂CH₂O-),4.10-4.30(-OCH₂CH₂OC(O)-,12H),4.50-4.60(-OCH₂CH₂CHO,12H),9.77(-CH₂CH₂CHO,6H)。

GPC measurement of six-arm polyethylene glycol propionaldehyde derivative E4-1 to determine M_n≈37.3kDa,PDI＝1.04。

Example 9: preparation of nitrogen atom branched center six-arm polyethylene glycol propionic acid derivative E5-1

Corresponding to the general formula (1), wherein A₀Is composed of

A₁Is composed of

L_A0A1Containing an ether bond of-CH₂CH₂OCH₂-，L₂is-OCH₂CH₂-，F_GIs composed of

The designed total molecular weight is about 16104Da, wherein each PEG chain has a molecular weight of about 2500Da and a degree of polymerization n corresponding to each PEG chain₁-1≈n₂-1≈n₃-1≈n₄-1≈n₅-1≈n₆-1≈57。

The preparation process is as follows:

step a: 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 S5-1(32.60g,75.0mmol), and reacting at 30 ℃ for 12 hours; and after the reaction is finished, opening the reaction kettle, washing and concentrating the reaction solution, dissolving the reaction solution by using methanol, adding 1M hydrochloric acid until the pH value is 3.5, reacting for 4 hours, and concentrating, washing and purifying by column chromatography to obtain the hexahydroxy micromolecule initiator S5-2. The hydrogen spectrum data of the hexahydroxy small molecule S5-2 are as follows: ¹H NMR(CDCl₃)δ(ppm):0.45(-SiCH₃,9H),2.60-2.70(>NCH₂CH₂O-,6H),3.02-3.17(-SiCH₂O-,6H；-Si(CH₂OH)₂,12H),3.48-3.68(>NCH₂CH₂O-,6H)。

Step b: tetrahydrofuran (500mL), a hexahydroxy small-molecule initiator S5-2(1.25g,2.5mmol) and diphenylmethyl potassium (6.0mmol) were added sequentially to a water-free and oxygen-free closed reaction kettle.

Step d: an excess of methanol, a proton source, was added to give a six-armed polyethylene glycol H1-5 with a nitrogen atom as a branching center. The hydrogen spectrum data of H1-5 is as follows:¹H NMR(CDCl₃)δ(ppm):3.02-3.12(-Si(CH₂OCH₂-)₃,18H),3.55-3.75(-OCH₂CH₂OH,24H；-OCH₂CH₂O-)。

step e: six-armed polyethylene glycol H1-5(15.50g,1.0mmol) was dissolved in toluene (300mL), and excess succinic anhydride (3.0g,30.0mmol) was added at 50 ℃ for 12 hours. After the reaction was completed, the reaction vessel was opened, the solvent was concentrated, and the product was precipitated in anhydrous ether at 0 ℃, filtered, dried, and purified with silica gel column to obtain hexa-armed polyethylene glycol propionic acid derivative E5-1(13.37g, yield 83%). The hydrogen spectrum data of E5-1 is as follows:¹H NMR(CDCl₃)δ(ppm):2.58-2.64(-C(＝O)CH₂CH₂COOH,24H),3.02-3.12(-Si(CH₂OCH₂-)₃,18H),3.45-3.85(-OCH₂CH₂OC(O)-,12H；-OCH₂CH₂O-),4.10-4.25(-OCH₂CH₂OC(O)-,12H)。

GPC measurement of six-arm polyethylene glycol propionic acid derivative E5-1 to determine M_n≈16.1kDa,PDI＝1.02。

Example 10: preparation of nitrogen branched center six-arm polyethylene glycol lipoic acid derivative E6-1

Corresponding to the general formula (1), wherein A₀Is composed of

A₁Is composed of

L_A0A1Containing an ether bond of-CH₂CH₂OCH₂CH₂-，L₂is-CH₂-，F_GIs composed of

The designed total molecular weight is about 4490Da, wherein each PEG chain has a molecular weight of 500Da, corresponding to n ₁＝n₂＝n₃＝n₄＝n₅＝n₆＝n＝11。

The preparation process is as follows:

step a: 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 S6-1(31.39g,75.0mmol), and reacting at 30 ℃ for 12 hours; opening the reaction kettle, washing, concentrating, dissolving with methanol, adding 1M hydrochloric acid until pH is 3.5, reacting for 4 hr, and concentratingWashing and column chromatography to obtain the hexahydroxy micromolecular initiator S6-2. The hydrogen spectrum data of S6-2 is as follows:¹HNMR(CDCl₃)δ(ppm):1.79(-CH(CH₂OH)₂,3H),2.60-2.70(>NCH₂CH₂O-,6H),3.48-3.68(>NCH₂CH₂O-,6H),3.70-3.84(-CH(CH₂OH)₂,12H)。

step b: tetrahydrofuran (500mL), a hexahydroxy small-molecule initiator S6-2(1.14g,2.5mmol) and diphenylmethyl potassium (6.0mmol) were added sequentially to a water-free and oxygen-free closed reaction kettle.

Step d: an excess of methanol, a proton source, was added to give a six-armed polyethylene glycol H1-6 with a nitrogen atom as a branching center. The hydrogen spectrum data of H1-6 is as follows:¹H NMR(CDCl₃)δ(ppm):3.40-3.78(-OCH(CH₂O-)₂,12H；-OCH₂CH₂OH)。

step e: in a dry clean 1L round bottom flask, hexa-armed polyethylene glycol H1-6(6.72g,2.0mmol), excess lipoic acid compound S6-3(4.95g,24.0mmol) and solvent dichloromethane (100mL) were added, DMAP (0.04g,0.3mmol) was added, DCC (4.93g,24.0mmol) dissolved in 100mL dichloromethane was added dropwise to the reaction solution under ice bath, and after the addition was completed, 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 hexa-armed polyethylene glycol lipoic acid derivative E6-1(7.0g, yield 78%). The hydrogen spectrum data of E6-1 is as follows: ¹H NMR(CDCl₃)δ(ppm):1.40-1.48(-C(O)CH₂CH₂CH₂CH₂CH<,12H),1.60-1.68(-C(O)CH₂CH₂CH₂CH₂CH<,24H),1.85-1.91(-SCH₂CH₂-,12H),2.25-2.31(-C(O)CH₂CH₂CH₂CH₂CH<,12H),3.07-3.13(-SCH₂CH₂-,12H),3.51-3.61(-SCHCH₂-,6H)。

GPC measurement of six-arm polyethylene glycol lipoic acid derivative E6-1 to determine M_n≈4.5kDa,PDI＝1.01。

Example 11: preparation of nitrogen atom branched center six-arm polyethylene glycol hydroxyl derivative E7-1

Corresponding to the general formula (1), wherein A₀Is composed of

A₁Is composed of

L_A0A1Containing an ether bond of-CH₂CH₂O-，L₂Is absent, F_GIs composed of

The designed total molecular weight is about 7008Da, wherein each PEG chain has a molecular weight of about 1000Da corresponding to n₁≈n₂≈n₃≈n₄≈n₅≈n₆≈23。

The preparation process is as follows:

step a: 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 S7-1(33.50g,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 hexahydroxy micromolecule initiator S7-2 is obtained through concentration, washing and column chromatography purification. The hydrogen spectrum data of the hexahydroxy small molecule S7-2 are as follows:¹H NMR(CDCl₃)δ(ppm):1.60-2.11(-CH(OH)CH₂CH(OH)-,6H),2.60-2.70(>NCH₂CH₂O-,6H),3.48-3.68(>NCH₂CH₂O-,6H；>CHOCH₂CH₂-,3H),3.71-3.83(-CH(OH)CH₂CH(OH)-,6H)。

step b: tetrahydrofuran (500mL), a hexahydroxy small-molecule initiator S7-2(1.23g,2.5mmol) and diphenylmethyl potassium (6.0mmol) were added sequentially to a water-free and oxygen-free closed reaction kettle.

Step d: an excess of methanol, a proton source, was added to give a six-armed polyethylene glycol H1-7 with a nitrogen atom as a branching center. The hydrogen spectrum data of H1-7 is as follows:¹H NMR(CDCl₃)δ(ppm):3.49-3.75(>CHOCH₂CH₂-,9H；-OCH₂CH₂OH,24H)。

step e: in a dry clean 1L round bottom flask, hexa-armed polyethylene glycol H1-7(12.98g,2.0mmol), excess hydroxy EE protected 4-hydroxybutyric acid (S7-3,4.23g,24.0mmol) and solvent dichloromethane (100mL) were added, DMAP (0.04g,0.3mmol) was added, DCC (4.93g,24.0mmol) dissolved in 100mL dichloromethane was added dropwise to the reaction solution under ice bath, and after the addition was completed, the reaction was carried out at room temperature for 16H. After completion of the reaction, insoluble matter was removed by filtration, concentrated and purified by column chromatography to give hydroxy EE-protected six-armed polyethylene glycol derivative S7-4(12.05g, yield 81%).

Step f: removing an EE protecting group from S7-4, adding the hexa-armed polyethylene glycol derivative S7-4(7.44g,1.0mmol) protected by the hydroxy EE prepared in the previous step into a dry and clean container, dissolving the mixture by using methanol, adding 1M hydrochloric acid into the mixture until the pH value is 3.5, and reacting the mixture for 4 hours to obtain the hexa-armed polyethylene glycol E7-1(7.01g) with exposed hydroxy. The hydrogen spectrum data of E7-1 is as follows:¹H NMR(CDCl₃)δ(ppm):1.45-1.53(-CH₂CH₂CH₂OH,12H),2.33-2.38(-OC(O)CH₂CH₂CH₂-,12H),3.40-3.78(-OCH₂CH₂O-；-CH₂CH₂CH₂OH,12H),4.10-4.30(-C(O)OCH₂CH₂O-,12H)。

GPC measurement of six-arm polyethylene glycol hydroxy derivative E7-1 to determine M_n≈7.0kDa,PDI＝1.02。

Example 12: preparation of benzyl triol branching center six-arm polyethylene glycol maleimide derivative E8-1

Corresponding to the general formula (1), wherein A₀Is composed of

A₁Is composed of

L_A0A1Containing an ether bond of-CH₂O-，L₂is-CH₂-，F_GIs composed of

The designed total molecular weight is about 19297Da, wherein each PEG chain has a molecular weight of about 3000Da, corresponding to n₁≈n₂≈n₃≈n₄≈n₅≈n₆≈68。

The preparation process is as follows:

step a: adding 400mL of tetrahydrofuran, a compound S8-1(1.68g,10.0mmol) and excessive diphenyl methyl potassium (75.0mmol) into an anhydrous and oxygen-free closed reaction kettle, then adding a compound S1-2(29.28g,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 hexahydroxy micromolecule initiator S8-2 is obtained through concentration, washing and column chromatography purification. The hydrogen spectrum data of S8-2 is as follows:¹H NMR(CDCl₃)δ(ppm):3.45-3.55(-CH(CH₂OH)₂,3H),3.62-3.78(-CH(CH₂OH)₂,12H),4.47(ArCH₂O-,6H),7.24(Ar-H,3H)。

step b: tetrahydrofuran (500mL), a hexahydroxy small-molecule initiator S8-2(0.98g,2.5mmol) and diphenylmethyl potassium (6.0mmol) were added sequentially to a water-free and oxygen-free closed reaction kettle.

Step d: excess of proton source methanol was added to afford six-armed polyethylene glycol H1-8. The hydrogen spectrum data of H1-8 is as follows: ¹H NMR(CDCl₃)δ(ppm):3.33-3.52(-CH(CH₂O-)₂,12H),3.55-3.75(-OCH₂CH₂OH,24H；-OCH₂CH₂O-)。

Step e: in a dry clean 1L round bottom flask, hexa-armed polyethylene glycol H1-8(36.78g,2.0mmol), excess maleimide propionic acid (4.06g,24.0mmol) and solvent dichloromethane (100mL) were added, DMAP (0.04g,0.3mmol) was added under ice bath conditions, DCC (4.93g,24.0mmol) dissolved in 100mL 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 completion of the reaction, insoluble matter was removed by filtration, concentrated and purified by column chromatography to give six-arm polyethylene glycol maleimide derivative E8-1(30.88g, yield 80%). The hydrogen spectrum data of E8-1 is as follows:¹H NMR(CDCl₃)δ(ppm):2.61-2.68(-C(O)CH₂CH₂N<,12H),3.28-3.57(-OCH₂CH₂O-；-OCH₂CH₂OC(O)-,12H),3.63-3.87(-C(O)CH₂CH₂N<,12H),4.10-4.30(-OCH₂CH₂OC(O)-,12H),6.71(-CH＝CH-,12H)。

GPC measurement of six-arm polyethylene glycol maleimide derivative E8-1 to determine M_n≈19.3kDa,PDI＝1.03。

Example 13: preparation of six-arm polyethylene glycol propylene derivative E9-1 with carbon atom branching center

Corresponding to the general formula (1), wherein A₀Is composed of

A₁Is composed of

L_A0A1Containing an ether bond of-CH₂OCH₂-，L₂is-CH₂-，F_GIs composed of

The designed total molecular weight is about 60811Da, wherein each PEG chain has a molecular weight of about 10000Da, corresponding to n₁≈n₂≈n₃≈n₄≈n₅≈n₆≈227。

The preparation process is as follows:

step a: adding 400mL of tetrahydrofuran, a compound S4-1(1.20g,10.0mmol) and excessive diphenyl methyl potassium (75.0mmol) into an anhydrous and oxygen-free closed reaction kettle, and then adding a compound S9-1(36.20g,75.0mmol) into the reaction kettle, wherein the reaction temperature is 30 ℃, and the reaction time is 12 hours; and opening the reaction kettle, washing, concentrating, dissolving with methanol, adding 1M hydrochloric acid until the pH value is 3.5, reacting for 4 hours, concentrating, washing and performing column chromatography to obtain the hexahydroxy micromolecule initiator S9-2. The hydrogen spectrum data of S9-2 is as follows: ¹H NMR(CDCl₃)δ(ppm):0.81(-CCH₃,3H),3.61(-CCH₂O-,6H),4.41(Ar-CH₂OH,12H),4.47(ArCH₂O-,6H),7.24(Ar-H,9H)。

Step b: tetrahydrofuran (500mL), a hexahydroxy small-molecule initiator S9-2(1.43g,2.5mmol) and diphenylmethyl potassium (6.0mmol) were added sequentially to a closed anhydrous and oxygen-free reaction vessel.

Step d: adding an excess of proton sourceMethanol to obtain six-arm polyethylene glycol H1-9. The hydrogen spectrum data of the six-arm polyethylene glycol H1-9 are as follows:¹H NMR(CDCl₃)δ(ppm):3.42-3.75(-OCH₂CH₂OH,24H；-OCH₂CH₂O-),4.47(ArCH₂O-,18H)。

step e: in a dry clean 1L round bottom flask, six-armed polyethylene glycol H1-9(60.57g,1.0mmol) was added, excess diphenylmethyl potassium (10.0mmol) was added, followed by excess propenyl chloride (0.77g,10.0mmol), reaction temperature was 30 ℃ and reaction time was 12 hours; opening the reaction kettle, concentrating the solvent, precipitating in anhydrous ether at 0 deg.C, filtering, and drying to obtain hexa-arm polyethylene glycol propenyl ether derivative E9-1(24.32 g). The hydrogen spectrum data of E9-1 is as follows:¹H NMR(CDCl₃)δ(ppm):3.42-3.75(-OCH₂CH₂O-；-OCH₂CH₂OC(O)-,12H；-OCH₂CH＝CH₂,12H),5.15-5.32(-OCH₂CH＝CH₂,12H),5.82-5.98(-OCH₂CH＝CH₂,6H)。

GPC measurement of six-arm polyethylene glycol propenyl ether derivative E9-1 to determine M_n≈60.8kDa,PDI＝1.05。

Example 14: preparation of six-arm polyethylene glycol acrylate derivative E10-1

Corresponding to the general formula (1), wherein A₀Is composed of

A₁Is composed of

L_A0A1Containing an ether bond of-OCH₂-，L₂is-CH₂-，F_GIs composed of

The designed total molecular weight is about 25429Da, wherein each PEG chain has a molecular weight of about 4000Da corresponding to n ₁≈n₂≈n₃≈n₄≈n₅≈n₆≈91。

The preparation process is as follows:

step a: adding 400mL of tetrahydrofuran, a compound S5-1(1.32g,10.0mmol) and excessive diphenyl methyl potassium (75.0mmol) into an anhydrous and oxygen-free closed reaction kettle, and then adding a compound S9-1(36.20g,75.0mmol) into the reaction kettle, 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 hexahydroxy micromolecule initiator S10-1 is obtained through concentration, washing and column chromatography purification. The hydrogen spectrum data of S10-1 is as follows:¹H NMR(CDCl₃)δ(ppm):1.23-2.11(>CHCH₂CH<,6H),3.40-3.55(>CHOCH₂-,3H),4.41(Ar-CH₂OH,12H),4.51(Ar-CH₂O-,6H),7.24(Ar-H,9H)。

step b: tetrahydrofuran (500mL), a hexahydroxy small-molecule initiator S10-1(1.46g,2.5mmol) and diphenylmethyl potassium (6.0mmol) were added sequentially to a water-free and oxygen-free closed reaction kettle.

Step d: excess proton source methanol was added to give six-armed polyethylene glycol H1-10, the cyclohexane branching center. The hydrogen spectrum data of H1-10 are as follows:¹H NMR(CDCl₃)δ(ppm)：3.42-3.75(-OCH₂CH₂OH,24H；-OCH₂CH₂O-),4.47(ArCH₂OCH₂-,12H)。

step e: in a dry clean 1L round bottom flask, hexa-armed polyethylene glycol H1-10(24.58g,1.0mmol) and 4,4' -methylenebis (2, 6-di-tert-butylphenol) (0.9 wt%) were dissolved in anhydrous DMF (150mL) and excess isocyanate ethyl acrylate S10-2(2.54g,18.0 m) mol) and the catalyst dibutyltin dilaurate (DBTDL,0.1 wt%) were added to the above solution, and the reaction was stirred at 40 ℃ for 24 to 36 hours. After the reaction is finished, the reaction kettle is opened, the solvent is concentrated, and then the product is precipitated twice in anhydrous ether at the temperature of 0 ℃, filtered and dried, so that the hexa-arm polyethylene glycol acrylate derivative E10-1(21.62g, the yield is 85%) is obtained. The hydrogen spectrum data of E10-1 is as follows:¹H NMR(DMSO-d₆)δ(ppm):3.50-3.75(-OCH₂CH₂O-；-OCH₂CH₂OC(O)-,12H；-OC(O)NHCH₂-,12H),3.78-4.18(-OCH₂CH₂OC(O)-,12H；-NHCH₂CH₂OC(O)-,12H),4.95-5.23(-CH＝CH₂,12H),5.67-5.96(-CH＝CH₂,6H)。

GPC measurement of six-arm polyethylene glycol acrylate derivative E10-1 to determine M_n≈25.4kDa,PDI＝1.03。

Example 15: preparation of six-arm polyethylene glycol derivative E11-1 with branch chain end as branch structure

Corresponding to the general formula (1), wherein A₀Is composed of

A₁Is composed of

L_A0A1Containing an ether bond of-CH₂CH₂OCH₂-，L₂is-CH₂-，F_GIs composed of

(g＝1,k＝3,L₀＝-COCH₂CH₂CONH-,

q＝0,Z₂Is absent, q₁＝0,Z₁Is absent, R₀₁＝OPG₄,PG₄TBS). The designed total molecular weight is about 18738Da, wherein each PEG chain has a molecular weight of about 2500Da, corresponding to n₁≈n₂≈n₃≈n₄≈n₅≈n₆≈56。

The preparation process is as follows:

3.20g of six-arm polyethylene glycol E2-1 with six carboxyl groups (toluene is removed by azeotropic dehydration), 20mL of triethylamine and 55.68g of compound S11-1 are added into a dry and clean 1L round bottom flask, solvent dichloromethane (300mL) is added under nitrogen protection, the mixture is stirred until the mixture is dissolved, 29.61g of Dicyclohexylcarbodiimide (DCC) is added, reaction is carried out for 24 hours at room temperature, insoluble substances are removed by filtration, concentration, isopropanol recrystallization and column chromatography purification are carried out, and the six-arm polyethylene glycol hydroxyl derivative E11-1 is obtained. The hydrogen spectrum data of E11-1 is as follows: ¹H NMR(CDCl₃)δ(ppm):0.21(-Si(CH₃)₂,108H),0.98(-SiC(CH₃)₃,162H),2.80-3.00(-C(O)CH₂CH₂C(O)-,24H),3.40-3.80(-OCH₂CH₂O-),3.90-4.20(-C(O)NHC(CH₂O-)₃,36H)。

GPC measurement of six-arm polyethylene glycol derivative E11-1 to determine M_n≈18.7kDa,PDI＝1.03。

Example 16: preparation of hexa-arm polyethylene glycol derivative E12-1 with comb-shaped branched chain end

Corresponding to the general formula (1), wherein A₀Is a nitrogen atom

A₁Is composed of

The designed total molecular weight is about 39275Da, wherein each PEG chain has a molecular weight of about 1500Da, corresponding to n₁≈n₂≈n₃≈n₄≈n₅≈n₆≈n₇≈34。

The preparation process is as follows:

step a: into a water-free and oxygen-free closed reaction vessel were added 200mL of tetrahydrofuran, 6-hydroxy six-armed polyethylene glycol H1-1(18.74g,2.0mmol), and diphenylmethyl potassium (9.6mmol) in that order.

Step b: a calculated amount of S12-1(Ethoxy ethyl glycosyl ether,2400mmol) was added, and the temperature was gradually raised to 60 ℃ for 48 hours.

Step c: adding excessive diphenyl methyl potassium (24.0mmol), adding excessive methyl iodide (60.0mmol), and reacting at 30 deg.c for 12 hr; opening the reaction kettle, concentrating the solvent, precipitating in anhydrous ether at 0 ℃, filtering, and drying to obtain the hexa-arm polyethylene glycol derivative E12-1; the hydrogen spectrum data of E12-1 is as follows:¹H NMR(CDCl₃)δ(ppm):1.22(-OCH₂CH₃),1.36(-OCH(O)CH₃),3.35(-OCH₃),3.40-3.80(-OCH₂CH₂O-,-OCH(CH₂O-)₂,OCH₂CH₃),4.75(-OCHCH₃(OCH₂))。

GPC measurement of six-arm polyethylene glycol derivative E12-1 to determine M_n≈39.3kDa,PDI＝1.04。

Example 17: preparation of six-arm polyethylene glycol hydroxyl derivative E13-1 with hyperbranched structure at branch chain end

Corresponding to the general formula (1), wherein A₀Is a nitrogen atom

A₁Is composed of

L_A0A1Containing an ether bond of-CH₂CH₂O-，L₂is-CH₂-. The designed total molecular weight is about 19016Da, wherein each PEG chain has a molecular weight of about 1500Da, corresponding to n₁≈n₂≈n₃≈n₄≈n₅≈n₆≈34。

The preparation process is as follows:

Step b: a calculated amount of compound S13-1(2400mmol) 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 six-arm polyethylene glycol hydroxy derivative E13-1; the hydrogen spectrum data of compound E13-1 are as follows:¹H NMR(CDCl₃)δ(ppm):3.40-3.80(-OCH₂CH₂O-；-OCH(CH₂O-)₂；-OCH₂CH(O)CH(O)-),3.85-4.40(-OCH₂CH(O)CH(O)-)。

derivatization of hydroxyl group of polyethylene glycol with six armsObject E13-1 was subjected to GPC measurement to determine M_n≈19.0kDa,PDI＝1.03。

Example 18: preparation of six-arm polyethylene glycol hydroxyl derivative E14-1 with hyperbranched structure at branch chain end

Corresponding to the general formula (1), wherein A₀Is a nitrogen atom

A₁Is composed of

L_A0A1Containing an ether bond of-CH₂CH₂O-，L₂is-CH₂-. The designed total molecular weight is about 16483Da, wherein each PEG chain has a molecular weight of about 1500Da corresponding to n₁≈n₂≈n₃≈n₄≈n₅≈n₆≈34。

The preparation process is as follows:

step a: 200mL of tetrahydrofuran, 6-hydroxy six-armed polyethylene glycol H1-1(18.74g,2.0mmol), and diphenylmethyl potassium (9.6mmol) were added sequentially.

Step b: a calculated amount of glycidol S14-1(2400mmol) 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 six-arm polyethylene glycol hydroxy derivative E14-1; the hydrogen spectrum data of E14-1 is as follows:¹H NMR(CDCl₃)δ(ppm):3.40-3.85(-CH₂CH₂O-,-OCH(CH₂O-)₂)。

GPC measurement of the six-arm polyethylene glycol hydroxy derivative E14-1 was conducted to confirmM_n≈16.5kDa,PDI＝1.03。

Example 19: preparation of six-arm polyethylene glycol hydroxyl derivative E15-1 with branch chain end in dendritic structure

Corresponding to the general formula (1), wherein A₀Is a nitrogen atom

A₁Is composed of

L_A0A1Containing an ether bond of-CH₂CH₂O-，L₂is-CH₂-. The designed total molecular weight is about 12482Da, wherein each PEG chain has a molecular weight of about 1500Da, corresponding to n₁≈n₂≈n₃≈n₄≈n₅≈n₆≈34。

The preparation process is as follows:

step a: adding 200mL of tetrahydrofuran, hexa-armed polyethylene glycol H1-1(18.74g,2.0mmol) and excessive diphenyl methyl potassium (96.0mmol) into a water-free and oxygen-free closed reaction kettle, and then adding excessive compound S15-1(240mmol), wherein the reaction temperature is 30 ℃, and the reaction time is 12 hours; opening the reaction kettle, concentrating the solvent, precipitating in anhydrous ether at 0 ℃, filtering and drying to obtain a six-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: ¹H NMR(CDCl₃)δ(ppm):0.21(-Si(CH₃)₂),0.98(-SiC(CH₃)₃),2.90-3.10(-OCHCH₂OSi-),3.40-3.80(-CH₂CH₂O-,-OCH(CH₂O-)₂),3.80-4.10(-OCHCH₂OSi-)。

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 hydroxyl-exposed hexa-arm polyethylene glycol intermediate S15-3.

Step c: and (c) repeating the steps a and b twice to obtain the dendritic six-arm polyethylene glycol hydroxyl derivative E15-1 with naked hydroxyl.

GPC measurement of six-arm polyethylene glycol hydroxy derivative E15-1 to determine M_n≈12.5kDa,PDI＝1.03。

Example 20: preparation of six-arm polyethylene glycol hydroxyl derivative E16-1 with branched structure at branch chain end

Corresponding to the general formula (1), wherein A₀Is a nitrogen atom

A₁Is composed of

The designed total molecular weight is about 10242Da, wherein each PEG chain has a molecular weight of about 1500Da, corresponding to n₁≈n₂≈n₃≈n₄≈n₅≈n₆≈34。

The preparation process is as follows:

step a: in a dry clean 1L round bottom flask, hexa-armed polyethylene glycol H1-1(18.74g,2.0mmol), excess S16-1(7.38g,24.0mmol) and solvent dichloromethane (100mL) were added, DMAP (0.04g,0.3mmol) was added under ice bath, DCC (4.93g,24.0mmol) dissolved in 100mL dichloromethane was added dropwise to the reaction, and DCC (4.93g,24.0mmol) was added dropwiseAfter the reaction is finished, the reaction is carried out for 16h at room temperature. After completion of the reaction, insoluble matter was removed by filtration, concentrated and purified by column chromatography to give hydroxy EE-protected six-armed polyethylene glycol intermediate S16-2(16.88g, yield 76%). The hydrogen spectrum data of S16-2 is as follows: ¹H NMR(CDCl₃)δ(ppm):0.21(-Si(CH₃)₂,96H),0.98(-SiC(CH₃)₃,108H),3.40-3.46(>NCH₂C(O)O-,12H)。

Step b: and (3) removing an EE protecting group, dissolving S16-2 in methanol, adding 1M hydrochloric acid until the pH value is 3.5, reacting for 4 hours, concentrating, washing and carrying out column chromatography to obtain the six-arm polyethylene glycol hydroxyl derivative E16-1. The structure of E16-1 was determined by NMR. The hydrogen spectrum data of E16-1 is as follows:¹H NMR(CDCl₃)δ(ppm):3.76-3.80(-N(CH₂CH₂OH)₂,24H)。

GPC measurement of six-arm polyethylene glycol hydroxy derivative E16-1 to determine M_n≈10.2kDa,PDI＝1.02。

Example 21: preparation of hexa-arm polyethylene glycol amine derivative E17-1 with branch chain end in dendritic structure

Corresponding to the general formula (1), wherein A₀Is a nitrogen atom

A₁Is composed of

Designed overall molecular weight of about11678Da, wherein each PEG chain has a molecular weight of about 1500Da, corresponding to n₁≈n₂≈n₃≈n₄≈n₅≈n₆≈34。

The preparation process is as follows:

step a: in a dry clean 1L round bottom flask, hexa-armed polyethylene glycol H1-1(18.74g,2.0mmol), excess trilysine S17-1(30.66g,24.0mmol) and solvent dichloromethane (200mL) were added, DMAP (0.04g,0.3mmol) was added under ice bath, DCC (4.93g,24.0mmol) dissolved in 100mL dichloromethane was added dropwise to the reaction mixture, and after the addition was completed, the reaction was carried out at room temperature for 16H. After completion of the reaction, insoluble matter was removed by filtration, concentrated and purified by column chromatography to give aminofmoc-protected six-armed polyethylene glycol intermediate S17-2(19.52g, yield 75%).

Step b: removing Fmoc protecting groups, treating S17-2 with 20% piperidine/DMF solution, removing Fmoc protection to obtain naked amino, removing the solvent by rotary evaporation, dissolving with dichloromethane, precipitating with anhydrous ether, and recrystallizing with isopropanol to obtain the hexa-arm polyethylene glycol amine derivative E17-1. The structure of E17-1 was determined by NMR.

GPC measurement of six-arm polyethylene glycol amine derivative E17-1 to determine M_n≈11.7kDa,PDI＝1.02。

Example 22: preparation of six-arm polyethylene glycol azide derivative E18-1 (coupling method)

Corresponding to the general formula (1), wherein A₀Is composed of

A₁Is composed of

L_A0A1Containing an amide bond as-CH₂C(O)NH-，L₂is-CH₂-，F_Gis-CH₂CH₂N₃. The designed total molecular weight is about 6897Da, wherein each PEG chain has a molecular weight of about 1000Da corresponding to n₁≈n₂≈n₃≈n₄≈n₅≈n₆＝n+1＝23。

The preparation process is as follows:

step a: in a dry clean 250mL round bottom flask, the triacylglyceride-modified acid derivative S18-1(1.45g,3.0mmol) was dissolved in 100mL of dichloromethane, the amine derivative S18-2(1.69g, 7.2mmol) containing two TBS protected hydroxyl groups was added and the reaction stirred at room temperature for 12 hours. After the reaction is finished, filtering to obtain a colorless dichloromethane solution, precipitating in glacial ethyl ether, recrystallizing for 2 times by using ethanol, and purifying by using a column to obtain a hexafunctional small molecular intermediate containing six TBS protected hydroxyl groups; and (3) carrying out deprotection reaction to remove TBS protecting groups, dissolving the obtained hexafunctional micromolecule intermediate containing six TBS protected hydroxyl groups in tetrahydrofuran, adding tetra-tert-butylammonium fluoride (TBAF), reacting overnight, removing TBS protection, and purifying by silica gel column chromatography to obtain hexafunctional micromolecule S18-3(1.02g, yield 83%) containing six naked hydroxyl groups. The hydrogen spectrum data of S18-3 is as follows: ¹H NMR(CDCl₃)δ(ppm):3.26(N(CH₂CONH-)₃,6H),3.90-3.94(-CH(CH₂OH)₂,15H)。

Step b: dissolving hexafunctional micromolecule S18-3(0.41g,1.0mmol) in DMF in an anhydrous and oxygen-free closed reaction kettle, and then adding potassium carbonate (0.46g,3.3mmol) and benzenesulfonyl polyethylene glycol azide derivative dissolved in DMF

(S18-4,1.95g, 1.5mmol), the reaction was stirred at 80 ℃ for 24 hours. After the reaction is finished, cooling the reaction liquid to room temperature, then filtering to remove insoluble substances, concentrating, extracting, combining organic phases, washing with brine, drying with anhydrous sodium sulfate, filtering, concentrating, and purifying by silica gel column chromatography to obtain the nitrogen-branched six-arm polyethylene glycol azide derivative E18-1 (3.7)9g) In that respect The hydrogen spectrum data of E18-1 is as follows:¹H NMR(CDCl₃)δ(ppm):3.30-3.40(-OCH₂CH₂N₃,12H),3.45-3.80(-OCH₂CH₂N₃,12H；-OCH₂CH₂O-)。

GPC measurement of six-arm polyethylene glycol azide derivative E18-1 to determine M_n≈6.9kDa,PDI＝1.02。

Example 23: preparation of hexa-armed polyethylene glycol ethylamine derivative E19-2 (coupling method)

Corresponding to the general formula (1), wherein A₀Is composed of

A₁Is composed of

L_A0A1Containing ester bonds as-CH₂C(O)OCH₂CH₂-，L₂is-OC (O) OCH₂CH₂-，F_Gis-CH²CH₂NH₂. The designed total molecular weight is about 13628Da, wherein each PEG chain has a molecular weight of about 2000Da corresponding to n₁≈n₂≈n₃≈n₄≈n₅≈n₆≈n＝46。

The preparation process is as follows:

step a: in a dry and clean 500mL round-bottom flask, adding a small molecular compound NOTA- (COOH) under the protection of nitrogen₃(S19-1,0.91g, 3.0mmol), compound S19-2(4.76g, 12.6mmol) containing two TBS protected hydroxyl groups and one naked hydroxyl group, 4-dimethylaminopyridine DMAP (0.02g, 0.18mmol), 150mL of dichloromethane, stirred for 30 minutes in ice bath, and then A solution of dicyclohexylcarbodiimide DCC (2.22g, 10.8mmol) dissolved in 20mL of methylene chloride was slowly added dropwise to the mixture, and after completion of the addition, the mixture was returned to room temperature and the reaction was stirred for 15 hours. After the reaction is finished, filtering to remove generated insoluble substances, concentrating, washing with a saturated sodium bicarbonate solution, washing with water to remove residual sodium bicarbonate, extracting, combining organic phases, drying with anhydrous magnesium sulfate, filtering, concentrating, and purifying by silica gel column chromatography to obtain a hexafunctionalized small molecular intermediate containing six TBS protected hydroxyl groups; and (3) carrying out deprotection reaction to remove TBS protecting groups, dissolving the obtained hexafunctional micromolecule intermediate containing six TBS protected hydroxyl groups in tetrahydrofuran, adding tetra-tert-butyl ammonium fluoride (TBAF), reacting overnight, removing TBS protection, and purifying by silica gel column chromatography to obtain hexafunctional micromolecule S19-3(1.57g, yield 75%) containing six naked hydroxyl groups. The hydrogen spectrum data of S19-3 is as follows:¹H NMR(CDCl₃)δ(ppm):2.60-2.93(>NCH₂CH₂N<,12H)；3.10-3.40(>NCH₂C(O)O-,6H),3.58-3.62(>NCH₂CH₂OCO-,6H；-N(CH₂CH₂OH)₂,12H),4.15-4.25(>NCH₂CH₂OCO-,6H)。

step b: in a dry clean 1000mL round bottom flask, amino Fmoc protected polyethylene glycol succinimide carbonate

(S19-4, azeotropic removal of water with toluene, 2492Da, PDI 1.01,7.2mmol) was dissolved in dichloromethane, 4-Dimethylaminopyridine (DMAP) (0.13g,1.1mmol) was added thereto, the mixture was stirred and mixed, hexafunctional small molecule S19-3(0.70g,1.0mmol) was added to the reaction mixture, and the reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction, the reaction mixture was spin-dried, recrystallized from isopropanol, and purified by ion exchange resin to obtain the aminofmoc-protected hexa-armed polyethylene glycol amine derivative E19-1(12.27g, yield 82%). The hydrogen spectrum data of E19-1 is as follows: ¹H NMR(CDCl₃)δ(ppm):3.00-3.20(-OCH₂CH₂NH-,12H),3.45-3.75(-OCH₂CH₂NH-,12H；-OCH₂CH₂O-；-C(O)OCH₂CH₂O-,12H),3.90-4.20(-CH₂OC(O)OCH₂-,24H；Fmoc-9H,6H),4.24-4.28(Fmoc-CH₂-,12H),7.20-7.80(Fmoc-Ar,48H)。

Step c: deprotecting E19-1 containing Fmoc protected amino, treating E19-1 with 20% piperidine/DMF solution, removing the solvent by rotary evaporation, dissolving with dichloromethane, precipitating with anhydrous ether, and recrystallizing with isopropanol to obtain hexa-arm polyethylene glycol ethylamine derivative E19-2 containing six 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%.

GPC measurement of six-arm polyethylene glycol ethylamine derivative E19-2 to determine M_n≈13.6kDa,PDI＝1.03。

Example 24: preparation of six-armed polyethylene glycol TBS protected hydroxy derivative E20-1 (coupling method)

Corresponding to the general formula (1), wherein A₀Is composed of

A₁Is composed of

L_A0A1Is an amide bond-C (O) NH-, L₂is-OC (O) NHCH₂CH₂-，F_Gis-CH₂CH₂OTBS. The designed total molecular weight is about 19856Da, wherein each PEG chain has a molecular weight of about 3000Da, corresponding to n₁≈n₂≈n₃≈n₄≈n₅≈n₆≈n＝68。

The preparation process is as follows:

step a: in a round-bottomed flask, a compound containing an Fmoc-protected amino group and two naked hydroxyl groups is addedCompound S20-1(

0.63g, 2mmol) and nitrogen protection, dissolving with dichloromethane, adding catalyst dibutyltin dilaurate (0.5mL) under nitrogen protection, then slowly dropwise adding polyethylene glycol isocyanate TBSOPEG-NCO (subjected to azeotropic dehydration with toluene, 3kDa, PDI ═ 1.01, 4.8mmol) protected by hydroxy TBS under ice-bath, and stirring at room temperature for reaction for 8 hours after dropwise addition. And after the reaction is finished, adding excessive activated silica gel, filtering, concentrating, recrystallizing and purifying by using a column to obtain the two-arm polyethylene glycol amine derivative intermediate containing Fmoc protected amino. Carrying out deprotection reaction on the obtained two-arm polyethylene glycol amine derivative intermediate containing Fmoc protected amino, removing Fmoc protecting groups, treating with 20% piperidine/DMF solution, removing solvent by rotary evaporation, dissolving with dichloromethane, precipitating with anhydrous ether, and recrystallizing with isopropanol to obtain branched polyethylene glycol amine derivative V-PEG with main chain containing a naked amino ₂(S20-3,9.85g, 75% yield):¹H NMR(CDCl₃)δ(ppm)：0.08(-Si(CH₃)₂-,12H),0.93(-SiC(CH₃)₃,18H),3.00-3.20(-C(O)NHCH₂CH₂O-,4H),3.42-3.76(-OCH₂CH₂O-,-OCH₂CH₂NH-,4H)。

step b: in a dry clean 1L round bottom flask, V-PEG is added₂Branched two-arm polyethylene glycol amine derivative S20-3(6.6KDa, 14.77g, 2.25mmol), benzenetricarboxylic acid compound S20-4(0.11g, 0.5mmol) and 400mL of tetrahydrofuran as a solvent were added DMAP (0.02g, 0.2mmol) and dicyclohexylcarbodiimide (DCC, 2.06g, 10.0mmol), and the reaction was stirred at room temperature for 12 hours. After the reaction was completed, insoluble matter was removed by filtration, concentrated and purified by silica gel column chromatography to obtain hexa-armed polyethylene glycol compound E20-1(7.54g, yield 76%) containing six TBS protected hydroxyl groups, and the hydrogen spectrum data of E20-1 was as follows:¹H NMR(CDCl₃)δ(ppm):0.08(-Si(CH₃)₂-,36H),0.93(-SiC(CH₃)₃,54H),3.42-3.76(-OCH₂CH₂O-,-OCH₂CH₂NH-,12H),4.10-4.25(-NHCH(CH₂O-)₂,12H)。

GPC measurement of the six-armed polyethylene glycol TBS protected hydroxy derivative E20-1 to determine M_n≈19.9kDa,PDI＝1.03。

Example 25: preparation of six-arm polyethylene glycol modified irinotecan derivative E21-1

The preparation process is as follows:

into a dry, clean 1L round-bottomed flask, hexa-armed polyethylene glycol H1-1(18.74g,2.0mmol), excess irinotecan propionic acid derivative S21-1(12.11g,18.0mmol) and the solvent dichloromethane (200mL) were added, DMAP (0.04g,0.3mmol) was added under ice-bath conditions, DCC (4.93g,24.0mmol) dissolved in 100mL dichloromethane was added dropwise to the reaction mixture, and after the addition was completed, the reaction was carried out at room temperature for 16H. After the reaction was completed, insoluble matter was removed by filtration, and the reaction mixture was concentrated and purified by column chromatography to obtain six-arm polyethylene glycol-modified irinotecan derivative E21-1(15.64g, yield 78%). The structure of E21-1 was determined by NMR.

GPC measurement of E21-1 to determine M_n≈10.0kDa,PDI＝1.02。

Example 26: biological testing of six-armed pegylated irinotecan

(1) Cytotoxicity assays

The cytotoxicity test of the six-arm pegylated irinotecan E21-1 is carried out by adopting an MTT staining method, a blank control combination positive control group is set in the experimental process, no medicament is added in the blank control group, only a culture medium is added in the blank control group, a single irinotecan medicament with a certain concentration is added in the positive control group, the six-arm pegylated irinotecan medicament with corresponding concentration is added in the experimental group, the medicament concentration is three gradient concentration points of 1nM, 10nM 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 10⁴Cells per well, 100. mu.L/well of cell suspension was seeded into 96-well plates. After inoculation, 4% CO at 37 ℃₂The 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 three 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 six-arm pegylated irinotecan with the corresponding concentration into the experimental group, wherein each concentration is 6 multiple wells. After 48h incubation of drug with cancer cells, 20. mu.L of MTT in PBS buffer at 5mg/mL was added to each well. After MTT and cancer cells are incubated for 4h, mixed liquor of a culture medium and MTT buffer solution is sucked away, DMSO (dimethylsulfoxide) is added into 150 mu L/hole, formazan serving as a purple crystal for dissolving living cells is dissolved, a 96-well plate is shaken up by gentle shaking in order to fully dissolve formazan, the shaking process cannot be performed too hard to avoid that a solution in a hole overflows to another hole to influence a test result, and after full shaking, an enzyme-labeling instrument is used for testing absorbance at 490 nm. The results of calculation drawing according to the measured absorbance values show that the irinotecan group and the six-arm pegylated irinotecan group have obvious cancer cell proliferation inhibiting effect on the six cancer cells compared with a blank control group; the six-arm pegylated irinotecan group also showed a stronger cancer cell proliferation-inhibiting effect on the above six cancer cells than the positive control group, i.e., the irinotecan alone group.

(2) Antitumor effect

Using animal transplantable tumor experimental method with H₂₂The liver cancer cells of the mouse are inoculated in the right axilla of the mouse to form solid tumors, and the solid tumors are respectively inoculated in the right axilla of the mouseAfter 2 days and 7 days, tail vein injection is carried out, and 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 both irinotecan and six-arm pegylated irinotecan have obvious tumor inhibition effects compared with the blank control, and the tumor inhibition rate of the six-arm pegylated irinotecan is significantly 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

1. A hexa-arm polyethylene glycol derivative is characterized in that the hexa-arm polyethylene glycol derivative has a structure shown in a general formula (1):

wherein A is₀Is a trivalent central structure;

PEG has the general formula

the hexa-arm polyethylene glycol derivative can exist stably or can be degraded; in the same molecule，A₀、A₁、L₂、L₀、G、(Z₂)_q-(Z₁)_q1Either, or the linking groups formed by either and adjacent groups, each independently, may be stable or degradable.

2. The hexa-armed polyethylene glycol derivative according to claim 1, wherein A is₀Comprises

Any one of the trivalent nuclear structures;

wherein R is₁Is a hydrogen atom, or is selected from any one of the following groups: 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 _1-20Alkyl, substituted aryl, substituted C_1-20Open-chain heterocarbyl, substituted heteroaralkyl; r₁Preferably 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, C group_1-10Halohydrocarbyl, haloacetyl or alkoxy substituted C_1-10An aliphatic hydrocarbon group; r₁Most preferably a hydrogen atom, a methyl group or an ethyl group;

wherein R is₃₇Is selected from C_1-20Hydrocarbyl, more preferably C_1-20Alkyl, most preferably methyl;

wherein M is₁₉Is an oxygen atom or a sulfur atom;

wherein M is₅、M₆、M₇、M₂₃Is a ring-forming atom, each of which is independently selected from any one of a carbon atom, a nitrogen atom, a phosphorus atom and a silicon atom; m₅、M₆、M₇、M₂₃The cyclic structure is 3-50 yuanA ring, preferably a 3-32 membered ring, more preferably a 3-18 membered ring, 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;

a is described₀Further preferably contains any of the following structures:

wherein Q is₅Is H atom, methyl, ethyl or propyl; when Q is₅When 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 integer j₂Selected from any one of 0, 1, 2, 3, 4, 5, 6;

a is described₀Still further preferred are compounds containing the above structure terminated with 3 identical divalent linking groups selected from the group consisting of oxy, thio, secondary amino, divalent tertiary amino and carbonyl; when participating in initiator molecules that constitute living anionic polymerization, are free of carbonyl groups, secondary amino groups;

a is described₀Still further preferred is any of the following structures:

3. the hexa-armed polyethylene glycol derivative according to claim 1, wherein A is₁Comprises

Any one of the trivalent nuclear structures; wherein A is₁Asterisks in the structure indicate that the asterisk ends point towards the trivalent center A₀Two non-star ends are the same and point to two L₂；

wherein M is₁₉Is an oxygen atom or a sulfur atom;

a is described₁Further preferably contains any of the following structures:

wherein Q is₅Is H atom, methyl, ethyl or propyl; when Q is₅When 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 A is₁Asterisks in the structure indicate that the asterisk ends point towards the trivalent center A₀With two identical non-star terminals pointing to two L₂；

A is described₁Further preferred are compounds containing the above structure terminated with 1, 2 or 3 identical or different divalent linking groups selected from oxy, thio, secondary amino, divalent tertiary amino and carbonyl groups; when participating in initiator molecules that constitute living anionic polymerization, are free of carbonyl groups, secondary amino groups;

a is described₁Further preferred is any of the following structures:

4. the hexa-armed 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.

5. The hexa-armed polyethylene glycol derivative according to claim 1, wherein n is₁、n₂、n₃、n₄、n₅、n₆The corresponding PEG branch chains are all polydisperse, and n₁≈n₂≈n₃≈n₄≈n₅≈n₆(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.

6. The hexa-arm polyethylene glycol derivative according to claim 1, wherein each of the six PEG chains has monodispersity, preferably n₁＝n₂＝n₃＝n₄＝n₅＝n₆(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.

7. The hexa-armed polyethylene glycol derivative according to claim 1,

the R is₀₁The interaction with the biologically relevant substance is selected from the group consisting of covalent bond formation, hydrogen bond formation, fluorescence, and targeting;

the R is₀₁Preferably 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 is₀₁Any 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 group, N-carbamoyl-3-imidazolyl group, N-carbamoyl-3-methylimidazolium iodide group, imido group, nitrone group, oxime group, ureido group, thioureido group, pseudoureido group; 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: cycloalkynylalkyl, cycloalkynheteroalkyl, conjugated dienyl, hybrid conjugated dienyl, 1,2,4, 5-tetrazinyl;

class Gb: azido, nitrile oxide groups, cyanooxide groups, cyano, isocyano, aldoximo, diazo, diazonium ions, azoxy, nitrilo imino, N-oxyaldoimino, 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.

8. The hexa-armed polyethylene glycol derivative according to claim 1,

the R is₀₁A 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:

or class B:

or class C:

or class D:

or class E:

or class F:

or class G:

class Ga:

or class Gb:

or class H:

or class I:

or class J:

wherein M is₅Is a ring-forming atom selected from any one of carbon atom, nitrogen atom, phosphorus atom and silicon atom; m₅The 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, Y₁Is 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, W₂Is F, Cl, Br or I;

wherein, W₃Is a leaving group selected from F, Cl, Br, I, PhS;

wherein the content of the first and second substances,

each is a cyclic structure containing a nitrogen atom, a nitrogen onium ion, a double bond, an azo, a triple bond, a disulfide bond, an anhydride, an imide, a diene on the ring backbone, 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 is₈Is a carbon atom, a nitrogen atom, a phosphorus atom or a silicon atom located on the ring; m ₈The number of ring-forming atoms of the ring is 4-50; preferably 4-32; more preferably 5 to 32;

wherein M is₂₂Is a carbon atom, a nitrogen atom, a phosphorus atom or a silicon atom on an alicyclic or alicyclic ring; m₂₂The number of ring atoms of the ring is 4, 5, 6, 7 or 8;

wherein R is₂Is a terminal group or a divalent linking group to which an oxygen or sulfur atom is bonded, selected from a hydrogen atom, R₂₁Or R₃Any one atom or group;

wherein R is₂₁Is a divalent linking group and participates in ring formation; r₂₁Is selected from C_1-20Alkylene, divalent C_1-20Heterohydrocarbyl, substituted C_1-20Alkylene, substituted divalent C_1-20Any divalent linking group or any two or any three of divalent linking groups in the heterohydrocarbon group; r₂₁Preferably methylene, ethylene, propylene, butylene, pentylene, hexylene, heptylene, octylene, nonylene, decylene, 1, 2-phenylene, benzylene, C_1-20Oxaalkylene, C_1-20Thiaalkylene group, C_1-20Any one group of azaalkylene and azaaralkyl, substituted form of any one group, any two or more of the same or different groups or substituted form thereofA combination of forms;

wherein R is₃Is a terminal group linked to an oxy or thio group, selected from C_1-20Hydrocarbyl radical, C_1-20Heterohydrocarbyl radical, C_1-20Substituted hydrocarbyl radical, C _1-20Any 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 is₄Is- (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 is₈、R₉、R₁₀、R₁₁、R₁₂Each independently is a hydrogen atom, a substituent atom or a substituent on a double bond (-C-), and R is in the same molecule₈、R₉、R₁₀、R₁₁、R₁₂May be the same as or different from each other; r₈、R₉、R₁₀、R₁₁、R₁₂Each independently selected from: hydrogen atom, fluorine atom, methyl group; in class E3, R₈Preferably methyl;

wherein R is₂₄Is a terminal group linked to a disulfide bond selected from: c_1-20Alkyl, aryl, hybrid phenyl;

wherein R is₂₇Is a substituent attached to azo selected from: phenyl, substituted phenyl or hybrid phenyl;

wherein R is₃₀Is a hydrocarbyl group selected from: c_1-20Alkyl, benzyl, phenyl ring hydrogen atoms by C _1-20A hydrocarbyl-substituted benzyl group;

wherein M is₁₉、M₂₀、M₂₁Each independently is an oxygen atom or a sulfur atom, and may be the same as or different from each other in the same molecule;

wherein, X₆Is a terminal group attached to the oxygen atom of the ester group and is selected from a hydroxyl protecting group or the group LG₄；LG₄Is selected from C_1-20Alkyl, aryl, aralkyl, C_1-20Heteroalkyl, heteroaryl, heteroaralkyl, C_1-20Alkylcarbonyl, arylcarbonyl, aralkylcarbonyl, C_1-20Heteroalkylcarbonyl, heteroarylcarbonyl, heteroaralkylcarbonyl, C_1-20Alkoxycarbonyl, aryloxycarbonyl, aralkyloxycarbonyl, C_1-20Alkylthio-carbonyl, arylthio-carbonyl, aralkylthiocarbonyl, C_1-20Alkylaminocarbonyl, arylaminocarbonyl, aralkylaminocarbonyl, C_1-20Heteroalkyloxycarbonyl, heteroaryloxycarbonyl, heteroaralkyloxycarbonyl, C_1-20Heteroalkylthio-carbonyl, heteroarylthio-carbonyl, heteroaralkylthio-carbonyl, C_1-20Heteroalkylaminocarbonyl, heteroarylaminocarbonyl, heteroarylalkylaminocarbonyl, C_1-20Alkylthio, arylthio, aralkylthiocarbonyl, C_1-20Heteroalkylthiocarbonyl, heteroarylthiocarbonyl, heteroarylalkylthiocarbonyl, C_1-20Alkoxythiocarbonyl, aryloxylthiocarbonyl, aralkyloxythiocarbonyl, C _1-20Alkylthio thiocarbonyl, arylthio thiocarbonyl, aralkylthio thiocarbonyl, C_1-20Alkylaminothiocarbonyl, arylaminothiocarbonyl, aralkylaminothiocarbonyl, C_1-20Heteroalkyloxythiocarbonyl, heteroaryloxythiocarbonyl, heteroarylalkoxythiocarbonyl, C_1-20Heteroalkylthio 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, X₁₁Is a terminal group attached to a carbonyl or thiocarbonyl group, selected from C_1-20An alkyl group;

wherein, X₁₂To a carbonate or thio groupTerminal carbonate group selected from C_1-20A hydrocarbyl group;

wherein, X₁₃Is a terminal group for attaching a sulfur group selected from: mercapto-protecting group, group LG₂；

Wherein LG is₂Selected 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 _1-10Any 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 is₃Is H atom or a group contributing to the inducing, conjugating effect of the electrons of the unsaturated bond, selected fromAny one atom or group 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;

Wherein Q is₅Is H atom, methyl, ethyl or propyl; when Q is₅When 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 is₆Is a hydrogen atom or a methyl group; q₇Is a hydrogen atom, a methyl group, a phenyl group or a substituted phenyl group; in the same molecule, Q₆And Q₇May be the same or different;

wherein Q is₈Is 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 is₈Is one or more; when more than 1, the structure is the same, or the combination of two or more different structures;

wherein Q is₁₁Is a substituent on the nitrogen atom of tetrazole, and is selected from any one of phenyl, substituted phenyl and aza-phenyl;

wherein PG₂Is a thiol protecting group, the protected thiol group being denoted as SPG₂Preferably any one of thioether, disulfide, silyl sulfide, thioester;

wherein PG₃Is an alkynyl protecting group, preferably a silicon group;

wherein PG₄Is a hydroxy protecting group, the protected hydroxy group being represented by OPG₄Any one of ether, silyl ether, ester, carbonate and sulfonate is preferable;

wherein PG ₅For protection of amino groupsThe protected amino group is represented by NPG₅Preferably any one of carbamate, amide, imide, N-alkylamine, N-arylamine, imine, enamine, imidazole, pyrrole and indole;

wherein PG₆Is a bishydroxy protecting group, and PG₆An acetal structure which forms a five-membered ring or a six-membered ring with two oxygen atoms; PG (Picture experts group)₆Is methylene or substituted methylene; wherein PG₆The 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 PG₈Protecting groups for orthocarbonic acid or orthosilicic acid.

9. The hexa-armed polyethylene glycol derivative according to claim 1,

said L₂、L₀(g＝1)、Z₁、Z₂Are all divalent linking groups, and are independent of each other, L in the same molecule₂、L₀(g＝1)、Z₁、Z₂May be the same as or different from each other;

L₂、L₀(g＝1)、Z₁、Z₂the structures of (a) are preferably each independently a linear structure, a branched structure or a cyclic-containing structure;

L₂、L₀(g＝1)、Z₁、Z₂each 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;

L₂、L₀(g＝1)、Z₁、Z₂Any one of, or any divalent linking group with an adjacent heteroatom group, is independently a linkage that can exist stablyA radical STAG or degradable linker 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-phenyl ring group, an aromatic heterocyclic group, a hetero-fused 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-phenyl ring group, a substituted aromatic heterocyclic group, a substituted hetero-fused 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, Sulfonamide, enamine, triazolyl, 4, 5-dihydroisoxazolyl, and the like,

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, Carbazolyl, thiocarbazoyl, azo, isoureido, isothioureido, allophanate, thioallophanate, guanidino, amidino, aminoguanidino, aminoamidino, imido, imidothioester, sulfonate, sulfinate, sulfonylhydrazino, sulfonylureido, maleimido, 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.

10. The hexa-arm polyethylene glycol derivative of claim 1, wherein g-1; the terminal branched groups G of the six-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.

11. The hexa-arm polyethylene glycol derivative of claim 1, 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 is₀-G comprises a structure selected from any one of the following groups: e₀,

Wherein, E is₀Contains any one of the following structures:

wherein Q is₅Is H atom, methyl, ethyl or propyl; when Q is₅When 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 E₀Preferably a capping structure containing a divalent linking group 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 E₀Any of the following structures is preferred:

Wherein, the asterisk marks in the structure, which indicates that the asterisk end points to the polyethylene glycol unit;

said E₀Preferably a trivalent backbone structure of an amino acid or a derivative thereof; wherein the amino acid is_LIs of type or_D-type; the amino acid or the derivative thereof is derived from any one of the following: serine, threonine, heminCystine, tyrosine, hydroxyproline, aspartic acid, glutamic acid, asparagine, glutamine, lysine, arginine, citrulline, histidine and tryptophan.

12. The hexa-armed polyethylene glycol derivative according to claim 1, wherein when k is 3, G is a tetravalent group; tetravalent G containing atoms CM₄And unsaturated bond CB₄Ring structure CC₄Any tetravalent core structure, or comprises two trivalent core structures; l is₀-G contains any of the following structures:

wherein, X₁Any one selected from methyl, ethyl, n-propyl, isopropyl, tert-butyl, pentyl, hexyl, allyl, trityl, phenyl, benzyl, nitrobenzyl, p-methoxybenzyl and trifluoromethylbenzyl; wherein the asterisks in the structure indicate that the asterisk ends point towards the polyethylene glycol unit.

13. The hexa-armed polyethylene glycol derivative according to claim 1, 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 valence of k + 1G contains 1 k +1 nuclear structure, or is formed by directly connecting and combining 2-k-1 3-k lower-valence groups or is formed by connecting and combining 1 or more than 1 divalent spacer groups L ₁₀Indirectly 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 nuclear structure, when k is more than or equal to 4 and contains a k + 1-valent nuclear structure, the k + 1-valent nuclear structure is a ring structure; when containing two or more L₁₀When L is₁₀May be the same as or different from each other; said directly or indirectlyG with the valence of k +1(k is more than or equal to 4) is formed by grafting and combining, and the combining mode is selected from any one of a comb combining mode, a tree combining mode, a branching combining mode, a hyperbranched combining mode and a ring combining 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:

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.

14. A process for the preparation of a hexa-armed polyethylene glycol derivative according to any one of claims 1 to 13, characterised in that it involves the steps of:

step one, adopting a hexahydroxy micromolecule containing six hydroxyl groups

Is stable under anionic polymerization conditions;

initiating ethylene oxide polymerization;

15. The process for preparing a hexa-armed polyethylene glycol derivative according to claim 14,characterized in that the hexahydroxy small molecule can be used as A through 1 residue₀The trifunctional small molecule and 3 residues can be A₁The trifunctional micromolecules are obtained through coupling reaction; wherein said residue may be A ₀The trifunctional small molecule of (a) contains three identical functional groups; the residue may be referred to as A₁The tri-functional small molecule of (a) contains two or three identical functional groups, when the residue can be A₁When the tri-functional small molecule contains only two identical functional groups, the different functional group ends and residues can be A₀Linking the trifunctional small molecules;

wherein, the residue may be A₀、A₁The trifunctional small molecules of (A) are preferably

Any one of, wherein, j₂Selected from any one of 0, 1, 2, 3, 4, 5, 6; r₁Is a hydrogen atom, or is selected from any one of the following groups: 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_1-20Alkyl, substituted aryl, substituted C_1-20Open-chain heterohydrocarbyl, substituted heteroaromatic hydrocarbyl.

16. The method for preparing the hexa-armed polyethylene glycol derivative according to claim 14, wherein the hexa-hydroxyl small molecule initiator IN- (OH)₆Selected from any one of the following structures:

wherein the integer j ₂Is any one of 0, 1, 2, 3, 4, 5 and 6.

17. A process for the preparation of a hexa-armed polyethylene glycol derivative according to any of claims 1 to 13, characterized in that it relates to 1 hexa-functionalized small molecule comprising six identical functional groups

Reacting with 6 linear double-end functionalized PEG derivative bilPEG molecules to obtain a six-arm polyethylene glycol derivative; wherein bilPEG is monodisperse or polydisperse; wherein functional groups at both ends of bilPEG may beThe same or different; wherein, F₁Contains a reactive functional group capable of reacting with a terminal functional group in the biliPEG to form a divalent linking group L₂；

18. The method for preparing the hexa-arm polyethylene glycol derivative of claim 17, wherein the hexa-functional small molecule can be A through 1 residue₀The trifunctional small molecule and 3 residues can be A ₁The trifunctional micromolecules are obtained through coupling reaction; wherein said residue may be A₀The trifunctional small molecule of (a) contains three identical functional groups; the residue may be referred to as A₁The tri-functional small molecule of (a) contains two or three identical functional groups, when the residue can be A₁When the tri-functional small molecule contains only two identical functional groups, the different functional group ends and residues can be A₀Linking the trifunctional small molecules;

wherein the residue may be A₀Preferably the trifunctional small molecule of (a) may be A in addition to the residue as described in claim 15₀The tri-functional small molecule can also be

The residue may be A₁The trifunctional small molecule of (a) may be A in addition to the residue as described in claim 15₁The tri-functional small molecule can also be

The hexafunctional small molecule is preferably any one of the following structures: IN- (OH)₆、

Wherein, the IN- (OH)₆Is any one of the hexahydroxy small molecules as defined in claim 16.

19. A process for the preparation of a hexa-armed polyethylene glycol derivative according to any of claims 1 to 13, characterized in that it involves 1 tri-functionalized small molecule containing three identical functional groups and 3 branched polyethylene glycol molecules V-PEG ₂The coupling reaction process of (1), reacting to obtain a hexa-arm polyethylene glycol derivative; wherein, the branched V-PEG₂Containing two identical branched PEG chain segments and one end of a main chain functional group derived from a branched core, the functional groups of the two PEG chain ends and the main chain end may be the same or different, and V-PEG₂The main chain functional group end of the molecule is connected with the trifunctional micromolecule; wherein, V-PEG₂Is monodisperse or polydisperse;

20. A bio-related substance modified by a six-arm polyethylene glycol derivative is characterized by having the following structural general formula:

wherein A is₀Is a trivalent central structure;

A₁is a trivalent branched structure, three A₁Are all the same; a. the₁Connection A₀And two L are led out₂And A is₁Middle two L₂The two ends of (a) are the same; a. the₀And A₁Covalently attached moieties, excluding A₀And A₁The respective part of the branched core is denoted L _A0A1，L_A0A1Is a divalent linking group containing an ether bond, a thioether bond, an ester bond, an amide bond, a carbonate bond, a urethane bond or a urea bond;

PEG has the general formula

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 is₀Is a divalent linking group which connects the PEG chain segment with the terminal branching group G;

when g is 0, k is 1, L₀G is not present;

when G is 1, G is present, L₀May be present or absent, k is 2-250;

EF may be expressed as ED (structure is

) Or EF₁(structure is

) And D is not equal to E₀₁(ii) a Wherein q and q are₁Each independently is 0 or 1; z₁、Z₂Each independently is a divalent linking group; wherein D is a residue formed by the reaction of the modified bio-related substance and the hexa-arm polyethylene glycol derivative; e₀₁Is selected from R₀₁Protected R ₀₁Deprotected R₀₁Or blocked R₀₁；R₀₁Is a reactive group capable of reacting with a biologically relevant substance; l is a linking group formed after a reactive group in the six-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_DK is not more than k, k of each branched chain in the same molecule_DEach independently the same or different, and the sum of the numbers of D in any one of the hexa-armed polyethylene glycol derivative molecules (N)_D) At least 1, preferably at least 6; when G is 1, G- (EF)_kCan be expressed as

the bio-related substance modified by the hexa-arm polyethylene glycol derivative can exist stably or can be degraded; in the same molecule, A₀、A₁、L₂、L₀、G、(Z₂)_q-(Z₁)_q1、(Z₂)_qAny of L, or any of the linkages with adjacent groups, is independently stabilizableEither absent or degradable.

21. The hexa-armed polyethylene glycol derivative-modified bio-related substance according to claim 20, wherein when g is 0, the structure is represented by formula (3):

or when the g is 1 and the content of D is 100%, the structure is shown as the formula (4):

22. The hexa-armed polyethylene glycol derivative modified bio-related substance according to claim 20, wherein k at the end of six PEG chains in the same molecule_DAll satisfy k being more than or equal to 1_DK, i.e. at least one D is attached to each branch chain;

preferably, k is at the end of six PEG chains in the same molecule_DAll satisfy k, i.e., all of the terminal reactive sites in the six-armed polyethylene glycol derivative molecule are each independently linked to a D.

23. The hexa-armed polyethylene glycol derivative modified bio-related substance of claim 20, 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%.

24. The bio-related substance modified by the hexa-armed polyethylene glycol derivative according to any one of claims 20 to 23, wherein L corresponding to six PEG chain ends in the same molecule is not identical, or L corresponding to six PEG chain ends is identical; 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 eight PEG chains of the same molecule have the same stability, i.e. are both stably present or both 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, azacyclic thioacetal, thiocyclic ketal, azacyclic ketal, thiocyclic ketal, imine bond, hydrazone bond, acylhydrazone bond, oxime bond, sulfoximine group, semicarbazone bond, thiosemicarbazone bond, hydrazon group, hydrazide group, thiocarbonyl group, azocarbohydrazide group, thioazocarbohydrazide group, hydrazinoformate group, hydrazinothiocarbamate group, carbazide group, thiocarbcarbazide group, azo group, isourea group, isothiourea group, thiosemicarbazide group, thio, 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.

25. The hexa-armed polyethylene glycol derivative modified bio-related substance according to claim 20, wherein the hexa-armed polyethylene glycol derivative modified bio-related substance meets any one of the following conditions:

(1) g-0, having stable hexavalent centers

stabilized-O- (Z)₂)_q-L-；

(2) g-0, having stable hexavalent centers

degradable-O- (Z)₂)_q-L-；

(3) g-0, having degradable hexavalent centers

degradable-O- (Z)₂)_q-L-；

(4) g 1, having stable hexavalent centers

(5) g 1, having stable hexavalent centers

(6) g 1, having stable hexavalent centers

(7) g 1, having degradable hexavalent centers

(8) g 1, having degradable hexavalent centers

(9) g 1, having degradable hexavalent centers

(10) g 1, having stable hexavalent centers

(11) g 1, having degradable hexavalent centers

(12) g-0, having stable hexavalent centers

stabilized-O- (Z)₂)_q-, degradable L;

(13) g 1, having stable hexavalent centers

stabilized-O-L₀-G-[(Z₂)_q-]_kA degradable L;

(14) g-0, having stable hexavalent centers

stabilized-O- (Z)₂)_qDegradable L-D;

(15) g 1, having stable hexavalent centers

stabilized-O-L₀-G-[(Z₂)_q-]_kDegradable L-D.

26. The hexa-armed polyethylene glycol derivative modified biologically-relevant substance according to claim 20, wherein said biologically-relevant substance is selected from any one of: 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; allowing a target molecule, an adjunct or a delivery vehicle to bind to the biologically-relevant substance before or after binding to the six-armed 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.

27. The hexa-armed polyethylene glycol derivative modified bio-related substance according to claim 20, wherein the bio-related substance is preferably a small molecule drug selected from the group consisting of: biologically relevant substances with molecular weight not more than 1000Da and small molecular mimicry or active fragment of any biologically relevant substance; 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.

28. The hexa-armed polyethylene glycol derivative modified bio-related substance according to claim 20, 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, phenylpropanoids, anthracyclines, and aminoglycosides;

The small molecule drug is preferably in any one of the following therapeutic 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 the hydrochloride salt; the derivatives comprise molecular modified derivatives, glycosides, nucleosides, amino acids and polypeptide derivatives.