CN113979895B - Self-degradable polymer with controllable precise sequence and preparation method and application thereof - Google Patents

Self-degradable polymer with controllable precise sequence and preparation method and application thereof Download PDF

Info

Publication number
CN113979895B
CN113979895B CN202010651204.7A CN202010651204A CN113979895B CN 113979895 B CN113979895 B CN 113979895B CN 202010651204 A CN202010651204 A CN 202010651204A CN 113979895 B CN113979895 B CN 113979895B
Authority
CN
China
Prior art keywords
formula
self
polymer
under
degradation
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN202010651204.7A
Other languages
Chinese (zh)
Other versions
CN113979895A (en
Inventor
刘世勇
石强强
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
University of Science and Technology of China USTC
Original Assignee
University of Science and Technology of China USTC
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by University of Science and Technology of China USTC filed Critical University of Science and Technology of China USTC
Priority to CN202010651204.7A priority Critical patent/CN113979895B/en
Publication of CN113979895A publication Critical patent/CN113979895A/en
Application granted granted Critical
Publication of CN113979895B publication Critical patent/CN113979895B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C271/00—Derivatives of carbamic acids, i.e. compounds containing any of the groups, the nitrogen atom not being part of nitro or nitroso groups
    • C07C271/06—Esters of carbamic acids
    • C07C271/08—Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms
    • C07C271/26—Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms with the nitrogen atom of at least one of the carbamate groups bound to a carbon atom of a six-membered aromatic ring
    • C07C271/28—Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms with the nitrogen atom of at least one of the carbamate groups bound to a carbon atom of a six-membered aromatic ring to a carbon atom of a non-condensed six-membered aromatic ring
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
    • A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C269/00—Preparation of derivatives of carbamic acid, i.e. compounds containing any of the groups, the nitrogen atom not being part of nitro or nitroso groups
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C269/00—Preparation of derivatives of carbamic acid, i.e. compounds containing any of the groups, the nitrogen atom not being part of nitro or nitroso groups
    • C07C269/02—Preparation of derivatives of carbamic acid, i.e. compounds containing any of the groups, the nitrogen atom not being part of nitro or nitroso groups from isocyanates with formation of carbamate groups
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/62—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating the ionisation of gases, e.g. aerosols; by investigating electric discharges, e.g. emission of cathode
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/50—Improvements relating to the production of bulk chemicals
    • Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Animal Behavior & Ethology (AREA)
  • Veterinary Medicine (AREA)
  • General Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Engineering & Computer Science (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Public Health (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Electrochemistry (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Polyethers (AREA)
  • Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)

Abstract

The invention provides a self-degradable polymer with controllable precise sequence, which has a structure shown in a formula I. The self-degradable polymer with controllable precise sequence consists of three parts: a) the Trigger depolymerization end is Trigger end, b) degradable self-degradation segment, c) dendritic Tag end composed of glycol chain segment or carbon chain. The number of repeating units of the polymer is determined, the sequence is controllable, and the precise polymer can be spontaneously depolymerized by electronic rearrangement after the trigger element is removed under the corresponding stimulation condition. The invention develops a new method for determining an accurate polymer sequence by using a macromolecular mass spectrum by utilizing the characteristic of the accurate self-degradation polymer in catalytic degradation. On the other hand, the invention also determines the sequence of the accurate self-degradation polymer through a macromolecular tandem mass spectrum, and screens out a structure capable of directly reading according to a secondary mass spectrum. The polymer can also be applied to the application fields of novel information storage carriers, anti-counterfeiting, information encryption and the like.

Description

Self-degradable polymer with controllable precise sequence and preparation method and application thereof
Technical Field
The invention relates to the technical field of precise macromolecules, in particular to a self-degradable polymer with controllable precise sequence and a preparation method and application thereof.
Background
Biological macromolecules such as nucleic acids and proteins, because of the defined monomer position and structure on their polymer chains, play a decisive role in the storage of their genetic information and in specific biological functions. The exploration of the link between polymer structure and function, mimicking these natural macromolecules of defined structure, has generated great interest to researchers. The precise synthesis of macromolecules, namely the realization of precise regulation and control of a macromolecule chain structure, is very important in the aspects of researching the relationship between a macromolecule structure and performance and developing new materials, and is also a great challenge. Unlike the synthesis of small molecules, the precise synthesis of macromolecules mainly faces the following four problems: 1) How accurately the molecular weight is controlled; 2) How to control the order of arrangement of the monomer units; 3) How to realize efficient and highly selective post-modification; 4) How to purify a polymer efficiently.
The existing precise polymers can be subjected to successive tandem depolymerization after triggering and dissociating the protective elements at specific positions, so as to release small molecule construction elements. The concept of triggering degradable polymers was first proposed by Shabat, followed by the development of dendritic and hyperbranched self-degrading polymers.
The precise self-degradable polymer has strong attraction because the structure is controllable, the degradation time can be controlled, and the time and the space are perfectly combined together for the property of specific environmental response. But currently there is less research due to the greater challenges in their synthesis.
Disclosure of Invention
In view of this, the technical problem to be solved by the present invention is to provide a self-degradable polymer with controllable precise sequence, a preparation method and an application thereof, and more specifically, to provide a linear polymer with controllable precise sequence and a synthesis thereof, wherein the linear polymer is based on carbamate linkage and causes main chain degradation through electronic rearrangement after triggering; according to two different principles, the molecule sequence is determined by utilizing a macromolecular mass spectrum technology, and the direct reading of the stored information in the polymer is realized by optimizing the structure of the polymer.
The invention provides a self-degradable polymer with controllable precise sequence, which has a structure shown in a formula I:
Figure BDA0002575044570000021
n is any integer of 1 to 12; in some embodiments, n is 1, 2, 3,4, 5, 6, 7, 8, 9, 10, 11, or 12;
Figure BDA0002575044570000022
to be triggeredA moiety that is responsive to ultraviolet light, visible light, an acidic environment, an oxidizing or reducing environment;
Figure BDA0002575044570000023
the molecular label is a dendritic structure, and a branching unit of the dendritic structure is a hydrophilic polyethylene glycol structure or a hydrophobic alkyl chain; />
Figure BDA0002575044570000024
Is a structural unit and is a self-degradation part of the self-degradation polymer with controllable precise sequence;
wherein R and R 1 Independently selected from H, F or Cl; further preferably, R = R 1 ＝H；
Or R is H, R 1 Selected from NO 2 Alkyl with 1 to 10 carbon atoms, alkoxy with 1 to 10 carbon atoms, F, cl, br or I;
further preferably, R is H, R 1 Selected from NO 2 C1-C5 alkyl, C1-C5 alkoxy, F, cl, br or I;
more preferably, R is H, R 1 Selected from NO 2 Methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, tert-butoxy, F, cl, br or I;
when n >1, the structures of the respective structural units may be the same or different.
For example, R = R among all the structural units in the precise sequence-controllable self-degrading polymer 1 ＝H。
Or in one or more structural units in the self-degradable polymer with controllable precise sequence, R = R 1 = H; in one or more other structural units, R is H, R 1 Is a methyl group.
The different structural units can be combined randomly to form the precise self-degradation polymer with different sequences.
In the present invention, the number of carbon atoms in the polyethylene glycol structure is preferably 2 to 32.
The number of carbon atoms of the polyethylene glycol structure refers to the number of carbon atoms contained in one polyethylene glycol chain of the dendritic polyethylene glycol structure.
In the present invention, the number of carbon atoms in the alkyl chain is preferably 1 to 20.
The number of carbon atoms of the alkyl chain refers to the number of carbon atoms of a carbon chain in the dendritic alkyl chain.
In the invention, after the trigger element is removed, the self-degradable polymer with controllable precise sequence undergoes spontaneous sequential depolymerization to release carbon dioxide and small molecular structural units.
According to the invention, preferably, the
Figure BDA0002575044570000031
Selected from any of the following structures:
Figure BDA0002575044570000032
/>
wherein R is 2 H or alkoxy with 1 to 20 carbon atoms; more preferably, R 2 Is H or alkoxy having 10 to 20 carbon atoms, and in some embodiments of the invention, R2 is (7R, 11R) -3,7,11, 15-tetramethylhexadecyloxy.
R 3 Is H or CH 3 ；
X is O or S.
In the structure, T1 and T2 have responsiveness to ultraviolet light; t3 is responsive to reducing substances such as triphenylphosphine; t4, T5, T6 are responsive to visible light; t7 is responsive to Glutathione (GSH), dithiothreose (DTT), and the like; t8 is acid responsive; t9 is responsive to active oxygen.
The above-mentioned
Figure BDA0002575044570000033
Selected from any of the following structures:
Figure BDA0002575044570000041
wherein R' is selected from any one of the following structures a to j:
Figure BDA0002575044570000042
m is an integer of 1 to 16, more preferably an integer of 1 to 10, and still more preferably an integer of 3 to 8; in some embodiments of the invention, m is 3,4, 5, 6, 7, 8;
n 1 is an integer of 1 to 19, more preferably an integer of 1 to 10, still more preferably 1, 2, 3,4, 5, 6, 7, 8, 9, 10;
y is CH 3 Trityl (Trt) or benzyl (Bn).
The invention provides a method for defining the digital macromolecules of the self-degradation polymer with controllable precise sequences, wherein in the self-degradation segment, R = R 1 H is defined as 0 in binary; r = H, R 1 ＝CH 3 Defined as a1 in binary.
The invention provides two preparation methods of the self-degradation polymer with controllable precise sequences, which comprise the following steps:
one method is modular synthesis from a Trigger end by using an iterative growth method to a Tag end, and specifically comprises the steps of converting benzoyl azide of a compound with a structure shown in a formula (1) into phenyl isocyanate under a heating condition, reacting the phenyl isocyanate with hydroxyl of a Trigger element in a solvent under the action of a catalyst DBTL, removing tert-butyl dimethylsilane for protecting the hydroxyl, continuously repeating the two reactions until a target polymerization degree is reached, and finally blocking a Tag molecule.
The other method is modular synthesis from Tag end to Trigger end by utilizing orthogonal chemical reaction, specifically, acyl azide (amide) on a Tag molecule reacts with a compound with a structure of formula (2) and benzamide at the left end in a solvent under the action of a catalyst, the reaction is continuously repeated, and finally a Trigger element is blocked.
The mechanism of the above preparation method is based on the following orthogonal chemical reactions:
the phenyl acyl azide can be converted into isocyanate under the heating condition, and can be efficiently coupled with hydroxyl under the catalysis of DBTL; benzamide can be converted into isocyanate in the presence of an iodine reagent, and can be efficiently coupled with hydroxyl under the catalysis of DBTL; the two conversions do not interfere with each other and the reactions are orthogonal. Wherein the iodine reagent may be of the structure:
Figure BDA0002575044570000051
in a preferred embodiment the iodine reagent is PhINTs.
Specifically, the invention provides a preparation method of the self-degradation polymer with controllable precise sequence, which comprises the following steps:
s1) heating and converting an intermediate shown in a formula (1) to convert an acyl azide group into an isocyanate group to obtain a phenyl isocyanate compound;
under the action of a catalyst, reacting a phenyl isocyanate compound with the hydroxyl of a triggering element hydroxyl compound to obtain a structure shown in a formula (2):
s2) removing a protecting group of hydroxyl, namely tert-butyl dimethyl silane group to obtain a compound shown as a formula (3);
s3) heating and converting the intermediate shown in the formula (1) to convert acyl azide groups into isocyanate groups to obtain phenyl isocyanate compounds;
under the action of a catalyst, reacting a phenyl isocyanate compound with a structure shown in a formula (3);
s4) repeating the steps S2-S3) for 0-11 times;
s5) removing a protecting group of hydroxyl, namely tert-butyl dimethyl silane, reacting with a compound shown as a formula (4), and blocking a label molecule to obtain a self-degradable polymer with a controllable precise sequence;
Figure BDA0002575044570000061
the T, tag, R and R 1 The same scope is defined above, and is not described herein.
The catalyst is preferably PhINTs.
The invention provides another preparation method of a self-degradation polymer with controllable precise sequence, which comprises the following steps:
a) Reacting acyl azide groups of the label molecules shown in the formula (4) with hydroxyl groups of the compound shown in the formula (5) under a heating condition to obtain a compound shown in the formula (6);
b) Converting an amide group of the compound shown in the formula (6) into an isocyanate group under the action of an iodine reagent, and then reacting the isocyanate group with a hydroxyl group of the compound shown in the formula (7) under the action of a normal-temperature catalyst to obtain a compound shown in the formula (8);
c) Reacting acyl azide groups of the compound shown in the formula (8) with hydroxyl groups of the compound shown in the formula (5) under a heating condition, converting amide groups of obtained products into isocyanate groups under the action of an iodine reagent, and then reacting with the hydroxyl groups of the compound shown in the formula (7) under the action of a normal-temperature catalyst;
d) Repeating the step C) for 0 to 11 times;
e) When the end of the obtained intermediate is an amide group, the intermediate is converted into an isocyanate group under the action of an iodine reagent;
when the end of the obtained intermediate is acyl azide group, the acyl azide group is converted into isocyanate group under the heating condition;
then under the action of a catalyst, reacting with a trigger element shown in a structure of a formula (9) to terminate the end, so as to obtain a self-degradable polymer with controllable precise sequence;
Figure BDA0002575044570000071
t, tag, R 1 The same scope is defined above, and is not described herein.
The catalyst is preferably PhINTs.
Triggering type self-degradation of the polymer can be initiated after the Trigger is removed under the action of an external field, intermediates generated by degradation of the self-degradation polymer with controllable precise sequences are detected by MALDI-TOF at different time points to obtain molecular weight information of all the generated intermediates, and sequence information is obtained by analyzing the difference between the molecular weights. Meanwhile, the accurate self-degradation polymer can be directly subjected to macromolecular tandem mass spectrometry without triggering to obtain the sequence information of the accurate self-degradation polymer.
The invention provides the application of the self-degradable polymer with controllable precise sequence or the self-degradable polymer with controllable precise sequence prepared by the preparation method in the time-space controllable release of drugs and the time-space controllable conduction of signals.
The invention provides the application of the self-degradable polymer with controllable precise sequence or the self-degradable polymer with controllable precise sequence prepared by the preparation method in a novel polymer molecule sequencing technology.
The invention provides the self-degradation polymer with controllable precise sequence or the self-degradation polymer with controllable precise sequence prepared by the preparation method, which can selectively break bonds in a tandem mass spectrum and can be applied to novel information storage carriers, product anti-counterfeiting and information encryption.
The present invention also provides a method for tracking the degradation of self-degrading polymers using Gel Permeation Chromatography (GPC). Selecting a proper solvent system according to the structure of the obtained self-degradable polymer, simultaneously adding a trapping agent of an active intermediate generated in the degradation process, triggering and removing a Trigger group in a corresponding external field, carrying out decarboxylation reaction on the polymer, carrying out electronic rearrangement on a main chain to release a structural unit, generating spontaneous degradation, taking equal volume of samples in the system by using a microsyringe at different time points, and adding excessive trifluoroacetic acid to terminate the reaction; the sample is pumped to be dry and added into the same amount of mobile phase for solution sampling, and then the degradation condition of the self-degradation polymer can be tracked accurately.
In a preferred embodiment, the polymer concentration is 3mg/mL to 10mg/mL and the sample volume is 50. Mu.L.
The invention provides a sample preparation method for characterizing an accurate self-degradation polymer by using MALDI-TOF, which comprises the following steps: dissolving trans-2- [3- (4-tert-butyl benzene) -2-methyl-2-pentene ] malononitrile (DCTB) serving as a matrix in chromatographic pure dichloromethane, wherein the concentration is 20mg/mL; sodium trifluoroacetate is selected as an auxiliary salt to be dissolved in chromatographically pure methanol, and the concentration is 10mg/mL; mixing the matrix and the auxiliary salt solution 10 in a volume ratio of 1 to obtain a matrix solution; dissolving a sample to be detected by using chromatographically pure tetrahydrofuran, wherein the concentration is 5mg/mL; during sample preparation, a sandwich sample preparation method is adopted, namely, 2 mu L of matrix solution is paved on a target plate firstly until the matrix solution is completely volatilized, 2 mu L of solution to be detected is paved on the matrix until the solvent is completely volatilized, and finally 2 mu L of matrix solution is paved on the surface of a sample again and dried overnight.
The invention provides a method for sequencing by utilizing a macromolecule mass spectrum by utilizing the triggering degradation characteristic of an accurate self-degradation polymer. Selecting a proper solvent system according to the structure of the obtained accurate self-degradable polymer, triggering and removing a Trigger group in a corresponding external field, then carrying out decarboxylation reaction on the polymer, carrying out electronic rearrangement on a main chain to release a structural unit, carrying out spontaneous degradation to generate a degradation intermediate, sampling at a specific time point, terminating degradation by trifluoroacetic acid, preparing a sample according to the method, testing a macromolecular mass spectrum, obtaining the molecular weight of the intermediate with different degradation times, and comparing the molecular weight difference to obtain the sequence structure of the accurate self-degradable polymer.
In the invention, the self-degradation polymer with controllable precise sequence can realize direct reading of molecular information, and the Tag end of the self-degradation polymer is a dendritic glycerol chain which has strong binding capacity to auxiliary ions such as sodium ions or potassium ions.
The invention also provides a direct sequence determination method, which is used for accelerating the selected molecular ions in a macromolecular mass spectrum LIFT mode and selectively breaking the molecular ions at the specific position of the molecular skeleton to obtain the fragment peak of the molecules.
Compared with the prior art, the invention provides a self-degradable polymer with controllable precise sequence, which has a structure shown in a formula I. The self-degradable polymer with controllable precise sequence consists of three parts: a) the Trigger depolymerization end is Trigger end, b) degradable self-degradation segment, c) dendritic Tag end composed of glycol chain segment or carbon chain.
The number of repeating units of the polymer is determined, the sequence is controllable, and the precise polymer can be spontaneously depolymerized by electronic rearrangement after the trigger element is removed under the corresponding stimulation condition. The invention develops a new method for determining an accurate polymer sequence by using a macromolecular mass spectrum by utilizing the characteristic of the accurate self-degradation polymer in catalytic degradation. On the other hand, the invention also determines the sequence of the accurate self-degradation polymer through a macromolecule tandem mass spectrum, and screens out a structure which can be directly read according to a secondary mass spectrum. The self-degradable polymer with controllable precise sequence can also be applied to the application fields of novel information storage carriers, anti-counterfeiting, information encryption and the like.
Drawings
FIG. 1 is an HPLC chromatogram for the validation of the orthogonal reaction in example 1 of the present invention;
FIG. 2 is a nuclear magnetic hydrogen spectrum diagram of monomers and dimers constructed when the precise self-degradable polymer is synthesized from a trigger element end to a label molecule end;
FIG. 3 is a nuclear magnetic hydrogen spectrum of a monomer and a dimer constructed when a precise self-degradable polymer is synthesized from a label molecule end to a trigger element end according to the present invention;
FIG. 4 is a nuclear magnetic hydrogen spectrum of a tag molecule designed according to the present invention;
FIG. 5 is a nuclear magnetic resonance hydrogen chromatogram, a gel permeation chromatogram and a macromolecular mass chromatogram of the precise sequence controllable self-degradable polymer prepared in example 5 of the present invention;
FIG. 6 is a NMR chart, a gel permeation chromatograph and a macromolecular mass spectrum of the precise sequence controllable self-degradable polymer prepared in example 6 of the present invention;
FIG. 7 is a gel permeation chromatogram of the tracking degradation of a precise sequence controllable self-degradable polymer prepared by the present invention;
FIG. 8 is a diagram of the macromolecular mass spectrum and its corresponding mass spectrum peak attributed to the tracking degradation of the precise sequence controllable self-degradable polymer prepared by the present invention in FIG. 7;
FIG. 9 is a macromolecular primary mass spectrum and a secondary mass spectrum of the precise sequence controllable self-degradable polymer prepared by the invention;
FIG. 10 shows the macromolecular primary mass spectrum and secondary mass spectrum of the precise sequence controllable self-degradable polymer prepared by the present invention.
Detailed Description
In order to further illustrate the present invention, the following examples are provided to describe the precise sequence controllable self-degradable polymers provided by the present invention and their preparation and application in detail.
Example 1: validation of orthogonality of the reaction of benzoyl azide with the conversion of benzamide to isocyanate
The following two substances are selected as verified model compounds, one is tert-butyldimethylsilane protected p-hydroxymethylbenzamide, and the other is tert-butyldimethylsilane protected p-hydroxymethylbenzoyl azide; mixing the two solutions under the condition of equal amount of substances, reacting with excessive benzyl alcohol under the condition of heating and the existence of iodine reagent, and verifying the reaction condition of the materials by using a High Performance Liquid Chromatography (HPLC) method.
The specific reaction is as follows:
Figure BDA0002575044570000101
mixing OTBS-0-CON 3 (50mg,0.17mmol,1equiv.)，OTBS-0-CONH 2 (45mg, 0.17mmol, 1equiv.), benzyl alcohol (36.8mg, 0.34mmol, 2equiv.), DBTL (dibutyltin dilaurate) in catalyst equivalent weight are respectively added into a dry round-bottom flask, 10mL of anhydrous toluene is added for azeotropic water removal, the process is repeated for three times, nitrogen protection is carried out after the water removal is finished, and 2mL of dry toluene is added as a reaction solvent to heat the system to 85 ℃ for reaction for 4 hours.
Figure BDA0002575044570000102
Mixing OTBS-0-CON 3 (50mg,0.17mmol,1equiv.)，OTBS-0-CONH 2 (45mg, 0.17mmol, 1equiv.), phINTs (76.1mg, 0.204mmol, 1.2equiv.) were added into the round-bottom flask 1, 10mL of anhydrous toluene was added for azeotropic removal of water, the process was repeated three times, and 1mL of anhydrous toluene was added under nitrogen protection; benzyl alcohol (36.8mg, 0.34) was weighed outmmol,2 equiv.), adding 10mL of toluene into a flask 2 for azeotropic dehydration, repeating for three times, protecting with nitrogen, adding 1mL of anhydrous toluene, transferring the solution in the flask 2 into the flask 1 by using a double-needle transfer technology, and reacting for 4 hours at normal temperature.
After the two reactions, the reaction conditions were checked at a detection wavelength of 225nm under conditions of gradient elution from acetonitrile/water of 7/3 (volume ratio) to acetonitrile/water of 9/1. The results are shown in FIG. 1.
It can be seen from FIG. 1 that benzoyl azide is converted to phenyl isocyanate under heating, reacting with hydroxyl, while benzamide does not participate in the reaction; under the action of PhINTs and under the reaction condition of normal temperature, benzamide is converted into phenyl isocyanate to react with hydroxyl, benzoyl azide does not participate in the reaction, and the two reaction conditions are combined to show that the conditions for converting the benzoyl azide and the benzamide into the phenyl isocyanate are orthogonal.
Example 2: the first step of synthesizing a self-degradable polymer building module from a trigger element end to a molecular tag end is to prepare a monomer building element, taking the synthesis of the monomer building element with the following side group as methyl as an example:
Figure BDA0002575044570000111
one end of monomer building element molecule benzyl hydroxyl group is protected by tert-butyl dimethyl silane, and the other end is benzoyl azide.
The preparation method comprises the following steps: 1) 3-methyl-4-bromobenzoic acid (10g, 46.5mmol, 1equiv.) is weighed into a dry round-bottom flask, a borane tetrahydrofuran solution (93mL, 93mmol, 2equiv.) with the concentration of 1M is slowly added into the flask by a glass syringe under the conditions of ice bath and nitrogen protection, and after the dropwise addition is completed, the flask is heated by nitrogen protection and oil bath for 55 ℃ reaction overnight. After the reaction is finished, methanol is added to quench the reaction, the reaction solvent is dried in a spinning mode, then ethyl acetate is added to dissolve the reaction solvent, deionized water is used for washing, the organic phase is dried by anhydrous magnesium sulfate, 8.8g of 4-bromo-3-methylbenzyl alcohol with reduced carboxyl is obtained through drying in a spinning mode, and the yield is as follows: 95 percent.
2) Then 4-bromo-3-methylbenzyl alcohol (8g, 39.8mmol, 1equiv.), imidazole (3.3g, 47.8mmol, 1.2equiv.) were placed in a dry round-bottom flask, and 30mL of dichloromethane was added thereto and dissolved; weighing tert-butyldimethylsilyl chloride (9g, 59.7mmol, 1.5equiv.) into a 100mL constant pressure dropping funnel, adding 30mL of dichloromethane into the constant pressure dropping funnel, dissolving the dichloromethane, slowly dropping the raw materials in the constant pressure dropping funnel into a round bottom flask under the ice bath condition, and reacting overnight; the organic phase is dichloromethane, the reaction system is washed for three times by deionized water and is dried by spinning to obtain 11.3g of product, and the yield is 90%.
3) Adding the product (2g, 6.3mmol, 1equiv.) obtained in the last step into a round-bottom flask, then carrying out azeotropic dehydration on toluene for three times, adding 10mL of anhydrous tetrahydrofuran under the protection of nitrogen, cooling for 10min at-78 ℃, then slowly adding 12.7mL of n-butyllithium solution (1M), stirring the system for 30min, then adding anhydrous DMF (2.4 mL,31.5mmol, 5equiv.), reacting for 30min at-78 ℃, and then transferring to room temperature to continue reacting for 2h; adding saturated ammonium chloride solution to quench the reaction after the reaction is finished; removing tetrahydrofuran under reduced pressure, extracting with ethyl acetate, washing with saturated ammonium chloride for three times, spin-drying solvent, and performing column chromatography with petroleum ether-ethyl acetate as eluent to obtain pure product 1.4g with yield of 85%.
4) The product (1.4 g,5.3mmol, 1equiv.) obtained in 3) above, 2-methyl-2-butene (0.97g, 31.8mmol,6 equiv.) were placed in a round-bottomed flask, then 10mL of acetone was added to dissolve it, 6 equivalents of sodium chlorite, 1.4g of sodium dihydrogen phosphate dihydrate and 2.9g were weighed in an erlenmeyer flask, 10mL of deionized water was added to dissolve them, then the flask was slowly dropped into the round-bottomed flask, the reaction was stopped after vigorous stirring for 30min, ethyl acetate was added to spin-dried acetone, the organic phase was washed three times with a saturated ammonium chloride solution, then washed 3 times with a saturated sodium thiosulfate solution, finally washed three times with a saturated common salt solution, dried over anhydrous magnesium sulfate and then spin-dried to obtain 1.36g of the product, yield: 92 percent.
5) The product (1g, 3.6mmol, 1equiv.) obtained in the above 4) was dissolved in a dry round-bottom flask by adding 2mL of tetrahydrofuran, and one equivalent of 361mg of triethylamine was added; DPPA (1.1g, 3.6mmol, 1.1equiv.) was weighed and slowly added dropwise into a round-bottomed flask under ice bath conditions, and then the mixture was invertedThe reaction time is 4 hours; after the reaction is finished, the solvent is dried by decompression and spin, the product 880mg is obtained by taking petroleum ether-ethyl acetate as eluent for column chromatography, the yield is 80 percent, and the monomer is marked as OTBS-1-CON 3 The structure is characterized by nuclear magnetic hydrogen spectrum, and the result is shown in figure 2, a).
On the other hand, similar monomer building units without substituent groups on the benzene ring are marked as OTBS-0-CON 3 Synthesized according to document US20170252458 A1.
OTBS-0-CON 3 The structure is as follows:
Figure BDA0002575044570000121
the second step is to prepare the building unit of the dimer, and the specific synthetic route is as follows:
Figure BDA0002575044570000122
r or R 1 Can arbitrarily represent H or CH 3 (ii) a The benzoyl azide is completely converted into isocyanate under the heating condition, and then the temperature of the system is reduced to room temperature or lower to react with hydroxyl under the catalysis of DBTL.
The preparation method comprises the following steps: 1) Placing the methyl-containing monomer building element (2g, 6.5mmol, 1equiv.) prepared in the first step into a round-bottom flask, dissolving the methyl-containing monomer building element in 4mL of methanol, then weighing p-toluenesulfonic acid monohydrate (123mg, 0.65mmol, 0.1equiv.) and completely dissolving the p-toluenesulfonic acid monohydrate in 1mL of methanol, slowly dropwise adding the p-toluenesulfonic acid solution into the round-bottom flask under the ice bath condition, and reacting for 2 hours; after the reaction is finished, adding saturated saline solution, then extracting by ethyl acetate, washing for three times, drying an organic phase by anhydrous sodium sulfate, and removing an organic solvent under reduced pressure; separating by column chromatography with petroleum ether-ethyl acetate as eluent to obtain 994.2mg of pure product, yield: 80% and the product was named OH-1-CON 3 。
2) Get OTBS-1-CON 3 (2g, 6.5mmol, 1equiv.) in a dry round-bottomed flask 1, 10mL of anhydrous toluene was added to conduct azeotropic removal of water three times, and then 5mL of toluene was added asHeating the solvent to 85 ℃ under the protection of nitrogen for 2 hours for conversion, monitoring whether the conversion is complete by using a thin layer chromatography, and then cooling the system to room temperature; taking OH-1-CON 3 (1.5 g,7.8mmol,1.2 equiv.) and 0.5 percent DBTL are added into a dry round-bottom flask 2, 10mL of anhydrous toluene is added for azeotropic dehydration for three times, then 1mL of anhydrous tetrahydrofuran is added for complete dissolution, the solution in the flask 2 is transferred into a flask 1 by a double-needle transfer technology, and the reaction is carried out at normal temperature overnight; the solvent was removed by rotary evaporation under reduced pressure and column chromatography with petroleum ether-ethyl acetate afforded the product 2.6g, yield: 85%, the dimer building block was named OTBS-11-CON 3 The structure is characterized by nuclear magnetic hydrogen spectrum, and the result is shown in figure 2, figure d).
On the other hand, in a similar manner and with the same reaction steps, other types of three dimer building blocks have been synthesized, such as:
wherein, OTBS-00-CON 3 The nuclear magnetic hydrogen spectrum of (a) is shown in fig. 2, panel b).
OTBS-10-CON 3 The nuclear magnetic hydrogen spectrum of (a) is shown in fig. 2, fig. c).
Figure BDA0002575044570000131
Example 3: the first step of synthesizing a building module for synthesizing an accurate self-degradable polymer from a molecular label end to a trigger end is to prepare a monomer building element, taking the synthesis of the monomer building element with the following side group as methyl as an example:
Figure BDA0002575044570000132
one end of monomer building element molecule is benzyl hydroxyl group, and the other end is benzamide.
The preparation method comprises the following steps: 1) Putting a raw material 4-cyano-3-methylbenzoic acid (2g, 12.4mmol and 1equiv.) into a dry round-bottom flask, then adding 5mL of anhydrous tetrahydrofuran to completely dissolve the anhydrous tetrahydrofuran, connecting a system with a liquid seal under the ice bath condition, slowly adding 24.8mL of borane tetrahydrofuran solution with the concentration of 1M into the flask, heating the system to 50 ℃ after the system is stable, and reacting for 2 hours; and (3) finishing the reaction, slowly adding methanol into the system under the ice bath condition to quench until no bubbles are generated, spinning off tetrahydrofuran, adding ethyl acetate, washing with saturated salt water for three times to obtain an organic phase anhydrous sodium sulfate, drying, performing reduced pressure rotary evaporation to remove the solvent, and performing petroleum ether-ethyl acetate column chromatography to obtain a product with reduced carboxyl, wherein the yield is 1.5 g: 80 percent.
2) Dissolving the product (1g, 6.8mmol, 1equiv.) obtained in the step 1) in a round-bottom flask by using 5mL of dichloromethane, adding 6mL of 30% hydrogen peroxide under ice bath conditions, adding tetrabutylammonium hydrogen sulfate (461mg, 1.36mmol, 0.2equiv.), slowly dropwise adding a sodium hydroxide solution with the mass concentration of 20% after the addition is finished, and reacting at normal temperature for 3 hours; after the reaction is finished, spin-drying dichloromethane, performing suction filtration to obtain a solid precipitated from the system, adding ethyl acetate, heating to dissolve, performing suction filtration while hot to remove insoluble impurities, naturally cooling the filtrate, and recrystallizing to obtain 786mg of the solid, wherein the yield is as follows: 70 percent; the monomer building element is named as OH-1-CONH 2 The structure is characterized by nuclear magnetic hydrogen spectrum, and the result is shown in figure 3, a).
On the other hand, the similar monomer building units without substituent groups on the benzene ring are represented as OH-0-CONH 2 Synthesized according to the literature (Synthesis 1980,243-244 (1980)).
The second step is to prepare the building unit of the dimer, and the specific synthetic route is as follows:
Figure BDA0002575044570000141
r or R 1 Can arbitrarily represent H or CH 3 (ii) a One end of the molecular structure is benzyl hydroxyl, and the other end is benzamide.
The preparation method comprises the following steps: 1) Get OTBS-1-CON 3 (1g,3.3mmol,1equiv.)，OH-0-CONH 2 (598.8mg, 3.96mmol, 1.2equiv.) and 0.5% DBTL are added into a round-bottom flask, then 10mL of toluene is added for azeotropic dehydration for three times, 4mL of anhydrous N-methyl pyrrolidone is added as a reaction solvent, the mixture is heated to 85 ℃ under the protection of nitrogen and reacts for 4 hours,after the reaction is finished, cooling the system to room temperature, adding ethyl acetate, washing with saturated salt water for three times, removing the high-boiling-point solvent, drying the organic phase with anhydrous sodium sulfate, performing reduced-pressure rotary evaporation on the organic solvent, and performing petroleum ether-tetrahydrofuran column chromatography to obtain a product 1.2g, wherein the yield is as follows: 80 percent.
2) Dissolving 1g of the product obtained in the step 1) by using 2mL of tetrahydrofuran, adding one tenth volume of methanol to dissolve in a round-bottom flask, completely dissolving 0.1 equivalent of p-toluenesulfonic acid monohydrate by using 200 microliters of methanol, dropwise adding the solution into the round-bottom flask, reacting for 2 hours, performing suction filtration in a system to obtain a white solid, washing the white solid with deionized water for three times, performing suction filtration, and drying to obtain 631mg of a dimer product, wherein the yield is as follows: 85%, the dimer was named OH-10-CONH 2 The structure is characterized by nuclear magnetic hydrogen spectrum, and the result is shown in figure 3, figure b).
On the other hand, other types of dimeric building blocks can be synthesized in a similar manner and by the same reaction steps, for example: OH-00-CONH 2 And OH-01-CONH 2 ，
Figure BDA0002575044570000151
Example 4: synthesis of dendritic tag molecules
The dendritic tag molecules can be synthesized from hydrophilic glycol chains or hydrophobic alkyl chains, and in order to help understand the synthesis process more clearly, the synthesis of a hydrophilic dendritic tag molecule is taken as an example for illustration. The reaction formula is as follows:
Figure BDA0002575044570000152
the preparation method comprises the following steps: 1) Methyl gallate (262.5mg, 1.43mmol, 1equiv.), ground potassium carbonate powder (1.3g, 9.4mmol, 6.7equiv.), was added to a dry round-bottom flask, then the air in the flask was replaced with nitrogen, the whole system was placed under nitrogen atmosphere, 5mL of anhydrous acetonitrile was added, and heating was refluxed for half an hour; then 4 (2.6 g,4.7mmol, 3.35equiv.) is removed by azeotropy with toluene for three times, wherein 4 is synthesized according to the literature, and then is quickly added into a round-bottom flask for heating reflux reaction overnight; acetonitrile is dried by spinning, dichloromethane is added for dissolution, then saturated brine is washed for three times, an organic phase is dried by anhydrous sodium sulfate, and dichloromethane-methanol column chromatography is carried out to obtain 1.5g of a product, wherein the yield is as follows: 80 percent.
2) 1g of the product prepared in 1) was taken in a dry round-bottom flask, then 5mL of methanol was added to completely dissolve it, the mixture was heated to reflux in an oil bath, 84.6mg of potassium hydroxide was slowly added to the round-bottom flask under reflux, the potassium hydroxide was completely dissolved, and the reaction was carried out overnight. After the reaction is completed, cooling to room temperature, then slowly adding hydrochloric acid into the system, and adjusting the pH of the system to be about 3; adding dichloromethane 20mL and saturated saline water, washing for three times, drying an organic phase by anhydrous magnesium sulfate, performing suction filtration to obtain an organic phase, performing rotary evaporation under reduced pressure to dry the organic phase to obtain an oily liquid product 890mg, wherein the yield is as follows: 90 percent. The structure is characterized by nuclear magnetic hydrogen spectrum, and the result is shown in figure 4.
3) Putting the product (1g, 0.8mmol, 1equiv.) obtained in the step 2) into a dry round-bottom flask, adding 2mL of tetrahydrofuran, adding triethylamine (89mg, 0.88mmol, 1.1equiv.) under an ice bath condition, slowly dropwise adding DPPA (242.2mg, 0.88mmol, 1.1equiv.) to react for 2 hours, spin-drying the tetrahydrofuran, adding 10mL of dichloromethane, washing with 1M hydrochloric acid water for three times, washing with a saturated sodium bicarbonate solution for three times, finally washing with a saturated saline solution for three times, drying an organic phase with anhydrous magnesium sulfate, and spin-drying a solvent to obtain the product, namely dPEG-CON 3 Directly used for the next reaction.
On the other hand, other tags of the same type of structure as the hydrophilic dendritic structure are synthesized by similar means and reaction steps, for example:
Figure BDA0002575044570000161
example 5: synthesis of precise self-degradable polymer from trigger element end to molecular label end
The synthesis of the precise self-degradation polymer adopts an iterative step-by-step growth strategy, wherein the synthesis from the Trigger end to the Tag end can be obtained by a synthesis method in the following figure, namely, isocyanate and hydroxyl are coupled, and then deprotection is carried out to realize step-by-step growth. Wherein the isocyanate precursor is benzoyl azide and can be converted into isocyanate by heating. By using different building motifs in example 2 above, corresponding matched reaction motifs can be selected for different degrees of polymerization and different sequence requirements. The synthetic diagram is as follows:
Figure BDA0002575044570000171
the synthesis of the sequence T-01001001011-dPEG was chosen to further illustrate the synthesis. The first step starts with an ultraviolet-responsive Trigger synthesized according to the literature and OTBS-10-CON 3 Reacting, and then removing the protection on hydroxyl to obtain an intermediate product, wherein the specific synthetic route is as follows:
Figure BDA0002575044570000172
the preparation method comprises the following steps: 1) Dimer obtaining OTBS-10-CON 3 (730mg, 1.61mmol, 1.2equiv.) the ultraviolet light response Trigger (1g, 1.34mmol, 1equiv.), catalytic equivalent DBTL is put into a round-bottom flask, 10mL of toluene is added for azeotropic dehydration for three times, then 5mL of anhydrous tetrahydrofuran is added for dissolution, and the mixture is heated to 85 ℃ under the protection of nitrogen for reaction overnight; after the reaction time is up, cooling the system to room temperature, adding 1mL of methanol, weighing p-toluenesulfonic acid monohydrate (107.2mg, 0.67mmol,0.5 equiv.) and adding into a round-bottom flask, stirring and reacting at normal temperature for 2 hours, after the reaction is completed, adding 20mL of ethyl acetate, washing the organic solution with saturated saline solution for three times, separating to obtain an organic phase, drying with anhydrous magnesium sulfate, performing suction filtration to obtain a filtrate, performing reduced pressure rotary evaporation to remove the solvent, performing column chromatography on petroleum ether-ethyl acetate as an eluent to separate 1.25g of a product, and obtaining the yield: 90%, and the obtained intermediate product is named as T-01-OH.
2) The dimer building unit to be coupled is changed into OTBS-00-CON 3 The Trigger in the step 1) is replaced by T-01-OH, and the T-0100-OH can be obtained according to the same reaction steps; T-010010-OH, and T-01001001011-OH were also synthesized as described above.
The second step is followed by Tag from example 4 above. The specific synthetic route is as follows:
Figure BDA0002575044570000181
the preparation method comprises the following steps: the above dPEG-CON 3 (262mg, 0.2mmol, 2equiv.) is added into a dry round-bottom flask 1, then 10mL of toluene is added for azeotropic dehydration for three times, 5mL of anhydrous toluene is added as a reaction solvent, the mixture is heated to 85 ℃ for conversion for 4 hours, and the toluene is pumped to dryness after the system is cooled to room temperature; taking T-01001001011-OH (200mg, 0.1mmol, 1equiv.), adding catalytic equivalent DBTL into a round-bottom flask 2, adding 10mL of toluene for azeotropic dehydration for three times, and adding 2mL of tetrahydrofuran for complete dissolution; transferring the solution in the flask 2 into the flask 1 by using a double-needle transfer technology, and reacting for 24 hours at normal temperature under the protection of nitrogen; after the reaction is finished, carrying out reduced pressure rotary evaporation to remove most tetrahydrofuran, then precipitating the system into methanol, centrifuging to obtain a crude product, and carrying out cyclic preparation GPC separation to obtain a pure product; and the product was named T-01001001011-dPEG. The results were characterized by nuclear magnetic hydrogen spectroscopy, gel permeation chromatography and macromolecular mass spectrometry, and the results are shown in fig. 5.
In FIG. 5, FIG. a is a GPC outflow curve of T-01001001011-dPEG, which is a symmetric and narrow-distribution curve, FIG. b is a hydrogen nuclear magnetic resonance spectrum of T-01001011-dPE, FIG. c is a macromolecular mass spectrum of T-01001001001011-dPEG, the calculated molecular weight of the target molecule is 3324.84, the actually measured molecular mass spectrum value is 3325.54, and the molecular weight difference is within 1 Dalton, and it can be seen from FIG. 5 that the final product obtained according to the above synthesis steps has a precise structure and a single molecular weight.
On the other hand, the fixed Trigger end and the Tag end are of the structures as described above, and the sequence of the self-degradation segment can also be other sequences, for example: T-00000000-dPEG, T-00001111-dPEG, T-01010101-dPEG.
Example 6: synthesis of precise self-degradable polymer from molecular label end to trigger element end
The synthesis from Tag end to Trigger end can be started by orthogonal chemical reaction, and the final Trigger in the synthetic route is selectable. By using different building motifs in example 3 above, corresponding matched reaction motifs can be selected for different degrees of polymerization and different sequence requirements. The synthetic diagram is as follows:
Figure BDA0002575044570000191
when the Tag end is benzoyl azide, one end of the building element is benzamide.
The synthesis of the sequence Tag-100011-T was chosen to further illustrate the synthesis. The specific synthetic route is as follows:
Figure BDA0002575044570000192
the preparation method comprises the following steps: phy 2 -CON 3 (1g,1.35mmol,1equiv.)，OH-10-CONH 2 (505.8mg, 1.62mmol, 1.2equiv.), adding catalytic equivalent DBTL into a round-bottom flask, then adding toluene 10mL for azeotropic dehydration for three times, adding 2 mLN-methyl pyrrolidone as a reaction solvent, and heating at 85 ℃ under the protection of nitrogen for reaction for 4 hours; adding ethyl acetate, washing with saturated salt water for three times, drying an organic phase by using anhydrous sodium sulfate, and performing column chromatography by using petroleum ether-tetrahydrofuran as an eluent to obtain 1.2g of a product, wherein the yield is as follows: 80 percent.
Figure BDA0002575044570000193
The preparation method comprises the following steps: phy is added 2 -10-CONH 2 (1,1g,1.07mmol,1equiv.)，OH-00-CON 3 (387mg, 1.3mmol, 1.2equiv.), catalytic equivalent of DBTL in dry round bottom flask 1, adding toluene 10mL azeotropically to remove water three times; phINTs (481mg, 1.29mmol,1.2equiv.) Adding toluene into the flask 2 for azeotropic dehydration for 3 times; then adding 3mL of anhydrous dichloromethane into the flask 1, transferring the solution in the flask 1 into the flask 2 by using a double-needle transfer technology, and reacting for 4 hours at room temperature under the protection of nitrogen; after dichloromethane is dried by spinning, ethyl acetate is added for dissolving, saturated saline solution is used for washing for three times, and the organic phase is dried by anhydrous sodium sulfate; the petroleum ether-tetrahydrofuran was used as eluent for column chromatography to obtain 1g of product, yield: 71 percent.
In a similar way and by the same reaction steps, the ultraviolet light responding 3, 4-dimethoxy o-nitrobenzyl alcohol is blocked to obtain the final product Phy 2 -100011-NVOC having the formula:
Figure BDA0002575044570000201
the results were characterized by nuclear magnetic hydrogen spectroscopy, gel permeation chromatography and macromolecular mass spectrometry, and the results are shown in fig. 6.
In FIG. 6, panel a is Phy 2 -a GPC outflow curve of 100011-NVOC, which is a symmetric and very narrow distribution curve; panel b is Phy 2 -nuclear magnetic resonance hydrogen spectrum of 100011-NVOC; panel c is Phy 2 -a mass spectrum of macromolecules from 100011-NVOC with calculated value of 1841.99 and actual measurement 1842.43, the difference being within 1 dalton indicating the correctness of the structure; it can be seen from FIG. 6 that the final product obtained according to the above synthesis procedure has precise structure and single molecular weight.
Example 7: the gel permeation chromatography is utilized to track the degradation process of the precise self-degradation polymer and the macromolecule mass spectrum is used to determine the degradation intermediate, so as to determine the sequence of the precise self-degradation polymer
The procedure for tracking degradation is illustrated by the precision self-degrading polymer described in example 5 as T-00000000-dPEG. T-00000000-dPEG concentration of 5mg/mL (mass concentration of substance 3.7mmol/L, solvent system tetrahydrofuran, methanol, water volume ratio 5:4, sample solution 1mL was passed through a 220nm organic filter in advance, the membrane solution was sealed in a uv quartz cuvette, irradiation was stopped after 10 minutes irradiation with a uv curing lamp with power of 36W, 50 microliters of the solution under degradation was taken out with a 50 microliter microsyringe, then 50uL of THF solution containing TFA was added, further oil pump was added, 1mL (20 times) of toluene was added to the sample vial, oil pump was added, 50 microliters of pure THF was added for dissolution, microsyringe was added with a microsyringe, GPC was used to detect the precise polymer break-off, then the degradation system was incubated at constant temperature of 25 ℃, degradation was carried out for 6 hours, 14 hours, 25 hours, GPC followed the degradation, followed the degradation process by see fig. 7, fig. 8.
Wherein, FIG. 7 is a gel permeation chromatography outflow curve chart of the compound T-00000000-dPEG following degradation, FIG. 8 is an in-situ macromolecular mass spectrum of a degradation system with degradation time of 4 hours, 10 hours and 20 hours respectively after the compound T-00000000-dPEG is triggered by ultraviolet irradiation for 10 minutes, and FIG. b is an attribution chart of each peak corresponding to the mass spectrum when the compound T-000000000000-dPEG is degraded for 4 hours in FIG. a. It can be seen from fig. 7 and fig. 8 that ultraviolet light can dissociate the polymer-terminated Trigger motif, and when the Trigger motif is decomposed from the polymer, the main chain of the polymer can spontaneously undergo a degradation process from beginning to end, and a series of intermediates with different degrees of polymerization are generated.
All degradation intermediates were determined by sampling at 4, 10, and 20 hours of degradation under the same degradation conditions, and the corresponding exact polymer sequence was read according to the difference in molecular weight between each set of peaks, see fig. 7, fig. 8.
Example 8: direct reading of accurate self-degradation polymer sequences using macromolecular tandem mass spectrometry
Taking the precise self-degradable polymer structures described in the above examples 5 and 6 as an example, the macromolecular tandem mass spectrometry is used to randomly break the polymer chain segment and detect the molecular weight of the fragment peak, thereby solving the sequence structure of the complete molecule. To better understand this sequencing approach, two different secondary mass spectra of precisely self-degrading polymers are listed, see fig. 9 and 10.
Wherein, FIG. 9 is the primary mass spectrum and corresponding secondary mass spectrum of the macromolecule of T-01001011-dPEG, FIG. 10 is Phy 2 The macromolecular primary mass spectrum and the corresponding secondary mass spectrum of-100011-NVOC, and two kinds of precise polymerization can be seen from FIG. 9 and FIG. 10The sequences of the compounds can be measured by a secondary mass spectrum, but the secondary mass spectrum in FIG. 9 is cleaner, and direct reading of the precise polymer sequence can be realized by using the mass difference between adjacent peaks; as can be seen from fig. 9, the fragments in the secondary mass spectrum are only those with the branched glycol attached, because the branched glycol has a strong complexing ability for sodium ions, so that the other complementary fragment cannot be ionized, resulting in signal shielding thereof; phy 2 No branched glycol chain exists in the-100011-NVOC molecular structure, secondary mass spectrum peaks are relatively disordered, and direct reading of sequences cannot be realized.
The above description of the embodiments is only intended to facilitate the understanding of the method of the invention and its core idea. It should be noted that, for those skilled in the art, it is possible to make various improvements and modifications to the present invention without departing from the principle of the present invention, and those improvements and modifications also fall within the scope of the claims of the present invention.

Claims (5)

1. A self-degradable polymer with controllable precise sequence has a structure shown in a formula I:
Figure FDA0003967181280000011
n is any integer of 1 to 12;
Figure FDA0003967181280000012
is composed of
Figure FDA0003967181280000013
Wherein R is 2 Is H or C1-C20 alkoxy;
R 3 is H or CH 3 ；
Figure FDA0003967181280000014
Is composed of
Figure FDA0003967181280000015
R' is
Figure FDA0003967181280000016
m is an integer of 1 to 16;
y is CH 3 ；
Figure FDA0003967181280000017
Is a structural unit;
wherein R and R 1 Independently selected from H, F or Cl;
or R is H, R 1 Selected from NO 2 Alkyl with 1 to 10 carbon atoms, alkoxy with 1 to 10 carbon atoms, br or I;
when n >1, the structures of the respective structural units may be the same or different.
2. The method for preparing the precise sequence controllable self-degradable polymer as claimed in claim 1, comprising the steps of:
s1) heating and converting an intermediate shown in a formula (1) to convert an acyl azide group into an isocyanate group to obtain a phenyl isocyanate compound;
under the action of a catalyst, reacting a phenyl isocyanate compound with the hydroxyl of a triggering element hydroxyl compound to obtain a structure shown in a formula (2):
s2) removing a protecting group of hydroxyl, namely tert-butyl dimethyl silane group to obtain a compound shown as a formula (3);
s3) heating and converting the intermediate shown in the formula (1) to convert acyl azide groups into isocyanate groups to obtain phenyl isocyanate compounds;
under the action of a catalyst, reacting a phenyl isocyanate compound with a structure shown in a formula (3);
s4) repeating the steps S2-S3) for 0-11 times;
s5) removing a protecting group of hydroxyl, namely tert-butyl dimethyl silane, reacting with a compound shown as a formula (4), and blocking a label molecule to obtain a self-degradable polymer with a controllable precise sequence;
Figure FDA0003967181280000021
wherein T is
Figure FDA0003967181280000022
Wherein R is 2 Is H or C1-C20 alkoxy;
R 3 is H or CH 3 ；
Tag is
Figure FDA0003967181280000023
R' is
Figure FDA0003967181280000031
Y is CH 3 ；
m is an integer of 1 to 16;
Figure FDA0003967181280000032
is a structural unit;
R、R 1 independently selected from H, F or Cl;
or R is H, R 1 Selected from NO 2 Alkyl with 1 to 10 carbon atoms, alkoxy with 1 to 10 carbon atoms, br or I;
the structures of the individual structural units may be the same or different.
3. The method for preparing the precise sequence controllable self-degradable polymer as claimed in claim 1, comprising the steps of:
a) Reacting acyl azide groups of the label molecules shown in the formula (4) with hydroxyl groups of the compound shown in the formula (5) under a heating condition to obtain a compound shown in the formula (6);
b) Converting an amide group of the compound shown in the formula (6) into an isocyanate group under the action of an iodine reagent, and then reacting the isocyanate group with a hydroxyl group of the compound shown in the formula (7) under the action of a normal-temperature catalyst to obtain a compound shown in the formula (8);
c) Reacting acyl azide groups of the compound shown in the formula (8) with hydroxyl groups of the compound shown in the formula (5) under a heating condition, converting amide groups of obtained products into isocyanate groups under the action of an iodine reagent, and then reacting with the hydroxyl groups of the compound shown in the formula (7) under the action of a normal-temperature catalyst;
d) Repeating the step C) for 0 to 11 times;
e) Conversion to isocyanate groups under heated conditions;
then under the action of a catalyst, reacting with a trigger element shown in a structure of a formula (9) to terminate the end, so as to obtain a self-degradable polymer with controllable precise sequence;
Figure FDA0003967181280000033
Figure FDA0003967181280000041
T-OH formula (9);
wherein T is
Figure FDA0003967181280000042
Wherein R is 2 Is H or C1-C20 alkoxy;
R 3 is H or CH 3 ；
Tag is
Figure FDA0003967181280000043
R' is
Figure FDA0003967181280000044
Y is CH 3 ；
m is an integer of 1 to 16;
Figure FDA0003967181280000045
is a structural unit;
R、R 1 independently selected from H, F or Cl;
or R is H, R 1 Selected from NO 2 Alkyl with 1 to 10 carbon atoms, alkoxy with 1 to 10 carbon atoms, F, cl, br or I;
the structures of the individual structural units may be the same or different.
4. The method according to claim 2 or 3, wherein the reaction is carried out under anhydrous conditions.
5. Use of the self-degradable polymer with controllable precise sequence of claim 1 or the self-degradable polymer with controllable precise sequence prepared by the preparation method of any one of claims 2 to 4 in a novel polymer molecule sequencing technology.
CN202010651204.7A 2020-07-08 2020-07-08 Self-degradable polymer with controllable precise sequence and preparation method and application thereof Active CN113979895B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202010651204.7A CN113979895B (en) 2020-07-08 2020-07-08 Self-degradable polymer with controllable precise sequence and preparation method and application thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202010651204.7A CN113979895B (en) 2020-07-08 2020-07-08 Self-degradable polymer with controllable precise sequence and preparation method and application thereof

Publications (2)

Publication Number Publication Date
CN113979895A CN113979895A (en) 2022-01-28
CN113979895B true CN113979895B (en) 2023-03-24

Family

ID=79731263

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202010651204.7A Active CN113979895B (en) 2020-07-08 2020-07-08 Self-degradable polymer with controllable precise sequence and preparation method and application thereof

Country Status (1)

Country Link
CN (1) CN113979895B (en)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114668860A (en) * 2022-04-11 2022-06-28 中国科学技术大学 Digital lipid nanoparticle and preparation method and application thereof
CN115850122B (en) * 2022-12-16 2024-05-17 中国科学技术大学 ROMP monomer molecule with single chiral PEG side chain, and preparation method and application thereof

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2008053479A1 (en) * 2006-10-30 2008-05-08 Ramot At Tel Aviv University Ltd. Self-immolative polymers
CN101543632A (en) * 2008-11-20 2009-09-30 湘潭大学 Anti-tumor prodrug having accurate structure and taking novel amphipathic polymer as carrier and synthetic method thereof
CN101618220A (en) * 2009-05-27 2010-01-06 湘潭大学 Anti-tumor predrug by taking novel amphipathic dendritic macromolecules as carriers based on glycerol as branching unit and preparation method thereof
CN102112620A (en) * 2008-07-29 2011-06-29 帝斯曼知识产权资产管理有限公司 Method for modifying non-starch carbohydrate material using peroxidase enzyme
CN103951803A (en) * 2014-05-05 2014-07-30 中国科学技术大学 Universal method for preparing polymeric vesicles with sensitive response
CN104685064A (en) * 2012-07-24 2015-06-03 纳特拉公司 Highly multiplex PCR methods and compositions
CN105251013A (en) * 2015-09-28 2016-01-20 湘潭大学 Degradable water-soluble antitumor polymer prodrug with redox responsiveness and preparation method thereof
CN109415353A (en) * 2016-05-24 2019-03-01 美国安进公司 Pegylation Carfilzomib compound

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2008053479A1 (en) * 2006-10-30 2008-05-08 Ramot At Tel Aviv University Ltd. Self-immolative polymers
CN102112620A (en) * 2008-07-29 2011-06-29 帝斯曼知识产权资产管理有限公司 Method for modifying non-starch carbohydrate material using peroxidase enzyme
CN101543632A (en) * 2008-11-20 2009-09-30 湘潭大学 Anti-tumor prodrug having accurate structure and taking novel amphipathic polymer as carrier and synthetic method thereof
CN101618220A (en) * 2009-05-27 2010-01-06 湘潭大学 Anti-tumor predrug by taking novel amphipathic dendritic macromolecules as carriers based on glycerol as branching unit and preparation method thereof
CN104685064A (en) * 2012-07-24 2015-06-03 纳特拉公司 Highly multiplex PCR methods and compositions
CN103951803A (en) * 2014-05-05 2014-07-30 中国科学技术大学 Universal method for preparing polymeric vesicles with sensitive response
CN105251013A (en) * 2015-09-28 2016-01-20 湘潭大学 Degradable water-soluble antitumor polymer prodrug with redox responsiveness and preparation method thereof
CN109415353A (en) * 2016-05-24 2019-03-01 美国安进公司 Pegylation Carfilzomib compound

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
Self-Immolative Dendrimers;Roey J. Amir et al;《Angew. Chem. Int. Ed.》;20031231;第42卷;4494-4499 *
Self-Immolative Polymersomes for High-Efficiency Triggered Release and Programmed Enzymatic Reactions;Shiyong Liu et al;《J. Am. Chem. Soc》;20141231;第136卷;7492-7497 *
酶触发聚合物囊泡-核交联胶束转变用于响应性抗菌剂释放;刘世勇等;《高分子学报》;20170731(第7期);1178-1190 *

Also Published As

Publication number Publication date
CN113979895A (en) 2022-01-28

Similar Documents

Publication Publication Date Title
CN113979895B (en) Self-degradable polymer with controllable precise sequence and preparation method and application thereof
CN112592376B (en) Method for preparing mannose-containing derivative for post-polymerization modification by using double-click chemistry combination
CN113004170B (en) Covalent macrocyclic polymer based on column [5] arene and preparation and application thereof
WO2004019002A2 (en) Fluorescent glycosides and methods for their use
CN107245028B (en) Biphenyl aromatic hydrocarbon, water-soluble biphenyl aromatic hydrocarbon and preparation method thereof
KR102645305B1 (en) Compound based cyanine, labeling dye, kit and contrast medium composition for biomolecule comprising the same
Desimoni et al. A soluble polymer-bound Evans’ chiral auxiliary: Synthesis, characterization and use in cycloaddition reactions
WO2008067026A2 (en) Purification of synthetic oligomers
Biewend et al. Synthesis of polymer-linked copper (i) bis (N-heterocyclic carbene) complexes of linear and chain extended architecture
CN107459505B (en) Fluorescent functional carbonate, preparation method and application thereof, and fluorescent polycarbonate prepared from fluorescent functional carbonate
CN107033331A (en) A kind of fluorescence conjugated macromolecule of side-chain amino group protonation, preparation method and applications
EP3107952B1 (en) Method for the generation of chemical libraries
CN112661673A (en) Precise sequence stimuli-responsive polymer and preparation method and application thereof
KR20000022148A (en) Conjugates of an oligonucleotide/electronic conductor polymer with a molecule of interest, and their uses
Chandrasegaran et al. Phosphorus-31 NMR study of the mechanism of activation and coupling reactions in the synthesis of oligodeoxyribonucleotides by the phosphotriester method
Kajanus et al. Synthesis of bis (phenylethynyl) arylene-linked diporphyrins designed for studies of intramolecular energy transfer
Behrendt et al. Oxetane synthesis via cyclisation of aryl sulfonate esters on polystyrene and PEG polymeric supports
CN109912528B (en) Peptide-like monomer, polymer thereof and application thereof
US10239853B1 (en) Polycyclic phosphoramidtes and their applications
Deffieux et al. Towards the selective synthesis of macrocyclic vinyl polymers of controlled size
WO2005087835A1 (en) Molecular-wire type fluorescent chiral sensor
US20230021429A1 (en) Base-labile protecting groups for stepwise polymer synthesis
CN116041225B (en) AIE fluorescent compound targeted by prostate specific membrane antigen and preparation method thereof
CN114349800B (en) Ferrocene derivative and synthetic method and application thereof
Sugimoto et al. Stepped‐up reactivity of a simple lanthanoid initiator. Polymerization of methyl methacrylate initiated with a lanthanoid alkoxide–ketene system

Legal Events

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