FUNCTIONALIZED DIBLOCK COPOLYMER AND ITS PREPARATION METHOD AND APPLICATION
20250345457 ยท 2025-11-13
Assignee
Inventors
Cpc classification
A61K49/0002
HUMAN NECESSITIES
C08F301/00
CHEMISTRY; METALLURGY
A61K49/0054
HUMAN NECESSITIES
A61K41/0061
HUMAN NECESSITIES
A61K49/0093
HUMAN NECESSITIES
A61K47/6935
HUMAN NECESSITIES
International classification
A61K47/69
HUMAN NECESSITIES
C08F301/00
CHEMISTRY; METALLURGY
Abstract
A functionalized diblock copolymer. The chemical structure of the functionalized diblock copolymer is shown in Formula II. The functionalized diblock copolymers or polymer particles can be widely used in tumor imaging, tumor treatment and other fields. It not only has good safety, realizes faster and adjustable (by changing the structure and number of functional groups) degradation and removal of polymers under acidic conditions, but also has excellent specificity and high-quality imaging effects at the target site, with high signal-to-noise ratio, clear boundaries, and long half-life. It solves the problem of fluorescence imaging technology in real-time intra-operative navigation, which has a good industrialization prospect.
Claims
1. A functionalized diblock copolymer, wherein the chemical structure of the functionalized diblock copolymer is shown in Formula II: ##STR00087## wherein in Formula II, m.sub.2=221136, n.sub.2=10500, p.sub.2=0.550, q.sub.2=0500, r.sub.2=0200; s.sub.21=110, s.sub.22=110, s.sub.23=110, s.sub.24=110; n1=14, n2=14, n3=14, n4=14; L.sub.21, L.sub.22, L.sub.23, L.sub.24 are linking groups; A.sub.2 is selected from protonatable groups; C.sub.2 is selected from fluorescent molecular groups; D.sub.2 is selected from delivery molecular groups; E.sub.2 is selected from hydrophilic/hydrophobic groups; T.sub.2 is selected from capping groups; EG.sub.2 is selected from capping groups.
2. The functionalized diblock copolymer of claim 1, wherein in Formula II, the molecular weight of the polyethylene glycol block is 1,000 to 50,000 Da, and the molecular weight of the polylactone block is 1,000 to 130,000 Da; and/or, the critical micelle concentration (CMC) of the functionalized diblock copolymer is less than 50 g/mL.
3. The functionalized diblock copolymer of claim 1, wherein in Formula II, s.sub.21=16, s.sub.22=16, s.sub.23=16, s.sub.24=16; L.sub.21, L.sub.22, L.sub.23, and L.sub.24 are independently selected from S, O, OC(O), C(O)O, SC(O), C(O), OC(S), C(S)O, SS, C(R.sub.1)N, NC(R.sub.2), C(R.sub.3)NO, ONC(R.sub.4), N(R.sub.5)C(O), C(O)N(R.sub.6), N(R.sub.7)C(S), C(S)N(R.sub.8), N(R.sub.9)C(O)N(R.sub.10), OS(O)O, OP(O)O, OP(O)N, NP(O)O, NP(O)N, wherein, R.sub.1R.sub.10 are each independently selected from H, C1-C10 alkyl groups, C3-C10 cycloalkyl groups; A.sub.2 is selected from ##STR00088## wherein R.sub.11 and R.sub.12 are each independently selected from C1-C10 alkyl groups, C2-C10 alkenyl groups, C2-C10 alkynyl groups, C3-C10 cycloalkyl groups, aryl groups and heteroaryl groups; a=1-10, and a is a positive integer; preferably, R.sub.11 is n-propyl and R.sub.12 is n-propyl, or R.sub.11 is n-propyl and R.sub.12 is n-butyl, or R.sub.11 is n-butyl and R.sub.12 is n-butyl, or R.sub.11 is ethyl and R.sub.12 is ethyl, or R.sub.11 is ethyl and R.sub.12 is n-propyl; C.sub.2 is selected from ICG, METHYLENE BLUE, CY3.5, CY5, CY5.5, CY7, CY7.5, BDY630, BDY650, BDY-TMR, Tracy 645 and Tracy 652; D.sub.2 is selected from fluorescence quenching groups and drug molecule groups, wherein the fluorescence quenching group is preferably selected from BHQ-0, BHQ-1, BHQ-2, BHQ-3, BHQ-10, QXL-670, QXL-610, QXL-570, QXL 520, QXL-490, QSY35, QSY7, QSY21, QXL 680, lowa Black RQ and lowa Black FQ; wherein the drug molecule is preferably selected from chemotherapeutic drugs, more preferably selected from 5-ALA (5-Aminolevulinic acid), nucleic acid drugs, paclitaxel, cisplatin, doxorubicin, irinotecan and SN38; E.sub.2 includes one or more hydrophilic groups and/or one or more hydrophobic groups, wherein the hydrophilic/hydrophobic group is preferably selected from H, C1-C18 alkyl group, OR.sub.11 and SR.sub.12, wherein R.sub.11R.sub.12 are each independently selected from H, C1-C18 alkyl groups, C3-C10 cycloalkyl, aryl and heteroaryl groups; preferably, the hydrophilic/hydrophobic group is selected from (CH.sub.2CH.sub.2O)n-H with n being in a range of 130, (R.sub.14)NH.sub.2 where R.sub.14 is (CH.sub.2-)n with n being in a range of 118, (R.sub.15)OH where R.sub.15 is (CH.sub.2-)n with n being in a range of 118, sugar groups including monosaccharides, disaccharides, or oligosaccharide groups with various number of hydroxyl groups; preferably, the hydrophilic/hydrophobic group is selected from cholesterol and derivatives thereof, hydrophobic vitamins including Vitamin E and Vitamin D, and zwitterionic groups; wherein when hydrophilic and hydrophobic groups are used simultaneously, the total number of all hydrophilic and hydrophobic groups is described as r.sub.2, wherein r.sub.2,A is the total number of hydrophilic groups, and r.sub.2,B is the total number of hydrophobic groups, wherein, (r.sub.2,A+r.sub.2,B)=1200, and r.sub.2,A<151; preferred structures of cholesterol and derivatives thereof, Vitamin E and Vitamin D are shown below: ##STR00089## ##STR00090## preferred structures of the zwitterionic groups are shown below: ##STR00091## the hydrophilic/hydrophobic group is preferably selected from: n-nonane group, n-octane group, n-butyl group, n-propyl group, ethyl group, methyl group, n-octadecane group, n-heptadecane group, cholecterol and derivatives thereof, hydroxyethyl group, hydroxymethyl group, hydroxypropyl group, hydroxybutyl group, zwitterionic group, combination of zwiterionic group and n-nonane group, combination of zwiterionic group and n-octane group; T.sub.2 is selected from CH.sub.3 and H; and EG.sub.2 is selected from YR.sub.13, wherein, Y is selected from O, S and N, and R.sub.13 is selected from H, C1-C20 alkyl group, C3-C10 cycloalkyl, aryl and heteroaryl groups.
4. The functionalized diblock copolymer of claim 1, wherein in Formular II, m.sub.2=221136, n.sub.2=10500, p.sub.2=0.550, q.sub.2=0, r.sub.2=0; or, in Formular II, m.sub.2=221136, n.sub.2=10500, p.sub.2=0.550, q.sub.2=0, r.sub.2=1200; or, in Formular II, m.sub.2=221136, n.sub.2=10500, p.sub.2=0.550, q.sub.2=1500, r.sub.2=0.
5. The functionalized diblock copolymer of claim 1, wherein the chemical structural formula of the functionalized diblock copolymer is shown as following: ##STR00092## wherein m.sub.2=221136, preferably m.sub.2=44226, and n.sub.2=10500, preferably n.sub.2=20200, or preferably n.sub.2=35177, p.sub.2=0.55, n1=4, n2=4; ##STR00093## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, n1=4, n2=4; ##STR00094## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, n1=4, n2=4; ##STR00095## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, n1=4, n2=4; ##STR00096## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, r.sub.2=1200, n1=4, n2=4, n4=4; ##STR00097## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, r.sub.2=1200, n1=4, n2=4, n4=4; ##STR00098## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, r.sub.2=1200, n1=4, n2=4, n4=4; ##STR00099## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, r.sub.2=1200, n1=4, n2=4, n4=4; ##STR00100## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, r.sub.2=1200, n1=4, n2=4, n4=4; ##STR00101## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, r.sub.2=1200, n1=4, n2=4, n4=4; ##STR00102## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, r.sub.2=1200, n1=4, n2=4, n4=4; ##STR00103## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, r.sub.2=1200, n1=4, n2=4, n4=4; ##STR00104## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, r.sub.2=1200, n1=4, n2=4, n4=4; ##STR00105## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, r.sub.2=1200, n1=4, n2=4, n4=4; ##STR00106## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, r.sub.2=1200, n1=4, n2=4, n4=4; ##STR00107## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, r.sub.2=1200, n1=4, n2=4, n4=4; ##STR00108## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, r.sub.2=1200, n1=4, n2=4, n4=4; ##STR00109## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, r.sub.2=1200, n1=4, n2=4, n4=4; ##STR00110## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, r.sub.2=1200, n1=4, n2=4, n4=4; ##STR00111## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, r.sub.2=1200, n1=4, n2=4, n4=4; ##STR00112## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, r.sub.2=1200, n1=4, n2=4, n4=4; ##STR00113## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, r.sub.2=1200, n1=4, n2=4, n4=4; ##STR00114## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, r.sub.2=1200, n1=4, n2=4, n4=4; ##STR00115## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, r.sub.2=1200, n1=4, n2=4, n4=4; ##STR00116## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, r.sub.2=1200, n1=4, n2=4, n4=4; ##STR00117## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, r.sub.2=1200, n1=4, n2=4, n4=4; ##STR00118## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, r.sub.2=1200, n1=4, n2=4, n4=4; ##STR00119## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, r.sub.2=1200, n1=4, n2=4, n4=4; ##STR00120## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, r.sub.2=1200, n1=4, n2=4, n4=4; ##STR00121## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, r.sub.2=1200, n1=4, n2=4, n4=4; ##STR00122## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, r.sub.2=1200, n1=4, n2=4, n4=4; ##STR00123## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, r.sub.2=1200, n1=4, n2=4, n4=4; ##STR00124## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, 2=1200, n1=4, n2=4, n4=4; ##STR00125## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, r.sub.2=1200, n1=4, n2=4, n4=4; ##STR00126## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, r.sub.2=1200, n1=4, n2=4, n4=4; ##STR00127## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, r.sub.2=1200, n1=4, n2=4, n4=4; ##STR00128## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, r.sub.2=1200, n1=4, n2=4, n4=4; ##STR00129## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, r.sub.2=1200, n1=4, n2=4, n4=4; ##STR00130## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, r.sub.2=1200, n1=4, n2=4, n4=4; ##STR00131## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, r.sub.2=1200, n1=4, n2=4, n4=4; ##STR00132## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, (r.sub.2,A+r.sub.2,B)=1200, r.sub.2,A<151, n1=4, n2=4, n3=4, n4=4; ##STR00133## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, (r.sub.2,A+r.sub.2,B)=1200, r.sub.2,A<151, n1=4, n2=4, n3=4, n4=4; ##STR00134## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, (r.sub.2,A+r.sub.2,B)=1200, r.sub.2,A<151, n1=4, n2=4, n3=4, n4=4; ##STR00135## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, (r.sub.2,A+r.sub.2,B)=1200, r.sub.2,A<151, n1=4, n2=4, n3=4, n4=4; ##STR00136## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, (r.sub.2,A+r.sub.2,B)=1200, r.sub.2,A<151, n1=4, n2=4, n3=4, n4=4; ##STR00137## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, (r.sub.2,A+r.sub.2,B)=1200, r.sub.2,A<151, n1=4, n2=4, n3=4, n4=4; ##STR00138## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, (r.sub.2,A+r.sub.2,B)=1200, r.sub.2,A<151, n1=4, n2=4, n3=4, n4=4; ##STR00139## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, (r.sub.2,A+r.sub.2,B)=1200, r.sub.2,A<151, n1=4, n2=4, n3=4, n4=4; ##STR00140## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, (r.sub.2,A+r.sub.2,B)=1200, r.sub.2,A<151, n1=4, n2=4, n3=4, n4=4; ##STR00141## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, (r.sub.2,A+r.sub.2,B)=1200, r.sub.2,A<151, n1=4, n2=4, n3=4, n4=4; ##STR00142## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, (r.sub.2,A+r.sub.2,B)=1200, r.sub.2,A<151, n1=4, n2=4, n3=4, n4=4; ##STR00143## wherein, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, (r.sub.2,A+r.sub.2,B)=1200, r.sub.2,A<151, n1=4, n2=4, n3=4, n4=4; ##STR00144## wherein, m.sub.2=44226, n.sub.2=35150, p.sub.2=0.55, q.sub.2=5150, n1=4, n2=4, n3=4; ##STR00145## wherein, m.sub.2=44226, n.sub.2=35150, p.sub.2=0.55, q.sub.2=5150, n1=4, n2=4, n3=4; ##STR00146## wherein, m.sub.2=44226, n.sub.2=35150, p.sub.2=0.55, q.sub.2=5150, n1=4, n2=4, n3=4; ##STR00147## wherein, m.sub.2=44226, n.sub.2=35150, p.sub.2=0.55, q.sub.2=5150, n1=4, n2=4, n3=4; ##STR00148## wherein, m.sub.2=44226, n.sub.2=35150, p.sub.2=0.55, q.sub.2=5150, r.sub.2=1200, n1=4, n2=4, n3=4, n4=4; ##STR00149## wherein, m.sub.2=44226, n.sub.2=35150, p.sub.2=0.55, q.sub.2=5150, r.sub.2=1200, n1=4, n2=4, n3=4, n4=4; ##STR00150## wherein, m.sub.2=44226, n.sub.2=35150, p.sub.2=0.55, q.sub.2=5150, r.sub.2=1200, n1=4, n2=4, n3=4, n4=4; ##STR00151## wherein, m.sub.2=44226, n.sub.2=35150, p.sub.2=0.55, q.sub.2=5150, r.sub.2=1200, n1=4, n2=4, n3=4, n4=4; ##STR00152## wherein, m.sub.2=44226, n.sub.2=35150, p.sub.2=0.55, q.sub.2=5150, n1=4, n2=4, n3=4; ##STR00153## wherein, m.sub.2=44226, n.sub.2=35150, p.sub.2=0.55, q.sub.2=5150, n1=4, n2=4, n3=4; ##STR00154## wherein, m.sub.2=44226, n.sub.2=35150, p.sub.2=0.55, q.sub.2=5150, n1=4, n2=4, n3=4; ##STR00155## wherein, m.sub.2=44226, n.sub.2=35150, p.sub.2=0.55, q.sub.2=5150, n1=4, n2=4, n3=4.
6. Polymer particles, prepared from the functionalized diblock copolymer according to claim 1.
7. The polymer particles of claim 6, wherein the particle size of the polymer particles is 10 to 200 nm; and/or, the polymer particles are further modified with a targeting group, and the targeting group is selected from the group consisting of monoclonal antibody fragments, small molecule targeting groups, polypeptide molecules, and nucleic acid aptamers; and/or, the targeting group is modified on at least part of the T-terminal of the functionalized diblock copolymer.
8. The functionalized diblock copolymer according to claim 1, wherein the functionalized diblock copolymer and/or the polymer particles are degradable in vivo.
9. Use of the functionalized diblock copolymer according to claim 1, or polymer particles prepared from the functionalized diblock copolymer in the preparation of an imaging probe reagent and/or a pharmaceutical preparation, wherein the imaging probe reagent and/or the pharmaceutical preparation preferably has a targeting function, wherein the imaging probe reagent and/or the pharmaceutical preparation is more preferably a targeting imaging probe.
10. A composition, comprising the functionalized diblock copolymer according to claim 1, or polymer particles prepared from the functionalized diblock copolymer.
11. A method of treating or diagnosing a tumor, wherein the method comprises: administering to an individual an effective amount of the functionalized diblock copolymer according to claim 1, or administering to an individual an effective amount of polymeric particles prepared from the functionalized diblock copolymer.
12. The method according to claim 11, wherein the functionalized diblock copolymer or the polymeric particles are administered to the individual by administration methods including bladder instillation, uterus instillation, GI track instillation, topical administration to brain in an open-skull surgery, tissue injection during breast cancer dissection surgery, topical administration during minimally invasive surgery for abdominal tumor.
13. The polymer particles according to claim 6, wherein the functionalized diblock copolymer and/or the polymer particles are degradable in vivo.
Description
DESCRIPTION OF THE DRAWINGS
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DETAILED DESCRIPTION
[0057] In order to make the purpose of the invention, technical solutions and beneficial technical effects of this invention clearer, the invention will be further described in detail below in conjunction with examples. Those skilled in the art can easily understand other advantages and effects of this invention from the content disclosed in this specification.
[0058] In this invention, diblock copolymer generally refers to a polymer having two different polymer segments (as if two blocks linked together) with different chemical compositions.
[0059] In this invention, the protonatable group generally refers to a group that can combine with a proton, that is, it can bind at least one proton. These groups usually have a lone pair of electrons, so that at least one proton can be combined with the protonatable group.
[0060] In the present invention, degradability regulating group is a type of group that can change the degradability of a compound in vivo.
[0061] In this invention, fluorescent molecular group generally refers to a type of group corresponding to fluorescent molecules. Compounds containing these groups can usually have characteristic fluorescence in the ultraviolet-visible-near infrared region, and their fluorescent properties (excitation and emission wavelengths, intensity, lifetime, polarization, etc.) can change with the nature of the environment.
[0062] In this invention, delivery molecular group usually means various molecules that can be chemically bonded to the main chain of the block copolymer through a side chain, or interact with the hydrophobic side chain groups of the block copolymer through physical force (such as charge forces, hydrogen bonding, van der Waals force, hydrophobic interaction, etc.) and can be delivered by nanoparticles formed by self-assembly of the block polymer in aqueous solution. In this invention, hydrophilic/hydrophobic group generally refers to a group with a certain degree of hydrophilicity or lipophilicity.
[0063] In this invention, alkyl usually refers to a saturated aliphatic group, which can be linear or branched. For example, C1-C20 alkyl usually refers to alkyl groups with 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 carbon atom(s). Specific alkyl groups can include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, eicosyl.
[0064] In this invention, alkenyl generally refers to an unsaturated aliphatic group with CC bond(s) (carbon-carbon double bonds, ethylenic bonds), which can be straight or branched. For example, C2-C10 alkenyl generally refers to alkenyl groups of 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms. Specific alkenyl groups may include, but are not limited to, vinyl, propenyl, butenyl, pentenyl, hexenyl, heptenyl, octenyl, nonenyl, and decenyl.
[0065] In this invention, alkynyl generally refers to an unsaturated aliphatic group with CC bond(s) (carbon-carbon triple bonds, acetylene bonds), which can be straight or branched. For example, C2-C10 alkynyl generally refers to alkynyl groups of 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms. Specific alkynyl groups may include, but are not limited to, ethynyl, propynyl, butynyl, pentynyl, hexynyl, heptynyl, octynyl, nonynyl, and decynyl.
[0066] In this invention, cycloalkyl generally refers to saturated and unsaturated (but not aromatic) cyclic hydrocarbons. For example, C3-C10 cycloalkyl generally refers to cycloalkyl groups of 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms. Specific cycloalkyl groups may include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl. The term cycloalkyl in this invention also includes saturated cycloalkyls in which optionally at least one carbon atom can be replaced by a heteroatom, which can be selected from S, N, P, and O. In addition, a monounsaturated or polyunsaturated (preferably monounsaturated) cycloalkyl group without heteroatoms in the ring should belong to the term cycloalkyl group as long as it is not an aromatic system.
[0067] In this invention, aromatic group generally refers to a ring system with at least one aromatic ring and no heteroatoms. The aromatic group may be substituted or unsubstituted. The specific substituent may be selected from C1-C6 alkyl, C1-C6 alkoxy, C3-C10 cycloalkyl, hydroxyl, halogen, etc. Specific aromatic groups may include, but are not limited to, phenyl, phenol, aniline, and the like.
[0068] In this invention, heteroaryl generally refers to a ring system having at least one aromatic ring and optionally one or more (for example, 1, 2, or 3) heteroatoms selected from nitrogen, oxygen, and sulfur. The heteroaryl group may be substituted or unsubstituted, and the specific substituent may be selected from C1-C6 alkyl, C1-C6 alkoxy, C3-C10 cycloalkyl, hydroxyl, halogen and the like. Specific heteroaryl groups may include, but are not limited to, furan, benzofuran, thiophene, benzothiophene, pyrrole, pyridine, pyrimidine, pyridazine, pyrazine, quinoline, isoquinoline, phthalazine, benzo-1, 2,5-thiadiazole, benzothiazole, indole, benzotriazole, benzodioxolane, benzodioxane, benzimidazole, carbazole, or quinazoline.
[0069] In this invention, targeting agents generally refer to agents that can specifically direct a specific compound to a desired site of action (target area), which may be in the form of polymeric particles that typically have relatively low, no, or almost no interaction with non-target tissues.
[0070] In this invention, imaging probe generally refers to a class of substances that can enhance the effect of image observation after being injected (or taken) into human tissues or organs.
[0071] In this invention, individual generally includes humans and non-human animals, such as mammals, dogs, cats, horses, sheep, pigs, cows, and the like.
[0072] After a lot of practical research, the inventor of the present invention has provided a class of functionalized diblock copolymers. These diblock copolymers can be pH-responsive and degradable under corresponding pH conditions through innovative chemical modification strategies. Therefore, it can be used as a targeting agent in various fields, and the present invention has been completed on this basis.
[0073] The first aspect of the present invention provides a functionalized diblock copolymer, the functionalized diblock copolymer having the chemical structural formula shown below:
##STR00002## [0074] within formula II, m.sub.2=221136, n.sub.2=10500, p.sub.2=0.550, q.sub.2=0500, r.sub.2=0200; s.sub.21=110, s.sub.22=110, s.sub.23=110, s.sub.24=110; [0075] n1=14, n2=14, n3=14, n4=14; [0076] L.sub.21, L.sub.22, L.sub.23, L.sub.24 are linking groups; [0077] A.sub.2 is selected from protonatable groups; [0078] C.sub.2 is selected from fluorescent molecular groups; [0079] D.sub.2 is selected from delivery molecular groups; [0080] E.sub.2 is selected from hydrophilic/hydrophobic groups; [0081] T.sub.2 is selected from capping groups; [0082] EG.sub.2 is selected from capping groups.
[0083] The compound of formula II is a diblock copolymer of polyethylene glycol-polylactone, wherein the side chain structure of the polylactone block is randomly distributed, and the general formula is represented by ran.
[0084] In the compound of formula II, L.sub.21, L.sub.22, L.sub.23, L.sub.24 are usually linking groups, which is mainly used to link the main chain of the functionalized diblock copolymer and its pendant side chains. In a specific example of this invention, L.sub.21, L.sub.22, L.sub.23, L.sub.24 can be independently selected from S, O, OC(O), C(O)O, SC(O), C(O), OC(S), C(S)O, SS, C(R.sub.1)N, NC(R.sub.2), C(R.sub.3)NO, ONC(R.sub.4), N(R.sub.5)C(O), C(O)N(R.sub.6), N(R.sub.7)C(S), C(S)N(R.sub.8), N(R.sub.9)C(O)N(R.sub.10), OS(O)O, OP(O)O, OP(O)N, NP(O)O, NP(O)N, wherein, R.sub.1R.sub.10 are each independently selected from H, C1-C10 alkyl, and C3-C10 cycloalkyl.
[0085] In another specific embodiment of the present invention, L.sub.21, L.sub.22, L.sub.23, and L.sub.24 may be independently S.
[0086] In the compound of formula II, A.sub.2 is usually a protonatable group, and this group and the block of the polymer in which the group is located are mainly used to adjust the pH response of the polymer. In a specific embodiment of this invention, A.sub.2 can be
##STR00003##
wherein, R.sub.11 and R.sub.12 are each independently selected from C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C3-C10 cycloalkyl, and aryl. In another specific embodiment of the present invention, A.sub.2 can be
##STR00004##
wherein, a=1-10, and a is a positive integer.
[0087] In another specific embodiment of the present invention, A.sub.2 can be
##STR00005##
wherein R.sub.11 is ethyl, R.sub.12 is n-propyl. In another specific embodiment of the present invention, A.sub.2 is
##STR00006##
wherein, a=1-10, and a is a positive integer.
[0088] In another specific embodiment of the present invention, A.sub.2 is
##STR00007##
wherein R.sub.11 is n-propyl, R.sub.12 is n-propyl.
[0089] In another specific embodiment of the present invention, A.sub.2 is
##STR00008##
wherein R.sub.11 is n-propyl, R.sub.12 is n-butyl.
[0090] In another specific embodiment of the present invention, A.sub.2 is
##STR00009##
wherein R.sub.11 is n-butyl, R.sub.12 is n-butyl.
[0091] In another specific embodiment of the present invention, A.sub.2 is
##STR00010##
wherein R.sub.11 is ethyl, R.sub.12 is ethyl.
[0092] In another specific embodiment of the present invention, A.sub.2 is
##STR00011##
wherein R.sub.11 is ethyl, wherein R.sub.12 is n-propyl.
[0093] In the compound of formula II, C.sub.2 is usually selected from fluorescent molecular groups, and the group and the block of the polymer in which the group is located are mainly used to introduce fluorescent molecular groups. The fluorescent molecular group may specifically include, but is not limited to, one or a combination of organic reagents, metal chelate and the like. In a specific embodiment of the present invention, C2 may include fluorescent molecules such as ICG (Indocyanine Green), METHYLENE BLUE, CY3.5, CY5, CY5.5, CY7, CY7.5, BDY630, BDY650, BDY-TMR, Tracy 645, and Tracy 652.
[0094] In another specific embodiment of the present invention, C2 may include indocyanine green (ICG), and ICG may be connected to the side chain of the block through an amide bond.
[0095] In the compound of formula II, D.sub.2 can be a delivery molecular group, and this group and the block of the polymer in which the group is located are mainly used to introduce various molecular groups that can be delivered through a block copolymer. These molecular groups may include, but are not limited to, fluorescence quenching groups, drug molecule groups (for example, photodynamic therapy precursor molecules, chemotherapeutic drug molecules, biopharmaceutical molecules, etc.) and the like. In a specific embodiment of the present invention, the fluorescence quenching group can be selected from BHQ-0, BHQ-1, BHQ-2, BHQ-3, BHQ-10, QXL-670, QXL-610, QXL-570, QXL 520, QXL-490, QSY35, QSY7, QSY21, QXL 680, Iowa Black RQ, Iowa Black FQ. In a specific embodiment of the present invention, the drug molecule group can be a group corresponding to chemotherapeutic drugs, which can specifically be nucleic acid drugs, paclitaxel, cisplatin, doxorubicin, irinotecan, SN38 and other drug molecules. In another specific embodiment of the present invention, the drug molecule group may be selected from groups corresponding to photodynamic therapy chemical drugs, and may specifically be a group corresponding to 5-ALA and its derivative structure (lipidation or fatty chaining, etc.). The specific chemical structure of the group is as follows:
##STR00012##
[0096] In the compound of formula II, E.sub.2 can be selected from a hydrophilic/hydrophobic group, and this group and the block of the polymer in which the group is located are mainly used to adjust the hydrophobicity/hydrophilicity of the hydrophobic block of the polymer. In a specific embodiment of the present invention, E.sub.2 can be selected from H, C1-C18 alkyl, OR.sub.11, SR.sub.12, wherein R.sub.11R.sub.12 are each independently selected from H, C1-C18 alkyl, C3-C10 cycloalkyl, aryl, and heteroaryl. Preferably, E.sub.2 can be selected from (CH.sub.2CH.sub.2O)n-H (n=130), (R.sub.14)NH.sub.2 where R.sub.14 is repeated methylene group (CH.sub.2)n (n=118), (R.sub.15)OH where R.sub.15 is repeated methylene group (CH.sub.2-)n (n=118), sugar groups such as monosaccharides, disaccharides, or oligosaccharide groups with various number of hydroxyl groups. Preferrably, E.sub.2 can be selected from cholesterol and its' derivatives, hydrophobic vitamins such as Vitamin E and Vitamin D, and zwitterionic groups (with exemplar structures shown below). All above-mentioned hydrophilic or hydrophobic groups can either be used alone as an E.sub.2 group, or be used in combination as an E.sub.2 group. When hydrophilic and hydrophobic groups are used simultaneously, the total number of all hydrophilic and hydrophobic groups can be described as r.sub.2 where r.sub.2,A is the total number of hydrophilic groups, and r.sub.2,B is the total number of hydrophobic groups.
[0097] The chemical structural formula of the cholesterol and cholesterol derivatives, vitamin D, and vitamin E may be one of those shown below:
##STR00013## ##STR00014##
[0098] The chemical structural formula of the zwitterionic groups may be as shown in one of the following:
##STR00015##
[0099] In another specific embodiment of the present invention, E.sub.2 may be n-nonyl.
[0100] In another specific embodiment of the present invention, E.sub.2 may be n-octyl.
[0101] In another specific embodiment of the present invention, E.sub.2 may be n-buty.
[0102] In another specific embodiment of the present invention, E.sub.2 may be n-propyl.
[0103] In another specific embodiment of this invention, E.sub.2 may be ethyl.
[0104] In another specific embodiment of the present invention, E.sub.2 may be methyl.
[0105] In another specific embodiment of this invention, E.sub.2 may be n-octadecyl.
[0106] In another specific embodiment of this invention, E.sub.2 may be n-heptadecyl.
[0107] In another specific embodiment of the present invention, E.sub.2 may be cholesterol.
[0108] In another specific embodiment of the present invention, E.sub.2 may be cholesterol derivatives.
[0109] In another specific embodiment of the present invention, E.sub.2 may be hydroxyethyl.
[0110] In another specific embodiment of the present invention, E.sub.2 may be hydroxymethyl.
[0111] In another specific embodiment of the present invention, E.sub.2 may be hydroxypropyl.
[0112] In another specific embodiment of the present invention, E.sub.2 may be hydroxybutyl.
[0113] In another specific embodiment of the present invention, E.sub.2 may be selected from zwitterionic groups.
[0114] In another specific embodiment of the present invention, E.sub.2 may include a zwitterionic group and n-nonyl.
[0115] In another specific embodiment of the present invention, E.sub.2 may include a zwitterionic group and n-octyl.
[0116] In the compound of formula II, T.sub.2 can usually be selected from end groups of PEG initiators. In a specific embodiment of the present invention, T.sub.2 can be selected from CH.sub.3, H.
[0117] In the compound of formula II, EG.sub.1 can usually be produced by different capping agents added after polymerization. In a specific embodiment of the present invention, EG.sub.2 can be YR.sub.13, wherein Y is selected from O, S, and N, and R.sub.13 is selected from H, C1-C20 alkyl, C3-C10 cycloalkyl, and aromatic group.
[0118] In another specific embodiment of the present invention, EG.sub.2 can be OH.
[0119] In the compound of formula II, the molecular weight of the polyethylene glycol (PEG) block can be 100050000 Da, 10002000 Da, 20003000 Da, 30004000 Da, 40005000 Da, 50006000 Da, 60007000 Da, 70008000 Da, 80009000 Da, 900010000 Da, 1000012000 Da, 1200014000 Da, 1400016000 Da, 1600018000 Da, 1800020000 Da, 2200024000 Da, 2400026000 Da, 2600028000 Da, 2800030000 Da, 3000032000 Da, 3200034000 Da, 3400036000 Da, 3600038000 Da, 3800040000 Da, 4000042000 Da, 4200044000 Da, 4400046000 Da, 4600048000 Da, or 4800050,000 Da, the molecular weight of polylactone (PCL) block can usually be 100050000 Da, 1000-2000 Da, 2000-3000 Da, 3000-4000 Da, 4000-5000 Da, 50006000 Da, 60007000 Da, 70008000 Da, 80009000 Da, 900010000 Da, 1000012000 Da, 1200014000 Da. 1400016000 Da, 1600018000 Da, 1800020000 Da, 2200024000 Da, 2400026000 Da, 2600028000 Da, 2800030000 Da, 3000032000 Da, 3200034000 Da, 3400036000 Da, 3600038000 Da, 3800040000 Da, 4000042000 Da, 4200044000 Da, 4400046000 Da, 4600048000 Da, 4800050000 Da, 5250055000 Da, 5750060000 Da, 6000062500 Da, 6250065000 Da, 6750070000 Da, 7250075000 Da, 7750080000 Da, 825008500 Da, 8500087500 Da, 8750090000 Da, 9000092500 Da, 9250095000 Da, 9500097500 Da, 97500100000 Da, 100000102500 Da, 102500105000 Da, 105000107500 Da, 107500110000 Da, 110000112500 Da, 112500115000 Da, 115000117500 Da, 117500120000 Da, 120000122500 Da, 122500125000 Da, 125000127500 Da, or 127500130000 Da.
[0120] In a specific embodiment of the present invention, the molecular weight of the polyethylene glycol block may be 2000 to 10000 Da, and the molecular weight of the polylactone block may generally be 4000 to 20000 Da, 2000040000 Da, or 4000060000 Da.
[0121] In the compound of formula II, m.sub.2 can be 221136, 2232, 3242, 4252, 5262, 6272, 7282, 8292, 92102, 102122, 122142, 142162, 162182, 182202, 202242, 242282, 282322, 322362, 362402, 402442, 442482, 482522, 522562, 562602, 602642, 642682, 682722, 722762, 762802, 802842, 842882, 882902, 902942, 942982, or 9821136. [0122] n.sub.2 can be 10500, 1015, 1520, 2025, 2530, 3035, 3540, 4045, 4550, 4550, 5060, 6070, 7080, 8090, 90100, 100120, 120140, 140160, 160180, 180200, 200220, 220240, 240260, 260280, 280300, 300320, 320340, 340360, 360380, 380400, 400420, 420440, 440460, 460480, or 480500. [0123] p.sub.2 can be 0.550, 0.51, 12, 24, 46, 68, 810, 1012, 1214, 1416, 1618, 1820, 2025, 2530, 3035, 3540, 4045, or 4550. [0124] q.sub.2 can be 0500, 01, 12, 24, 46, 68, 810, 1012, 1214, 1416, 1618, 1820, 2025, 2530, 3035, 3540, 4045, 4550, 4550, 5060, 6070, 7080, 8090, 90100, 100120, 120140, 140160, 160180, 180200, 200220, 220240, 240260, 260280, 280300, 300320, 320340, 340360, 360380, 380400, 400420, 420440, 440460, 460480, or 480500. [0125] r.sub.2 can be 0200, 01, 12, 23, 34, 45, 67, 67, 78, 89, 910, 1012, 1214, 1416, 1618, 1820, 2025, 2530, 3035, 3540, 4045, 4550, 5060, 6070, 7080, 8090, 90100, 100120, 120140, 140160, 160180, or 180200. [0126] r.sub.2 can be the total number of all hydrophilic and hydrophobic groups, r.sub.2,A can be the total number of hydrophilic groups, and r.sub.2,B can be the total number of hydrophobic groups. Accordingly, (r.sub.2,A+r.sub.2,B)=1200, 12, 23, 34, 45, 67, 67, 78, 89, 910, 1012, 1214, 1416, 1618, 1820, 2025, 2530, 3035, 3540, 4045, 4550, 5060, 6070, 7080, 8090, 90100, 100120, 120140, 140160, 160180, or 180200; 12,A can be <151, 12, 23, 34, 45, 67, 67, 78, 89, 910, 1012, 1214, 1416, 1618, 18-20, 2025, 2530, 3035, 3540, 4045, 4550, 5060, 6070, 7080, 8090, 90100, 100120, 120140, or 140150. [0127] s.sub.21 can be 110, 12, 23, 34, 45, 67, 67, 78, 89, 910. [0128] s.sub.22 can be 110, 12, 23, 34, 45, 67, 67, 78, 89, 910. [0129] s.sub.23 can be 110, 12, 23, 34, 45, 67, 67, 78, 89, 910. [0130] s.sub.24 can be 110, 12, 23, 34, 45, 67, 67, 78, 89, 910. [0131] n1 can be 14, 12, 23, 34; [0132] n2 can be 14, 12, 23, 34; [0133] n3 can be 14, 12, 23, 34; [0134] n4 can be 14, 12, 23, 34; [0135] s.sub.21 can be 110, 12, 23, 34, 45, 67, 67, 78, 89, 910. [0136] s.sub.22 can be 110, 12, 23, 34, 45, 67, 67, 78, 89, 910. $23 can be 110, 12, 23, 34, 45, 67, 67, 78, 89, 910. [0137] s.sub.24 can be 110, 12, 23, 34, 45, 67, 67, 78, 89, 910.
[0138] In a specific embodiment of the present invention, in Formula II, m.sub.2=221136, n.sub.2=10500, p.sub.2=0.550, q.sub.2=0, r.sub.2=0. For products prepared by these polymers (for example, polymer particles, as shown in the data of
[0139] In a preferred embodiment of the present invention, the chemical structural formula of the functionalized diblock copolymer is shown in one of the following:
##STR00016##
[0140] In another preferred embodiment of the present invention, m.sub.2=221136, preferably 44226, n.sub.2=10500, preferably 20200, more preferably 35177, p.sub.2=0.55, n1=4, n2=4.
[0141] In another preferred embodiment of this invention, m.sub.2=44226, n.sub.2=35177, p.sub.2=0.55, n1=4, n2=4. In a specific embodiment of this invention, in formula II, m.sub.2=221136, n.sub.2=10500, p.sub.2=0.550, q.sub.2=0, r.sub.2=1200. For the products prepared by these polymers (for example, polymer particles), the fluorescent molecules distributed in the hydrophobic core do not emit light under certain excitation conditions (for example, in the case of near infrared ray as the excitation light source) due to the FRET effect. The addition of hydrophilic/hydrophobic groups (i.e., E.sub.2 groups) increases the stability of polymer particles, enhances the FRET effect of polymer particles (more complete fluorescence quenching), and changes the acidity sensitivity of polymer particles. After being administered to an individual, the polymer particles can be enriched at the target site (for example, tumor site) through passive targeting by EPR (or other tissue uptake methods). Since the target site has a special pH environment (for example, an acidic environment), the protonatable group (i.e., the A.sub.2 group) can be protonated in this pH range. The charge repulsion generated by its protonation and the increase in polymer solubility drive the disintegration of polymer particles. The FRET effect of the fluorophore on the dispersed polymer segment is reduced or even completely eliminated. The polymer molecules in the dispersed state enriched in the target site can emit fluorescence under certain excitation conditions (for example, in the case of near infrared ray as the excitation light source).
[0142] In a preferred embodiment of the present invention, the chemical structural formula of the functionalized diblock copolymer is as follows:
##STR00017## ##STR00018## ##STR00019## ##STR00020## ##STR00021## ##STR00022## ##STR00023## ##STR00024##
##STR00025## ##STR00026## ##STR00027##
[0143] In another preferred embodiment of the present invention, m.sub.2=221136, n.sub.2=10500, p.sub.2=0.550, q.sub.2=0, r.sub.2=1200, n1=4, n2=4, n4=4.
[0144] In a preferred embodiment of the present invention, the chemical structural formula of the functionalized diblock copolymer is as follows:
##STR00028## ##STR00029## ##STR00030## ##STR00031##
[0145] In another preferred embodiment of the present invention, m.sub.2=221136, n.sub.2=10500, p.sub.2=0.550, q.sub.2=0, (r.sub.2,A+r.sub.2,B)=1200, r.sub.2,A<151, n1=4, n2=4, n3=4, n4=4. In another preferred embodiment of the present invention, m.sub.2=44226, n.sub.2=50177, p.sub.2=0.55, r.sub.2=1200, n1=4, n2=4, n3=4.
[0146] In a specific embodiment of the present invention, in Formula II, m.sub.2=221136, n.sub.2=10500, p.sub.2=0.550, q.sub.2=1500, r.sub.2=0. For the products obtained from these polymers (for example, polymer particles), the fluorescent molecules distributed in the hydrophobic core do not emit light under certain excitation conditions (for example, in the case of near infrared ray as the excitation light source) due to the FRET effect. The delivery molecule group (i.e., the D.sub.2 group) is connected to the main chain of the functional diblock polymer via side chain branches. After being administered to an individual, it can be enriched at the target site (for example, tumor site) through passive targeting by EPR (or other tissue uptake methods). Because the target site has a special pH environment (for example, an acidic environment), the protonatable group (i.e., the A.sub.2 group) can be protonated in this pH range, and the charge repulsion generated by its protonation and the increase in polymer solubility drive the disintegration of polymer particles, and the FRET effect of the fluorophore on the dispersed single polymer segment is reduced or even completely eliminated. The polymer molecules in the dispersed state enriched in the target site can emit fluorescence under certain excitation conditions (for example, in the case of near-infrared ray as the excitation light source). In addition to the fluorescent molecular groups carried by the polymer particles, the delivery molecular groups attached to the side chains can be cleaved from the polymer (similar to pro-drug concept) via hydrolysis of linking moiety of the drug to the polymer (or alternatively via the hydrolysis of polymer backbone), under specific pH conditions at the target site. These molecules can play a corresponding role at the target site. For example, the delivery molecular group can be the group corresponding to 5-ALA, which can provide 5-ALA molecules after hydrolysis. 5-ALA can be efficiently enriched inside cancer cells with accelerated metabolism within a few hours and complete biosynthesis to form Protoporphyrin (Protoporphyrin can reside or be trapped inside the cancer cells for a longer period of time after entering the cancer cells because its metabolic process is blocked in cancer cells). At this time, fluoresce can be efficiently emitted under the irradiation of near-infrared or 400 nm excitation light. Together with ICG fluorescent molecules (excited under 780 nm), the effect of fluorescence image enhancement or boundary confirmation or cancer confirmation at the tumor site can be realized by separate fluorescence excitation under dual wavelengths. Moreover, 5-ALA is an approved precursor of photodynamic therapy drugs. In this embodiment, we creatively introduce and deliver 5-ALA, which not only enhances the effect of tumor-specific imaging, but also performs photodynamic therapy at tumor sites at the same time. In addition to the fluorescent molecular groups carried by the polymer particles, the insoluble anticancer drugs connected to the side chains form a good water-soluble, safe and stable pharmaceutical injection preparation as polymer/drug nano-particle solution. On the one hand, this pharmaceutical preparation greatly increases the solubility of hydrophobic drugs in the blood and reduces their direct contact with the blood, which improves the stability of the drug in the body, reduces the toxic and side effects of the drug, and retains the high anti-tumor activity characteristics of the drugs. After the polymer particle is disintegrated, the delivery molecular groups attached to the side chains can be cleaved from the polymer (similar to pro-drug concept) via hydrolysis of linking moiety of the drug to the polymer (or alternatively via the hydrolysis of polymer backbone), under specific pH conditions at the target site. These molecules can play a corresponding role at the target site. For example, the delivery molecule group can be the group corresponding to SN-38, which can provide SN-38 after hydrolysis of the linking moiety to the polymer, which overcomes the shortcomings of conventional hydrophobic antitumor drug delivery systems such as low drug loading capacity and strong side effects, thus improving drug safety and achieving the effect of killing cancer cells. In addition, the side chains can also be chemically connected to nucleic acid drugs or deliver nucleic acid drugs through physical action, forming a nano-formulation of nucleic acid drugs, which can significantly improve the in vivo stability of nucleic acid drugs. After the polymer particle is disintegrated, the delivery molecular groups attached to the side chains can be cleaved from the polymer (similar to pro-drug concept) via hydrolysis of linking moiety of the drug to the polymer (or alternatively via the hydrolysis of polymer backbone), under specific pH conditions at the target site. Or the drug could be released (corresponding to physical interaction/complex delivery) into the corresponding nucleic acid drug molecule under specific pH conditions at the target site, to exert drug effects at the focal site.
[0147] In a preferred embodiment of the present invention, the chemical structural formula of the functionalized diblock copolymer is as follows:
##STR00032## ##STR00033##
[0148] In another preferred embodiment of the present invention, m.sub.2=221136, n.sub.2=10500, p.sub.2=0.550, q.sub.2=1500, r.sub.2=0, n1=4, n2=4, n3=4.
[0149] In another preferred embodiment of the present invention, m.sub.2=44226, n.sub.2=50150, p.sub.2=0.55, q.sub.2=5150, n1=4, n2=4, n3=4.
[0150] In a specific embodiment of the present invention, in Formula II, m.sub.2=44226, n.sub.2=50150, p.sub.2=0.55, q.sub.2=5150, r.sub.2=1200, n1=4, n2=4, n3=4, n4=4. The addition of hydrophilic/hydrophobic groups (i.e., E.sub.2 groups) increases the stability of the polymer particles, facilitates the improvement of the blood stability and also adjusts the acidity sensitivity of the polymer particles. After being administered to an individual, the polymer particles can be enriched at the target site (for example, tumor site) through passive targeting by EPR (or other tissue uptake methods). Because the target site has a special pH environment (for example, an acidic environment), the protonatable group (i.e., the A.sub.2 group) can be protonated in this pH range, and the charge repulsion generated by the protonation and the increase in polymer solubility drive the disintegration of polymer particles, and the FRET effect of the fluorophore on the dispersed single polymer segment is reduced or even completely eliminated. The polymer molecules in the dispersed state enriched in the target site can emit fluorescence under certain excitation conditions (for example, in the case of near-infrared ray as the excitation light source). In addition to the fluorescent molecular groups carried by the polymer particles, the insoluble anticancer drugs connected to the side chains form a good water-soluble, safe and stable pharmaceutical injection preparation. On the one hand, this pharmaceutical preparation greatly increases the solubility of hydrophobic drugs in the blood and reduces their direct contact with the blood, which improves the stability of the drug in the body, reduces the toxic and side effects of the drugs, and retains the high anti-tumor activity characteristics of the drugs. After the polymer particle is disintegrated, the delivery molecular groups attached to the side chains can be cleaved from the polymer (similar to pro-drug concept) via hydrolysis of linking moiety of the drug to the polymer (or alternatively via the hydrolysis of polymer backbone), under specific pH conditions at the target site. These molecules can play a corresponding role at the target site. For example, the delivery molecule group can be the group corresponding to SN-38, which can provide SN-38 after hydrolysis, which overcomes the shortcomings of conventional hydrophobic antitumor drug delivery systems such as low drug loading capacity and strong side effects, thus improving drug safety and achieving the effect of killing cancer cells. In addition, the side chains can also be chemically connected to nucleic acid drugs or deliver nucleic acid drugs through physical action, forming a nano-formulation of nucleic acid drugs, which can significantly improve the in vivo stability of nucleic acid drugs. After the polymer particle is disintegrated, the delivery molecular groups attached to the side chains can be cleaved from the polymer (similar to pro-drug concept) via hydrolysis of linking moiety of the drug to the polymer (or alternatively via the hydrolysis of polymer backbone), under specific pH conditions at the target site, to exert drug effects at the focal site. Or the drug could be released (corresponding to physical interaction/complex delivery) into the corresponding nucleic acid drug molecule under specific pH conditions at the target site, to exert drug effects at the focal site.
##STR00034##
[0151] In another preferred embodiment of this invention, m.sub.2=44226, n.sub.2=50150, p.sub.2=0.55, q.sub.2=5150, r.sub.2=1200, n1=4, n2=4, n3=4, n4=4.
[0152] The functionalized diblock copolymer provided in this invention usually has a low critical micelle concentration (CMC), thereby reducing the difficulty of preparing polymer self-assembled particles, thereby ensuring that the prepared polymer particles have good stability in solution and blood. For example, the critical micelle concentration (CMC) of the functionalized diblock copolymer may be <50 g/mL, <45 g/mL, <40 g/mL, <35 g/mL, <30 g/mL, <25 g/mL, <20 g/mL, <16 g/mL, <14 g/mL, <12 g/mL, <10 g/mL, 9 g/mL, 8 g/mL, 7 g/mL, 6 g/mL, 5 g/mL, 4 g/mL, or smaller critical micelle concentration.
[0153] The second aspect of the present invention provides a polymer particle prepared from the functionalized diblock copolymer provided in the first aspect of the present invention. The functionalized diblock copolymers described above can be used to form polymer particles. The fluorescent molecules distributed in the hydrophobic core of the polymer particles do not emit light under certain excitation conditions (for example, in the case of near infrared as the excitation light source) due to the FRET effect. After being administered to an individual, the polymer particles can be enriched at the target site (for example, tumor site) through passive targeting by EPR (or other tissue uptake methods). Because the target site has a special pH environment (for example, an acidic environment), the protonatable group can be protonated in this pH environment, and the charge repulsion generated by the protonation and the increase in polymer solubility drive the disintegration of polymer particles, and the FRET effect of the fluorophore on the dispersed single polymer segment is reduced or even completely eliminated, allowing the polymer molecules in the dispersed state enriched in the target site to emit fluorescence under certain excitation conditions (for example, in the case of near-infrared ray as the excitation light source). For example, the aforementioned pH environment can be 6.5-6.8, which can correspond to the interstitial fluid of tumor tissue, and at least part of the polymer particles can reach the target site and be in the interstitial fluid of the cells; for another example, the aforementioned pH environment can also be 4.5-6.5, which correspond to endosomes or lysosomes in tumor cells (or normal cells), and at least part of the polymer particles can interact with cells at the target site (for example, tumor cells) and enter into the cells through the endocytosis mechanism, thus reaching the above pH environment. The polymer particles prepared by the functionalized diblock copolymer provided in this invention can be sufficiently diffused at the target site to achieve a clear fluorescence margin, and the functionalized diblock copolymer and/or polymer particles are bio-degradable in vivo. After being administered to an individual, polymer particles or nano-particles may be up-taken by the body's immune system (mainly macrophages, etc.) and then degraded although PEG cannot be completely degraded in the body, PEG molecules with a molecular weight of less than 40,000 Da (for example, Roche's long-acting interferon, PEGASYS, has been approved for safe clinical use for more than ten years, and its molecular weight is 40,000 Da) can be effectively eliminated by the kidneys after circulating in the body); PCL block can be eliminated by the kidneys after its molecular weight is gradually reduced by hydrolysis). The immune system uptake causes short circulation and poor accumulation to the tumor site through the EPR effect cycle. The polymer particles targeted to the target site through the EPR effect are disintegrated into free functionalized diblock copolymer molecules, which, under the pH conditions of the target site and the presence of a variety of enzymes, can be degraded into PEG (which can be cleared by the kidney after circulation) and degradable block (PCL) polymers with gradually smaller molecular weights (which can be subsequently metabolized by circulation and partially cleared by the kidneys). These degradation pathways can improve the safety of the drug system for imaging probe applications or drug delivery system applications that are implemented (dose) in a single or multiple doses. Imaging observation results of live animals are shown in
[0154] The polymer particles provided in this invention can be nano-sized. For example, the particle size of the polymer particles can be 10200 nm, 1020 nm, 2030 nm, 3040 nm, 4060 nm, 6080 nm, 80 nm100 nm, 100120 nm, 120140 nm, 140160 nm, 160180 nm, or 180200 nm.
[0155] In the polymer particles provided in the present invention, the polymer particles can also be modified with targeting groups, and these targeting groups can usually be modified on the surface of the polymer particles. Suitable methods for modifying the targeting group on the polymer particles should be known to those skilled in the art. For example, in general, the targeting group can be attached to the T end of the molecular structure of the functionalized diblock copolymer. These targeting groups can usually increase the efficiency of targeting nanoparticles to tumors based on the EPR effect (or other tissue uptake methods). These targeting groups can include but are not limited to various functional molecules such as (monoclonal) antibody fragments (for example, Fab, etc.), small molecule targeting groups (for example, folic acid, carbohydrates), polypeptide molecules (for example, cRGD, GL2P), and aptamers, and these functional molecules may have a targeting function (for example, a function of targeting tumor tissue). In a specific embodiment of the present invention, the targeting group is selected from -GalNac (N-acetylgalactosamine).
[0156] The third aspect of the present invention provides a method for preparing the polymer particles provided in the second aspect of the present invention. Based on the knowledge of the chemical structure of the functionalized diblock copolymer, a suitable method for forming polymer particles shall be known to those skilled in the art. For example, the method may include: dispersing an organic solvent including the above-mentioned functionalized diblock copolymer in water for self-assembling to provide the polymer particles; or conversely, dispersing water in an organic solvent including the functionalized diblock copolymer. In the above dispersion process, proper operations can be used to make the system fully mixed, for example, it can be carried out under ultrasonic conditions. For another example, the self-assembly process can usually be carried out by removing the organic solvent in the reaction system. The organic solvent removal method may specifically be a solvent volatilization method, an ultrafiltration method, and the like. For another example, the critical micelle concentration (CMC) of a polymer is related to the ratio of the hydrophobic block to the hydrophilic block of the polymer. The higher the ratio of the hydrophobic block, the smaller the CMC. When E1, E2, E3 are long-chain hydrophobic side chains, their content is inversely proportional to the value of CMC; when E1, E2, E3 are hydrophilic side chains, their content is directly proportional to the value of CMC. For another example, the particle size of polymer particles can usually be adjusted by an extrusion instrument or microfluidic devices (homogenizers, NanoAssemblr, etc.).
[0157] The fourth aspect of the invention provides the uses of the functionalized diblock copolymer provided in the first aspect of the invention or the polymer particles provided in the second aspect of the invention in the preparation of pharmaceutical preparations and/or reagents. Polymer nanoparticles formed as a drug delivery system allow the delivery of drugs or imaging probe molecules using polymer particles as carriers. As mentioned above, the products (for example, polymer particles) prepared by the functionalized diblock copolymers provided in this invention have a passive targeting (enriched at the tumor sites through the general EPR effect of nanoparticles) or active targeting (enriched at the tumor sites by specific binding of nanoparticle surface-modified targeting groups to tumor surface-specific receptors) function. After administration to the individual, because the target site has a special pH environment (for example, acidic environment), the protonatable group can be protonated in this pH range. The charge repulsion generated by the protonation and the increase in polymer solubility drive the disintegration of polymer particles, and the FRET effect of the fluorophore on the dispersed single polymer segment is reduced or even completely eliminated, allowing the polymer molecules in the dispersed state enriched in the target site to emit fluorescence under certain excitation conditions (for example, in the case of near-infrared ray as the excitation light source). In this case, the target site (for example, the tumor site) can emit light specifically, which can be used for targeted imaging probing. In addition to imaging probe applications, these polymer particles can be used to prepare targeting agents. In a specific embodiment of this invention, the above polymer particles can be used to prepare polymer particle-based drug delivery systems to deliver various drug molecules.
[0158] In the pharmaceutical preparations or reagents provided by the present invention, polymer particles can usually be used as carriers to deliver drugs or imaging probe molecules. The functionalized diblock copolymer can be used as a single active ingredient or can be combined with other active components to collectively form the active ingredient for the aforementioned uses.
[0159] The fifth aspect of the present invention provides a composition comprising the functionalized diblock copolymer provided in the first aspect of the present invention or the polymer particles provided in the second aspect of the present invention. As mentioned above, the aforementioned composition may be a targeting agent, and in a specific embodiment of the present invention, the aforementioned composition may be an imaging probe.
[0160] The composition provided in this invention may also include at least one pharmaceutically acceptable carrier, which usually refers to a carrier for administration, which does not induce the production of antibodies harmful to the individual receiving the composition, and are not excessively toxic after administration. These carriers are well-known to those skilled in the art. For example, Remington's Pharmaceutical Sciences (Mack Pub. Co., N. J. 1991) discloses related content about pharmaceutically acceptable carriers. Specifically, the carrier may be one or more of saline, buffer, glucose, water, glycerol, ethanol, and adjuvant.
[0161] In the composition provided in this invention, the functionalized diblock copolymer may serve as a single active ingredient, or can be combined with other active components for joint use. The other active components can be various other drugs and/or agents, which can usually act on the target site together with the above-mentioned functionalized diblock copolymer. The content of the active ingredient in the composition is usually a safe and effective amount, and the safe and effective amount should be adjustable for those skilled in the art. For example, the dosage of the active ingredient usually depends on the body weight of the subject to whom it is administered, the type of application, and the condition and severity of the disease.
[0162] The composition provided in this invention can be adapted to any form of administration, and can be parenteral administration, for example, it can be pulmonary, nasal, rectal and/or intravenous injection (as shown in
[0163] The sixth aspect of the invention provides a method of treatment or diagnosis, comprising: administering to an individual an effective amount of the functionalized diblock copolymer provided in the first aspect of the invention, or the polymer particles provided in the second aspect of the invention, or the composition provided by the fifth aspect of the present invention. The effective amount generally refers to an amount that can achieve the desired effect after a proper administration period, for example, imaging, treatment of diseases, etc. The above-mentioned are pH-responsive and can be degraded under corresponding pH conditions. Chemical modifications on the functionalized diblock copolymers can also bring synergistic effect from co-delivered molecules, which are bonded to polymer molecules through degradable/cleavable chemical bonds, and can be combined with a unique end group (targeting group, a group that can improve the immunogenicity of the system) to become a unique block copolymer-delivery bonded complex. In a specific embodiment of the present invention, after use, better intraoperative tumor boundary discrimination, more precise removal of tumor lesions and metastatic tissue, and intraoperative imaging can be achieved. The delivery molecule (ie drug) can better kill cancer cells, reduce the recurrence rate, and improve the postoperative survival rate of patients.
[0164] The functionalized diblock copolymers or polymer particles provided in this invention can significantly improve the safety of tumor imaging probe reagents and/or tumor drug preparations (tumor imaging probe reagents are mostly for single use; and tumor drugs are usually administered multiple times). For the diblock copolymer provided by the present invention (the compound of formula II, or PEG-PCL diblock copolymer), PEG can be safely removed from the human body (ADEGEN, ONCASPAR, etc., are clinically approved to use PEG with a molecular weight of 5K in multi-site modified therapeutic enzymes; biological macromolecules such as interferon, granulocyte colony stimulating factor, and antibody Fab fragments modified with 12-40K PEG have been safely used in clinical practice for more than ten years), another block PCL can be gradually degraded under physiological conditions (hydrolysis; enzyme), and the imaging data of the living body proves that the PCL system can be metabolized and eliminated in experiments made on the living.
[0165] The functionalized diblock copolymers or polymer particles provided in this invention can achieve high-quality imaging with tumor imaging probe reagents specific to solid tumor sites, and can be sensitive to pH changes at the tumor site (fluorescence signal changes A pH10-90% only need about 0.2-0.3 pH unit), with high signal-to-noise ratio (as shown in
[0166] The functionalized diblock copolymers or polymer particles provided in this invention enable in vivo labeling of cancerous lymph nodes after administration, as shown in
[0167] The functionalized diblock copolymers, polymer particles, or compositions provided in this invention can be conveniently administered locally, for example, bladder instillation (as shown in
[0168] The functionalized diblock copolymers or polymer particles provided in this invention can introduce, based on the feature that the nanoparticles can accumulate sufficiently into the solid tumor microenvironment, precursor molecules (e.g., precursor molecules for photodynamic therapeutics, more specifically precursor molecules of 5-ALA) to the polymers, where these precursor molecules can be cleaved and release to the tumor microenvironment (weak acids, tumor microenvironment-specific proteases, etc.). The side chains can be cleaved from the polymer backbone and converted to the clinically approved drug molecules (e.g., 5-ALA, etc.), enabling intraoperative image enhancement of tumor sites. At the same time as the implementation of imaging, the designed imaging probe reagent utilizes the light source of intraoperative imaging to realize the photodynamic therapy of tumor tissue during tumor resection surgery, and reduce the damage of other photodynamic therapy on normal tissue, kill the uncut cancer tissue in the process of resection of the tumor tissue, reduce postoperative recurrence and prolong survival time.
[0169] In summary, the functionalized diblock copolymers or polymer particles provided in this invention can be widely used in tumor imaging, tumor therapy and other fields. It not only has good safety, realizes faster and adjustable degradation and removal of polymers (by changing the structure and number of functional groups) under acidic conditions, but also has excellent specific and high-quality imaging effects at the target site, with high signal-to-noise ratio, clear boundaries, long half-life, etc., which solves the problem of fluorescence imaging technology in real-time intraoperative navigation, and thus has a good industrialization prospect.
[0170] The following embodiments further illustrate the invention, but do not limit the scope of this invention.
[0171] The reaction route of the preparation method of the compound of formula II series in the embodiment is as follows:
##STR00035##
Example 1
1 Synthetic Schematic Diagram of PCL Fluorescent Probe
##STR00036##
2.1 Synthesis of Monomers and Side Chain Molecules
2.1.1 Synthesis of 3-Bromocaprolactone
##STR00037##
Step 1: Synthesis of 2-bromocyclohexanone (IB004-022-01)
[0172] P-TsOH (P-toluenesulfonic acid, 9.5 g, 0.05 mol) and NBS (N-Bromosuccinimide, 106.8 g, 0.6 mol) were dissolved in 400 ml dichloromethane (DCM), cooled to 0 C. in an ice bath, and 100 ml cyclohexanone (49 g, 0.5 mol) in DCM solution was slowly added while stirring. After the addition of the cyclohexanone in DCM solution, the reaction system was heated to reflux, and the reaction was carried out overnight. DCM was concentrated, 100 ml H.sub.2O was added, and DCM was added for extraction three times (50 ml3). The organic phases were combined, and the solution was washed through saturated NaHCO.sub.3 solution (50 ml1), H.sub.2O (50 ml1), and saturated NaCl solution (50 ml1). The combined organic phase was dried with Na.sub.2SO.sub.4, filtered, and concentrated. The resulting crude product was distilled under reduced pressure (75 C./0.4 torr) to obtain 46 g of colorless transparent liquid with a yield of 51.6%.
Step 2: Synthesis of 3-bromocaprolactone (IB004-026-01)
[0173] 2-Bromocyclohexanone (35.2 g, 0.2 mol) was dissolved in 300 ml DCM, and m-CPBA (m-chloroperoxybenzoic acid, 34.6 g, 0.26 mol) was added in batches. After the addition, the solution were stirred at room temperature overnight. Then the solid was filtered off, and 100 ml saturated Na.sub.2S.sub.2O.sub.3 solution was added to wash the solution. The organic phase was separated, washed with saturated NaHCO.sub.3 (100 ml1) solution, H.sub.2O (100 ml1), and saturated NaCl (100 ml1) solution in sequence, dried with anhydrous Na.sub.2SO.sub.4, filtered, and concentrated to obtain the crude product, which was then distilled under reduced pressure (85 C./0.4 torr). 12.4 g of colorless and transparent liquid was obtained with a yield of 32.3%. .sup.1H NMR (400 MHz, CDCl.sub.3) 4.86 (dd, J=5.4, 4.1 Hz, 1H), 4.74 (ddd, J=12.7, 9.4, 1.2 Hz, 1H), 4.31 (ddt, J=12.8, 6.8, 1.4 Hz, 1H), 2.20-1.77 (m, 6H).
2.1.2 Synthesis of TEPr:
##STR00038##
[0174] N-ethyl n-propylamine (34.8 g, 0.4 mol) and 500 ml dichloromethane were added to a 1 L three-necked flask sequentially. The system was replaced with N.sub.2 three times, and then ethylene sulfide (48 g, 0.8 mol) was slowly added dropwise to the above solution. After the dropping is completed, the reaction system is stirred and reacted overnight at room temperature. Then the reaction was terminated, the organic solvent was concentrated, and the finally obtained concentrate was distilled under reduced pressure (0.2 torr, 38 C.) to obtain 24 g of product, which was a colorless and transparent liquid with a yield of 40.8%. .sup.1H NMR (400 MHz, CDCl.sub.3) 4.81 (d, J=8.5 Hz, 4H), 2.67-2.46 (m, 6H), 2.37 (dd, J=8.6, 6.5 Hz, 2H), 1.51-1.37 (m, 2H), 1.00 (t, J=7.1 Hz, 3H), 0.87 (t, J=7.3 Hz, 3H).
2.1.3 Synthesis of TPrPr:
##STR00039##
[0175] Di-n-propylamine (40.4 g, 0.4 mol) and 500 ml dichloromethane were added to a 1 L three-necked flask sequentially. The system was replaced with nitrogen three times, and then ethylene sulfide (48 g, 0.8 mol) was slowly added dropwise to the above solution. After the dropping, the reaction system was stirred and reacted overnight at room temperature. Then the reaction was terminated, the organic solvent was concentrated, and the final concentrate was distilled under reduced pressure (0.2 torr, 42 C.) to obtain 21 g of product, which was a colorless and transparent liquid with a yield of 32.6%. .sup.1H NMR (400 MHZ, CDCl.sub.3) 2.69-2.54 (m, 4H), 2.39 (dd, J=8.5, 6.6 Hz, 4H), 1.46 (h, J=7.4 Hz, 4H), 0.89 (t, J=7.4 Hz, 6H).
2.1.4 Synthesis of 5-ALA Side Chain
##STR00040##
Step 1: Synthesis of 6-Tritylsulfanyl-hexan-1-ol (IB004-045-01)
[0176] Triphenylmethyl mercaptan (8.29 g, 0.03 mol) was dissolved in 30 ml EtOH and 30 ml water, then K.sub.2CO.sub.3 (4.14 g, 0.03 mol) was added. The reaction is reacted under argon, and the solution was stirred at room temperature for 30 min. Bromohexanol (5.43 g, 0.03 mol) was added and the temperature was raised to 80 C. Then the reaction was stirred overnight. The reaction was stopped and the solution was filtered and EtOH was concentrated. 50 ml water was added and the solution was extracted with EA (Ethyl acetate, 50 ml3). The organic phases were combined and washed with water (50 ml1) and saturated NaCl (50 ml1), dried with anhydrous Na.sub.2SO.sub.4, filtered, concentrated, and pulled dry by an oil pump to obtain 10.92 g of white solid, with a yield of 96.4%, which was directly used in the next step without further purification. .sup.1H-NMR (500 MHz, Chloroform-d) 7.49-7.39 (m, 6H), 7.29 (t, J=7.7 Hz, 6H), 7.25-7.18 (m, 3H), 3.58 (t, J=6.6 Hz, 2H), 2.16 (t, J=7.3 Hz, 2H), 1.54 (s, 1H), 1.53-1.45 (m, 2H), 1.45-1.38 (m, 2H), 1.34-1.18 (m, 4H).
Step 2: Synthesis of 5-(9H-Fluoren-9-ylmethoxycarbonylamino)-4-oxo-pentanoic acid
[0177] 6-tritylsulfanyl-hexyl ester (IB004-055-01) 6-Triphenylmercaptohexan-1-ol (3.77 g, 0.01 mol) was dissolved in 30 ml tetrahydrofuran (THF), then SOCl.sub.2 (1.67 g, 0.014 mol) was added and stirred for 10 min. 5-Fmoc-5-aminolevulinic acid hydrochloride (1.67 g, 0.01 mol) was added, stirred at room temperature and reacted overnight. 50 ml of saturated NaHCO.sub.3 solution was added slowly and the resulting solution was extracted with EA (50 ml3). The organic phases were combined and washed with water (50 ml1) and saturated NaCl (50 ml1), dried with anhydrous Na.sub.2SO.sub.4, filtered, and concentrated. The crude product was separated and purified by silica gel column (EA:PE=1:25). A total of 4.17 g of product was obtained, which was a colorless and transparent oily substance with the yield of 58.7%. .sup.1H-NMR (500 MHZ, Chloroform-d) 7.89 (m, 2H), 7.73-7.65 (m, 4H), 7.49-7.39 (m, 8H), 7.29 (t, J=7.7 Hz, 6H), 7.25-7.18 (m, 3H), 4.07 (2H, br s), 3.58 (t, J=6.6 Hz, 2H), 2.87 (2H, t, J=6.5 Hz), 2.63 (2H, t, J=6.5 Hz), 2.16 (t, J=7.3 Hz, 2H), 1.53-1.45 (m, 2H), 1.45-1.38 (m, 2H), 1.34-1.18 (m, 4H).
Step 3: Synthesis of 5-(9H-Fluoren-9-ylmethoxycarbonylamino)-4-oxo-pentanoic acid 6-mercapto-hexyl ester (IB004-063-01)
[0178] 6-triphenylmercaptohexyl 5-Fmo-5-amino-4-oxovalerate (3.56 g, 5 mmol) was dissolved in 50 ml DCM, and then Et.sub.3SiH (3.41 g, 29.4 mmol) and TFA (6.7 g, 58.8 mmol) were added. The solution was stirred at room temperature for 1 h and the solvent was concentrated. And then 50 ml of water was added, and 50 ml of saturated NaHCO.sub.3 solution was added slowly, and the resulting solution was extracted with EA (50 ml3). The organic phases were combined and washed with water (50 ml1) and saturated NaCl (50 ml1), dried with anhydrous Na.sub.2SO.sub.4, filtered, concentrated. The crude product was separated and purified by silica gel column (EA:PE=1:5). A total of 1.05 g of product was obtained, which was a colorless transparent oily substance with the yield of 44.8%. .sup.1H-NMR (500 MHZ, Chloroform-d) 7.89 (m, 2H), 7.73-7.65 (m, 4H), 7.45 (m, 2H), 4.31-4.25 (m, 3H), 4.07 (m, 4H), 3.57 (t, J=6.6 Hz, 2H), 2.87 (2H, t, J=6.5 Hz), 2.63 (2H, t, J=6.5 Hz), 2.16 (t, J=7.3 Hz, 2H), 1.54 (s, 1H), 1.53-1.45 (m, 2H), 1.45-1.38 (m, 2H), 1.32-1.15 (m, 4H).
2.2 Synthesis:
##STR00041##
2.2.1 PCL38 (IB004-078-01)
[0179] In a glove box with H.sub.2O and O.sub.2 indicators less than 0.1 ppm, weigh m-PEG-5000 (100 mg, 0.02 mol) and 3-bromocaprolactone (230.4 mg, 1.2 mmol) into the polymerization reaction tube, and add 300 L toluene, finally add Sn(Oct).sub.2 (8.1 mg, 0.02 mmol), react at 100 C. for 1.5 h, then cool to room temperature, add 1 ml DCM to completely dissolve it, then precipitate in 50 ml methyl tert-butyl ether. After stirring for 10 minutes, it was filtered. The crude product was vacuum dried to obtain 195 mg of white solid polymer, with a yield of 59%. .sup.1H NMR (400 MHZ, CDCl.sub.3) 4.27-4.11 (m, 105H), 3.65 (s, 448H), 3.39 (s, 3H), 2.17-1.97 (m, 77H), 1.80-1.40 (m, 151H). Mw: 16693, Mn: 14008, PDI: 1.192.
2.2.2 PCL56 (IB004-087-01)
[0180] The synthesis and purification of PCL56 were performed according to the procedure in example 2.2.1 above (using 268.8 mg of 3-bromocaprolactone, 1.4 mmol) to obtain a total of 320.5 mg of white solid polymer with the yield of 86.9%. .sup.1H NMR (400 MHZ, CDCl.sub.3) 4.27-4.13 (m, 148H), 3.65 (s, 448H), 3.39 (s, 3H), 2.18-1.97 (m, 107H), 1.80-1.40 (m, 215H). Mw: 21833, Mn: 17678, PDI: 1.235.
2.2.3 PCL70 (IB004-088-01)
[0181] The synthesis and purification of PCL70 were performed according to the procedure in example 2.2.1 above (using 307.2 mg of 3-bromocaprolactone, 1.6 mmol) to obtain a total of 362 mg of white solid polymer with the yield of 89%. .sup.1H NMR (400 MHZ, Chloroform-d) 4.28-4.11 (m, 193H), 3.65 (s, 448H), 3.39 (s, 3H), 2.19-1.97 (m, 138H), 1.81-1.40 (m, 274H). Mw: 21365, Mn: 17609, PDI: 1.213.
2.2.4 PCL83 (IB004-094-01)
[0182] The synthesis and purification of PCL83 were performed according to the procedure in example 2.2.1 above (using 3-bromocaprolactone 345.6 mg, 1.8 mmol) to obtain a total of 415 mg of white solid polymer with a yield of 93.1%. .sup.1H NMR (400 MHZ, CDCl.sub.3) 4.26-4.12 (m, 246H), 3.66 (s, 448H), 3.39 (s, 3H), 2.18-1.97 (m, 172H), 1.80-1.40 (m, 359H). Mw: 26301, Mn: 20997, PDI: 1.253.
2.2.5 PCL100 (IB004-080-01)
[0183] The synthesis and purification of PCL100 were performed according to the procedure in example 2.2.1 above (using 3-bromocaprolactone 384 mg, 2.0 mmol) to obtain a total of 435.6 mg of white solid polymer with a yield of 90%. .sup.1H NMR (400 MHZ, CDCl.sub.3) 4.28-4.12 (m, 285H), 3.65 (s, 448H), 3.39 (s, 3H), 2.19-1.97 (m, 206H), 1.81-1.40 (m, 402H). Mw: 27118, Mn: 20108, PDI: 1.349.
2.2.6 PCL115 (IB004-114-01)
[0184] The synthesis and purification of PCL115 were performed according to the procedure in example 2.2.1 above (using 3-bromocaprolactone 442 mg, 2.3 mmol) to obtain a total of 3.78 g of white solid polymer with the yield of 96.4%. .sup.1H NMR (400 MHZ, CDCl.sub.3) 4.27-4.13 (m, 342H), 3.66 (s, 448H), 3.39 (s, 3H), 2.18-1.99 (m, 240H), 1.80-1.41 (m, 492H). Mw: 27081, Mn: 20564, PDI: 1.317.
2.2.7 PCL150 (IB004-108-01)
[0185] The synthesis and purification of PCL150 were performed according to the procedure in example 2.2.1 above (using 3-bromocaprolactone 654 mg, 3.4 mmol) to obtain a total of 742 mg of white solid polymer with a yield of 98.5%, .sup.1H NMR (400 MHZ, CDCl.sub.3) 4.29-4.14 (m, 464H), 3.67 (s, 448H), 3.40 (s, 3H), 2.19-1.99 (m, 321H), 1.81-1.42 (m, 659H). Mw: 35456, Mn: 26806, PDI: 1.323.
2.2.8 PCL180 (IB004-107-01)
[0186] The synthesis and purification of PCL180 were performed according to the procedure in example 2.2.1 above (using 3-bromocaprolactone 768 mg, 4.0 mmol) to obtain a total of 859 mg white solid polymer with a yield of 99%. .sup.1H NMR (400 MHZ, CDCl.sub.3) 4.28-4.12 (m, 560H), 3.65 (s, 448H), 3.39 (s, 3H), 2.18-1.97 (m, 389H), 1.81-1.40 (m, 795H). Mw: 36002, Mn: 25265, PDI: 1.425.
2.2.9 HO-PCL115
##STR00042##
Step 1: Bn-PCL115 (IB004-119-01)
[0187] The synthesis and purification of Bn-PCL115 were performed according to the process in example 2.2.1 above, in which m-PEG-5000 was replaced with an equimolar amount of Bn-PEG-5000 to obtain a total of 4.36 g of white solid polymer with a yield of 97.1%. .sup.1H NMR (400 MHZ, CDCl.sub.3) 7.30 (m, 5H), 5.11 (s, 2H), 4.28-4.14 (m, 342H), 3.66 (s, 448H), 2.19-2.00 (m, 240H), 1.82-1.41 (m, 492H). Mw: 29948, Mn: 24753, PDI: 1.210.
Step 2: HO-PCL115 (IB004-125-01)
[0188] 600 mg Bn-PCL115 was added to a 25 mL high pressure reactor, and fully dissolved in 6 mL methanol. 60 mg Pd/C was added and the solution was pressurized to 500 PSI. The temperature was raised to 50 C., and the reaction was stopped after 48 hours. The solution was filtered, and the filtrate was slowly added with 60 ml of methyl tert-butyl ether. White precipitate appeared. The solution was stirred for 10 min and then filtered to obtain 496 mg of white solid polymer with a yield of 82.8%. .sup.1H NMR (400 MHZ, CDCl.sub.3) 4.27-4.13 (m, 342H), 3.66 (s, 448H), 2.18-2.00 (m, 240H), 1.85-1.42 (m, 492H). Mw: 28447, Mn: 23116, PDI: 1.231.
2.3 Side Chain Coupling
2.3.1 PCL38-TEPr (IB005-018-01)
[0189] In a single-necked flask, polymer PCL38 (25.4 mg, 0.002059 mmol) and 5 mL of dichloromethane were added, and magnetically stirred to dissolve. Then, cysteamine hydrochloride (0.702 mg, 0.006177 mmol) and diisopropylethylamine (DIPEA, 26.45 mg, 0.2046 mmol) were added, and the mixture was stirred at room temperature for 10 minutes. Compound 2 (the structure of which is shown in the corresponding flow chart below; 13.89 mg, 0.0756 mmol) was added and stirred overnight. The reaction solution was transferred and subjected to rotatory evaporation to remove the solvent, and 10 mL of 50% ethanol was added. After 24 hours of dialysis, the concentrated solution was subjected to rotatory evaporation to obtain the 26.2 mg of product with a yield of 86.9%. .sup.1H NMR (400 MHZ, CDCl.sub.3) 4.15 (t, J=6.6 Hz, 76H), 3.64 (s, 448H), 3.33 (t, J=7.2 Hz, 38H), 3.09-2.59 (m, 292H), 2.09-1.35 (m, 158H), 1.25 (t, J=7.8 Hz, 114H), 0.96 (t, J=7.2 Hz, 114H).
##STR00043##
2.3.2 PCL56-TEPr (IB005-011-01)
[0190] The synthesis and purification of example 2.3.2 were performed according to the procedure in example 2.3.1 above (change from PCL38 to PCL56 required adjusted molar amount of monomer needed for PCL56 and the corresponding molar quantity of Compound 2, shown in below scheme, was also adjusted to match the quantity of side chains) to obtain 140.4 mg of the product with a yield of 95.8, shown in below scheme. .sup.1H NMR (400 MHZ, CDCl.sub.3) 4.14 (t, J=6.6 Hz, 112H), 3.64 (s, 448H), 3.33 (t, J=7.2 Hz, 56H), 3.09-2.59 (m, 436H), 2.09-1.35 (m, 230H), 1.25 (t, J=7.8 Hz, 168H), 0.96 (t, J=7.2 Hz, 168H).
##STR00044##
2.3.3 PCL70-TEPr (IB005-012-01)
[0191] The synthesis and purification of example 2.3.3 were performed according to the procedure in example 2.3.1 above (change from PCL38 to PCL70 required adjusted molar amount of monomer needed for PCL70 and the corresponding molar quantity of Compound 2, shown in below scheme, was also adjusted to match the quantity of side chains) to obtain 138.8 mg of the product with a yield of 93.3%. .sup.1H NMR (400 MHZ, CDCl.sub.3) 4.14 (t, J=6.6 Hz, 140H), 3.65 (s, 448H), 3.33 (t, J=7.2 Hz, 70H), 3.08-2.58 (m, 548H), 2.09-1.35 (m, 286H), 1.24 (t, J=7.2 Hz, 210H), 0.96 (t, J=7.2 Hz, 210H).
##STR00045##
2.3.4 PCL83-TEPr (IB005-019-01)
[0192] The synthesis and purification of example 2.3.4 were performed according to the procedure in example 2.3.1 above (change from PCL38 to PCL83 required adjusted molar amount of monomer needed for PCL83 and the corresponding molar quantity of Compound 2, shown in below scheme, was also adjusted to match the quantity of side chains) to obtain 100.7 mg of the product with a yield of 95.8%. .sup.1H NMR (400 MHZ, CDCl.sub.3) 4.16 (t, J=6.6 Hz, 166H), 3.65 (s, 448H), 3.34 (t, J=7.2 Hz, 83H), 3.09-2.59 (m, 652H), 2.10-1.36 (m, 338H), 1.26 (t, J=7.8 Hz, 249H), 0.96 (t, J=7.2 Hz, 249H).
##STR00046##
2.3.5 PCL100-TEPr (IB005-020-01)
[0193] The synthesis and purification of example 2.3.5 were performed according to the procedure in example 2.3.1 above (change from PCL38 to PCL100 required adjusted molar amount of monomer needed for PCL100 and the corresponding molar quantity of Compound 2, shown in below scheme, was also adjusted to match the quantity of side chains) to obtain 100 mg of the product with a yield of 96.0%. .sup.1H NMR (400 MHZ, CDCl.sub.3) 4.15 (q, J=6.6 Hz, 204H), 3.65 (s, 448H), 3.32 (m, J=7.6 Hz, 86H), 3.08-2.51 (m, 654H), 2.18-1.09 (m, 1091H), 0.95 (m, 254H).
##STR00047##
2.3.6 PCL115-TEPr (IB005-010-01)
[0194] The synthesis and purification of example 2.3.6 were performed according to the procedure in example 2.3.1 above (change from PCL38 to PCL115 required adjusted molar amount of monomer needed for PCL115 and the corresponding molar quantity of Compound 2, shown in below scheme, was also adjusted to match the quantity of side chains) to obtain 99.8 mg of the product with a yield of 96.5%, .sup.1H NMR (400 MHz, CDCl.sub.3) 4.12 (t, J=6.6 Hz, 230H), 3.64 (s, 448H), 3.33 (t, J=7.2 Hz, 115H), 3.09-2.59 (m, 908H), 2.08-1.34 (m, 466H), 1.25 (t, J=7.2 Hz, 345H), 0.96 (t, J=7.2 Hz, 345H).
##STR00048##
2.3.7 PCL150-TEPr (IB005-025-01)
[0195] The synthesis and purification of example 2.3.7 were performed according to the procedure in example 2.3.1 above (change from PCL38 to PCL150 required adjusted molar amount of monomer needed for PCL150 and the corresponding molar quantity of Compound 2, shown in below scheme, was also adjusted to match the quantity of side chains) to obtain 100.6 mg of the product with a yield of 97.6%. .sup.1H NMR (400 MHz, CDCl.sub.3) 4.12 (t, J=6.6 Hz, 314H), 3.65 (s, 448H), 3.33 (t, J=7.2 Hz, 157H), 3.09-2.59 (m, 1244H), 2.09-1.35 (m, 634H), 1.25 (t, J=7.8 Hz, 471H), 0.96 (t, J=7.2 Hz, 471H).
##STR00049##
2.3.8 PCL180-TEPr (IB005-025-01)
[0196] The synthesis and purification of example 2.3.8 were performed according to the procedure in example 2.3.1 above (change from PCL38 to PCL180 required adjusted molar amount of monomer needed for PCL180 and the corresponding molar quantity of Compound 2, shown in below scheme, was also adjusted to match the quantity of side chains) to obtain 101.5 mg of the product with a yield of 97.8%. .sup.1H NMR (400 MHz, CDCl.sub.3) 4.15 (t, J=6.6 Hz, 376H), 3.65 (s, 448H), 3.34 (t, J=7.6 Hz, 188H), 3.09-2.59 (m, 1492H), 2.09-1.35 (m, 758H), 1.25 (t, J=7.8 Hz, 564H), 0.96 (t, J=7.2 Hz, 564H).
##STR00050##
2.3.9 PCL115-TEE
[0197] The synthesis and purification of example 2.3.9 were performed according to the procedure in example 2.3.1 above (change from PCL38 to PCL115 required adjusted molar amount of monomer needed for PCL115 and the corresponding molar quantity of Compound 2, shown in below scheme, was also adjusted to match the quantity of side chains) to obtain 135 mg of the product with a yield of 92.3%. .sup.1H NMR (400 MHZ, CDCl.sub.3) 4.06 (t, J=6.6 Hz, 224H), 3.78 (s, 448H), 3.26 (t, J=7.2 Hz, 112H), 2.94-2.51 (m, 908H), 2.19-1.40 (m, 672H), 1.21 (t, J=7.8 Hz, 672H).
##STR00051##
2.3.10 PCL115-TEE43-TEPr69
[0198] The synthesis and purification of example 2.3.10 were performed according to the procedure in example 2.3.1 above (change from PCL38 to PCL115 required adjusted molar amount of monomer needed for PCL115, and the corresponding molar quantities of Compound 2 and Compound 3, shown in below scheme, were also adjusted respectively to match the quantities of desired side chains) to obtain 134 mg of the product with a yield of 90.6%. .sup.1H NMR (400 MHZ, CDCl.sub.3) 4.13 (t, J=6.6 Hz, 200H), 3.78 (s, 448H), 3.43 (t, J=7.2 Hz, 100H), 2.94-2.46 (m, 788H), 2.19-1.30 (m, 648H), 1.26 (t, J=7.8 Hz, 510H), 0.96 (t, J=7.2 Hz, 72H).
##STR00052##
2.3.11 PCL115-TEE62-TEPr50
[0199] The synthesis and purification of example 2.3.11 were performed according to the procedure in example 2.3.1 above (change from PCL38 to PCL115 required adjusted molar amount of monomer needed for PCL115, and the corresponding molar quantities of Compound 2 and Compound 3, shown in below scheme, were also adjusted respectively to match the quantities of desired side chains) to obtain 92.6 mg of the product with a yield of 91.8%. .sup.1H NMR (400 MHZ, CDCl.sub.3) 4.02 (t, J=6.6 Hz, 230H), 3.78 (s, 448H), 3.49 (t, J=7.2 Hz, 115H), 3.15-2.67 (m, 908H), 2.11-1.34 (m, 793H), 1.25 (t, J=7.8 Hz, 518H), 0.94 (t, J=7.2 Hz, 154H).
##STR00053##
2.3.12 PCL115-TPrPr
[0200] The synthesis and purification of example 2.3.12 were performed according to the procedure in example 2.3.1 above (change from PCL38 to PCL115 required adjusted molar amount of monomer needed for PCL115 and the corresponding molar quantity of Compound 2, shown in below scheme, was also adjusted to match the quantity of side chains) to obtain 122.5 mg of the product with a yield of 90.6%. .sup.1H NMR (400 MHz, CDCl.sub.3) 3.83 (t, J=6.6 Hz, 200H), 3.75 (s, 448H), 3.42 (t, J=7.2 Hz, 100H), 2.98-2.53 (m, 788H), 2.20-1.31 (m, 988H), 1.08 (t, J=7.2 Hz, 582H).
##STR00054##
2.3.13 HO-PCL115-TEPr
[0201] The synthesis and purification of example 2.3.13 were performed according to the procedure in example 2.3.1 above (change from PCL38 to PCL115 required adjusted molar amount of monomer needed for PCL115 and the corresponding molar quantity of Compound 2, shown in below scheme, was also adjusted to match the quantity of side chains) to obtain 103.7 mg of the product with a yield of 97.0%. .sup.1H NMR (400 MHz, CDCl.sub.3) 4.13 (t, J=6.6 Hz, 230H), 3.66 (s, 448H), 3.34 (t, J=7.2 Hz, 115H), 3.11-2.59 (m, 908H), 2.09-1.33 (m, 466H), 1.25 (t, J=7.2 Hz, 345H), 0.96 (t, J=7.2 Hz, 345H).
##STR00055##
2.3.14 PCL115-TEPr-ALA10
[0202] The synthesis and purification of example 2.3.14 were performed according to the procedure in example 2.3.1 above (change from PCL38 to PCL115 required adjusted molar amount of monomer needed for PCL115, and the corresponding molar quantities of Compound 2 and Compound 3, shown in below scheme, were also adjusted respectively to match the quantities of desired side chains) to obtain 120.8 mg of the product with a yield of 95.5%, .sup.1H NMR (400 MHZ, CDCl.sub.3) 4.13-4.01 (t, J=6.6 Hz, 250H), 3.66 (br, 468H), 3.33 (t, J=7.2 Hz, 115H), 3.11-2.59 (m, 856H), 2.09-1.33 (m, 455H), 1.25 (m, 342H), 0.96 (t, J=7.2 Hz, 306H).
##STR00056##
2.4 Coupling of Fluorescent Molecules
2.4.1 PCL38-TEPr-ICG3 (IB005-021-01)
[0203] In a single-necked flask, PCL38-TEPr (26.2 mg, 0.00179 mmol) and 3 mL DCM were added, and magnetically stirred to get dissolved. ICG-OSu (8.87 mg, 0.0107 mmol) and DIPEA (1.39 mg, 0.0107 mmol) solutions were added and stirred overnight at room temperature. The solvent was removed by rotary evaporation of the reaction solution and 10 mL of 50% ethanol (containing one drop of trifluoroacetic acid) was added. Ultrafiltration and centrifugal purification were repeated three times. The obtained concentrated liquid was rotary evaporated to dryness to obtain the product trifluoroacetate (33.9 mg, 0.00164 mmol) with a yield of 91.6%. .sup.1H NMR (400 MHz, CDCl.sub.3) 8.12-7.47 (m, 45H), 6.74-6.44 (m, 12H), 4.14 (t, J=6.4 Hz 76H), 3.66 (s, 448H), 3.34 (t, J=7.2 Hz, 38H), 3.42-3.17 (m, 292H), 2.09-1.35 (m, 242H), 1.25 (t, J=7.6 Hz, 114H), 1.02 (t, J=7.2 Hz, 114H).
##STR00057##
2.4.2 PCL56-TEPr-ICG3 (IB005-017-01)
[0204] The synthesis and purification of example 2.4.2 were performed according to the procedure in example 2.4.1 above (change from PCL38-TEPr to PCL115-TEPr required adjustment for proper molar ratio of polymer to ICG required per polymer chain) to obtain 67.2 mg of the product with a yield of 94.6%. .sup.1H NMR (400 MHZ, CDCl.sub.3) 8.12-7.47 (m, 45H), 6.72-6.44 (m, 12H), 4.13 (t, J=6.4 Hz 112H) 3.66 (s, 448H), 3.34 (t, J=7.2 Hz, 56H), 3.40-3.16 (m, 436H), 2.07-1.34 (m, 314H), 1.24 (t, J=7.6 Hz, 168H), 1.0 (t, J=7.2 Hz, 168H).
2.4.3 PCL70-TEPr-ICG3 (IB005-016-01)
[0205] The synthesis and purification of example 2.4.3 were performed according to the procedure in example 2.4.1 above (change from PCL38-TEPr to PCL70-TEPr required adjustment for proper molar ratio of polymer to ICG required per polymer chain) to obtain 138.8 mg of the product with a yield of 93.3%. .sup.1H NMR (400 MHZ, CDCl.sub.3) 8.14-7.48 (m, 45H), 6.74-6.44 (m, 12H), 4.15 (t, J=6.4 Hz 140H), 3.66 (s, 448H), 3.34 (t, J=7.2 Hz, 70H), 3.42-3.17 (m, 548H), 2.11-1.37 (m, 370H), 1.26 (t, J=7.2 Hz, 210H), 1.03 (t, J=7.2 Hz, 210H).
2.4.4 PCL83-TEPr-ICG3 (IB005-022-01)
[0206] The synthesis and purification of example 2.4.4 were performed according to the procedure in example 2.4.1 above (change from PCL38-TEPr to PCL83-TEPr required adjustment for proper molar ratio of polymer to ICG required per polymer chain) to obtain 100.7 mg of the product with a yield of 95.8%. .sup.1H NMR (400 MHZ, CDCl.sub.3) 8.11-7.45 (m, 45H), 6.75-6.44 (m, 12H), 4.13 (t, J=6.4 Hz 166H), 3.66 (s, 448H), 3.33 (t, J=7.2 Hz, 83H), 3.42-3.16 (m, 652H), 2.09-1.35 (m, 422H), 1.25 (t, J=7.2 Hz, 249H), 1.02 (t, J=7.2 Hz, 249H).
2.4.5 PCL100-TEPr-ICG3 (IB005-023-01)
[0207] The synthesis and purification of example 2.4.5 were performed according to the procedure in example 2.4.1 above (change from PCL38-TEPr to PCL100-TEPr required adjustment for proper molar ratio of polymer to ICG required per polymer chain) to obtain 100 mg of the product with a yield of 96.5%, .sup.1H NMR (400 MHZ, CDCl.sub.3) 8.14-7.45 (m, 45H), 6.76-6.45 (m, 12H), 4.15 (t, J=6.4 Hz 200H), 3.66 (s, 448H), 3.34 (t, J=7.2 Hz, 100H), 3.42-3.16 (m, 788H), 2.09-1.35 (m, 490H), 1.26 (t, J=7.2 Hz, 300H), 1.01 (t, J=7.2 Hz, 300H).
2.4.6 PCL115-TEPr-ICG3 (IB005-034-01)
[0208] The synthesis and purification of example 2.4.6 were performed according to the procedure in example 2.4.1 above (change from PCL38-TEPr to PCL115-TEPr required adjustment for proper molar ratio of polymer to ICG required per polymer chain) to obtain 60 mg of product with a yield of 87.0%. .sup.1H NMR (400 MHz, CDCl.sub.3) 8.12-7.46 (m, 45H), 6.74-6.43 (m, 12H), 4.14 (t, J=6.4 Hz 230H), 3.66 (s, 448H), 3.34 (t, J=7.2 Hz, 115H), 3.42-3.17 (m, 908H), 2.08-1.34 (m, 550H), 1.25 (t, J=7.6 Hz, 345H), 1.03 (t, J=7.2 Hz, 345H).
2.4.7 PCL150-TEPr-ICG3 (IB005-027-01)
[0209] The synthesis and purification of example 2.4.7 were performed according to the procedure in example 2.4.1 above (change from PCL38-TEPr to PCL150-TEPr required adjustment for proper molar ratio of polymer to ICG required per polymer chain) to obtain 67.00 mg of the product with a yield of 93.5%, .sup.1H NMR (400 MHZ, CDCl.sub.3) 8.12-7.45 (m, 45H), 6.74-6.44 (m, 12H), 4.14 (t, J=6.4 Hz 314H), 3.66 (s, 448H), 3.34 (t, J=7.2 Hz, 157H), 3.42-3.17 (m, 1244H), 2.09-1.35 (m, 718H), 1.25 (t, J=7.6 Hz, 471H), 1.02 (t, J=7.2 Hz, 471H).
2.4.8 PCL180-TEPr-ICG3 (IB005-026-01)
[0210] The synthesis and purification of example 2.4.8 were performed according to the procedure in example 2.4.1 above (change from PCL38-TEPr to PCL180-TEPr required adjustment for proper molar ratio of polymer to ICG required per polymer chain) to obtain 67.76 mg of the product with a yield of 94.6%. .sup.1H NMR (400 MHZ, CDCl.sub.3) 8.12-7.47 (m, 45H), 6.77-6.45 (m, 12H), 4.15 (t, J=6.4 Hz 376H), 3.66 (s, 448H), 3.35 (t, J=7.2 Hz, 188H), 3.42-3.18 (m, 1492H), 2.12-1.36 (m, 842H), 1.25 (t, J=7.2 Hz, 564H), 1.03 (t, J=7.2 Hz, 564H).
2.4.9 PCL115-TEE-ICG3 (IB004-090-01)
[0211] The synthesis and purification of example 2.4.9 were performed according to the procedure in example 2.4.1 above (change from PCL38-TEPr to PCL115-TEPr required adjustment for proper molar ratio of polymer to ICG required per polymer chain) to obtain 49.3 mg of the product with a yield of 91.7%. .sup.1H NMR (400 MHZ, CDCl.sub.3) 8.18-7.54 (m, 45H), 6.78-6.46 (m, 12H), 4.16 (t, J=6.6 Hz, 224H), 3.67 (s, 448H), 3.36 (t, J=7.2 Hz, 112H), 2.96-2.46 (m, 908H), 2.19-1.40 (m, 690H), 1.21 (t, J=7.8 Hz, 672H).
##STR00058##
2.4.10 PCL115-TEE62-TEPr50-ICG3
[0212] The synthesis and purification of example 2.4.10 were performed according to the procedure in example 2.4.1 above (change from PCL38-TEPr to PCL115-TEE62-TEPr50 required adjustment for proper molar ratio of polymer to ICG required per polymer chain) to obtain 44.8 mg of the product with a yield of 93.6%. .sup.1H NMR (400 MHZ, CDCl.sub.3) 8.13-7.54 (m, 45H), 6.75-6.45 (m, 12H), 4.13 (t, J=6.6 Hz, 200H), 3.78 (s, 448H), 3.43 (t, J=7.2 Hz, 100H), 2.94-2.46 (m, 908H), 2.19-1.30 (m, 690H), 1.26 (t, J=7.8 Hz, 510H), 0.96 (t, J=7.2 Hz, 150H).
##STR00059##
2.4.11 PCL115-TEE43-TEPr69-ICG3
[0213] The synthesis and purification of example 2.4.11 were performed according to the procedure in example 2.4.1 above (change from PCL38-TEPr to PCL115-TEE43-TEPr69 required adjustment for proper molar ratio of polymer to ICG required per polymer chain) to obtain 50.1 mg of the product with a yield of 93.5%. .sup.1H NMR (400 MHZ, CDCl.sub.3) 8.14-7.54 (m, 45H), 6.74-6.44 (m, 12H), 4.11 (t, J=6.6 Hz, 230H), 3.78 (s, 448H), 3.45 (t, J=7.2 Hz, 115H), 3.15-2.68 (m, 908H), 2.12-1.35 (m, 793H), 1.25 (t, J=7.8 Hz, 603H), 0.95 (t, J=7.2 Hz, 207H).
##STR00060##
2.4.12 PCL115-TPrPr-ICG3
[0214] The synthesis and purification of example 2.4.12 were performed according to the procedure in example 2.4.1 above (change from PCL38-TEPr to PCL115-TPrPr required adjustment for proper molar ratio of polymer to ICG required per polymer chain) to obtain 62.00 mg of the product with a yield of 90.7%. .sup.1H NMR (400 MHZ, CDCl.sub.3) 8.16-7.49 (m, 45H), 6.75-6.47 (m, 12H), 4.13 (t, J=6.6 Hz, 200H), 3.75 (s, 448H), 3.42 (t, J=7.2 Hz, 100H), 2.98-2.53 (m, 788H), 2.20-1.31 (m, 1078H), 0.95 (t, J=7.2 Hz, 672H).
##STR00061##
2.4.13 PCL115-TEPr-ICG0.5
[0215] The synthesis and purification of example 2.4.134 were performed according to the procedure in example 2.4.1 above (change from PCL38-TEPr to PCL115-TEPr required adjustment for proper molar ratio of polymer to ICG required per polymer chain, note that 0.5 ICG per polymer chain means that average ICG among polymer chains, i.e. some of the polymers are 0 and some of polymers are 1 and more) to obtain 65.8 mg of the dark green solid product with the yield of 94.5%, .sup.1H NMR (400 MHZ, CDCl.sub.3) 8.12-7.45 (m, 8H), 6.81-6.47 (m, 2H), 4.18 (t, J=6.4 Hz 230H), 3.49 (s, 448H), 3.16 (t, J=7.2 Hz, 115H), 3.09-2.49 (m, 918H), 2.19-1.39 (m, 934H), 1.25 (t, J=7.6 Hz, 344H), 0.94 (t, J=7.2 Hz, 336H). 2.4.14 PCL115-TEPr-ICG1
[0216] The synthesis and purification of example 2.4.14 were performed according to the procedure in example 2.4.1 above (change from PCL38-TEPr to PCL115-TEPr required adjustment for proper molar ratio of polymer to ICG required per polymer chain, and the molar amount of ICG-OSu was changed to 1.0 per polymer chain) to obtain the product 65.5 mg with the yield of 93.6%. .sup.1H NMR (400 MHZ, CDCl.sub.3) 8.12-7.45 (m, 15H), 6.74-6.48 (m, 4H), 4.14 (t, J=6.4 Hz 230H), 3.82 (s, 448H), 3.16 (t, J=7.2 Hz, 115H), 3.10-2.49 (m, 916H), 2.05-1.40 (m, 948H), 1.29 (t, J=7.6 Hz, 342H), 0.94 (t, J=7.2 Hz, 336H).
2.4.15 PCL115-TEPr-ICG2
[0217] The synthesis and purification of example 2.4.15 were performed according to the procedure in example 2.4.1 above (change from PCL38-TEPr to PCL115-TEPr required adjustment for proper molar ratio of polymer to ICG required per polymer chain, and the molar amount of ICG-OSu was changed to 2.0 per polymer chain) to obtain the product 64.78 mg, .sup.1H NMR (400 MHZ, CDCl.sub.3) 8.12-7.45 (m, 30H), 6.74-6.46 (m, 8H), 4.16 (t, J=6.4 Hz 230H), 3.75 (s, 448H), 3.20 (t, J=7.2 Hz, 115H), 3.09-2.54 (m, 912H), 2.15-1.28 (m, 976H), 1.19 (t, J=7.6 Hz, 339H), 0.94 (t, J=7.2 Hz, 339H).
2.4.16 PCL115-TEPr-ICG4
[0218] The synthesis and purification of example 2.4.16 were performed according to the procedure in example 2.4.1 above (change from PCL38-TEPr to PCL115-TEPr required adjustment for proper molar ratio of polymer to ICG required per polymer chain, and the molar amount of ICG-OSu was changed to 4.0 per polymer chain) to obtain the product 66.1 mg, .sup.1H NMR (400 MHZ, CDCl.sub.3) 8.18-7.44 (m, 60H), 6.74-6.38 (m, 16H), 4.17 (t, J=6.4 Hz 230H), 3.75 (s, 448H), 3.20 (t, J=7.2 Hz, 115H), 3.09-2.54 (m, 904H), 2.09-1.31 (m, 1032H), 1.19 (t, J=7.6 Hz, 333H), 0.93 (t, J=7.2 Hz, 333H).
2.4.17 PCL115-TEPr-ICG5
[0219] The synthesis and purification of example 2.4.17 were performed according to the procedure in example 2.4.1 above (change from PCL38-TEPr to PCL115-TEPr required adjustment for proper molar ratio of polymer to ICG required per polymer chain, and the molar amount of ICG-OSu was changed to 5.0 per polymer chain) to obtain the product 68.3 mg with the yield of 91.6%. .sup.1H NMR (400 MHZ, CDCl.sub.3), 8.14-7.50 (m, 75H), 6.81-6.49 (m, 20H), 4.16 (t, J=6.4 Hz 230H), 3.75 (s, 448H), 3.21 (t, J=7.2 Hz, 115H), 3.03-2.59 (m, 900H), 2.19-1.30 (m, 1060H), 1.19 (t, J=7.6 Hz, 330H), 0.95 (t, J=7.2 Hz, 330H).
2.4.18 HO-PCL115-TEPr-ICG3
[0220] The synthesis and purification of example 2.4.18 were performed according to the procedure in example 2.4.1 above (change from PCL38-TEPr to HO-PEG-PCL115-TEPr required adjustment for proper molar ratio of polymer to ICG required per polymer chain) to obtain 63 mg of the product with a yield of 89.6%. .sup.1H NMR (400 MHZ, CDCl.sub.3) 8.12-7.46 (m, 45H), 6.76-6.43 (m, 12H), 4.13 (t, J=6.4 Hz 230H), 3.66 (s, 448H), 3.33 (t, J=7.2 Hz, 115H), 3.43-3.17 (m, 908H), 2.09-1.34 (m, 550H), 1.25 (t, J=7.6 Hz, 345H), 0.97 (t, J=7.2 Hz, 345H).
2.4.19 PCL115-TEPr-ALA10-ICG3
[0221] The synthesis of PCL115-TEPr-ALA10-ICG3 is shown in the figure below. The polymer PCL115-TEPr-Fmoc-ALA10 (125 mg, 0.0033 mmol) is dissolved in 2 ml DMF. ICG-Osu (5.7 mg, 0.0067 mmol) and DIEA (25 mg, 0.2 mmol) were added. After the addition, stir overnight at room temperature, remove DIEA by rotary evaporation and add 0.2 ml piperidine. The solution was stirred at room temperature for 0.5 hours, DMF is concentrated, and the residue is dissolved in 100 ml absolute ethanol. A ceramic membrane (5K) is used to purify for 2 hours. The solution is concentrated to remove EtOH, and dried under vacuum to obtain 110 mg of polymer, which was dark green solid. The synthesis and purification of example 2.4.19 were performed according to the process of example 2.4.1 above (PCL38 was replaced with an equimolar amount of PCL115-TEPr-ALA10) to obtain 100.4 mg of the product with the yield of 89.6%. .sup.1H NMR (400 MHZ, CDCl.sub.3) 8.12-7.45 (m, 45H), 6.75-6.42 (m, 12H), 4.13-4.01 (t, J=6.6 Hz, 250H), 3.66 (br, 468H), 3.34 (t, J=7.2 Hz, 115H), 3.10-2.57 (m, 856H), 2.09-1.34 (m, 455H), 1.25 (m, 342H), 0.96 (t, J=7.2 Hz, 306H).
##STR00062##
Example 2
pKa Test:
[0222] Accurately weigh 30 mg of the polymer prepared in example 1 (2.3.5, 2.3.7, 2.3.10), and dissolve it in 30 mL of 0.01 mol/L trifluoroacetic acid solution, and use 0.1 mol/L of sodium hydroxide to titrate under the instructions of the pH meter, and the volume of the sodium hydroxide solution consumed and the corresponding pH value are recorded. The volume is plotted against the pH value by Origin software. The pKa value is one-half of the sum of the two intersection points of the two tangent lines and the platform tangent. The specific results are shown in
Example 3
CMC Test of PCL Nano Fluorescent Probe:
[0223] Add 2 L of 110.sup.5 mol/L pyrene in tetrahydrofuran solution to PBS 8.0 solutions of the polymer at a series of concentrations (110.sup.6110.sup.1 mg/mL) (examples 2.3.5 and 2.3.10)), use a vortex mixer to mix evenly, and let it stabilize to test the fluorescence intensity of the solution. By plotting the fluorescence intensity ratio (1590/1660) against the concentration, the critical micelle concentration is determined as the intersection of two tangent lines. The CMC test results of two representative polymers are shown in
Example 4
4.1 Preparation and Characterization of Nanoparticle Solution:
[0224] 5 mg polymer was dissolved in 0.2 ml CH.sub.3CN, added to 5 ml deionized water under ultrasonic conditions, CH.sub.3CN was concentrated on a rotary evaporator, and deionized water was added to a volume of 5 ml, and the concentration of the obtained stock solution was 1 mg/ml.
4.2 DLS Test:
[0225] The sample used in this example is the same as that in example 2. PCL100-TEPr was used to prepare the nanoparticle solution. The pH of the solution was about 8.0, the concentration was 1 mg/mL, and the sample was taken at room temperature (20 C.) for DLS (The instrument is: Brookhaven Omni Dynamic Light Scattering (DLS) Particle Sizer and zeta potential Analyzer, all other DLS tests are measured on Malvern Zetasizer Ultra, HeNe laser, 2=633 nm). The data obtained is shown in
[0226] The nanoparticle solution of example 4.1 was dropped into PBS 6.0, the sample was shaken for 2 minutes and then the DLS test was performed. The data obtained is shown in
4.3 TEM Test:
[0227] Use PCL100-TEPr-ICG to prepare the nanoparticle solution. The concentration of this solution is 1 mg/mL, the pH is about 8.0, and the sample is taken for TEM test (ThermoFisher Scientific (formerly FEI), model: Talos F200S, origin: Netherlands). The data is shown in
[0228] The above-mentioned nanoparticle solution was dropped into the PBS6.0 solution for TEM test, and the obtained data is shown in
Example 5
5.1 Fluorescence Test:
[0229] 100 L nanoparticle stock solution (1 mg/mL, refer to example 4.1 for preparation method) was diluted into 2.0 mL of PBS buffer (pH 5.5-8.0), mixed well and emission fluorescence was measured. The excitation light wavelength is 730 nm, and the emission light wavelength detection range is 785-900 nm. The properties of PCL series fluorescent probes are shown in Table 1, where:
[0230] pKa and CMC measurement methods refer to Examples 2 and 3.
[0231] The calculation of the fluorescence intensity ratio (FIR) is the ratio of the fluorescence intensity at 821 nm in the pH 6.0 buffer solution of the nano fluorescent probe to the fluorescence intensity at 821 nm in the pH 8.0 buffer solution. The calculation method is as follows:
[0232] Calculation of pH transition point (pHt): Take the fluorescence intensity at 821 nm of different pH values, perform mathematical normalization, and plot the pH versus fluorescence intensity. Then the obtained scatter plot is fitted with boltzmann function. The pH value at 50% fluorescence intensity of the highest fluorescence value is pHt. [0233] pH.sub.50%. The calculation method of pH mutation range is as follows:
TABLE-US-00001 TABLE 1 Screening of nano fluorescent PCL probes Example Tertiary amine ICG/ pH number DP side chain Chain FIR pHt 10-90% 2.4.9 115 TEE 3 22 7.19 0.32 IB005-015-06 115 TEPr 3 129 6.59 0.03 2.4.10 115 TEE62-TEPr50 3 31 7.01 0.15 2.4.11 115 TEE43-TEPr69 3 45 6.73 0.43 2.4.12 115 TPrPr 3 49 5.69 0.19 2.4.1 38 TEPr 3 46 6.51 0.46 2.4.2 56 TEPr 3 50 6.60 0.30 2.4.3 70 TEPr 3 96 6.61 0.37 2.4.4 83 TEPr 3 48 6.52 0.20 2.4.5 100 TEPr 3 42 6.37 0.29 2.4.7 150 TEPr 3 44 6.39 0.34 2.4.8 180 TEPr 3 64 6.37 0.33 2.4.13 115 TEPr 0.5 4.5 6.37 0.34 2.4.14 115 TEPr 1 20 6.44 0.27 2.4.15 115 TEPr 2 41 6.43 0.31 2.4.16 115 TEPr 4 70 6.47 0.42 2.4.17 115 TEPr 5 67 6.42 0.36
5.2 the Effect of the Degree of Polymerization of the Hydrophobic Block (PCL) on FIR, pHt and pH
[0234] The fluorescence test of the nanoparticle stock solution (1 mg/mL, the preparation method refers to example 4.1) was implemented with reference to example 5.1. The relationship between the fluorescence emission intensity of PCL-TEPr-ICG3 with different degree of polymerization (DP) and pH at 821 nm is summarized in Table 1 and
5.3 the Effect of Side Chain Tertiary Amine on FIR, pHt and pH of PCL100-ICG3
[0235] The fluorescence test of the nanoparticle stock solution (1 mg/mL, the preparation method refers to example 4.1) was implemented with reference to example 5.1. The relationship between the fluorescence emission intensity of the PCL100-ICG3 probe with TEE, TEPr, TPRPr, TEE62-TEPr50 or TEE43-TEPr69 connected to the side chain at 821 nm and pH is summarized in Table 1 and
5.4 the Effect of the Number of ICG on the FIR, pHt and pH of PCL100-TEPr
[0236] Summary of the relationship between the fluorescence emission intensity of the PCL100-TEPr probes (with different numbers of ICG attached to the side chain, and 0.5, 1.0, 2.0, 3.0, 4.0, 5.0 ICG attached to each polymer side chain) at 821 nm and pH are shown in Table 1 and
Example 6
6.1 In Vivo Imaging Experiment of Subcutaneous Colorectal Cancer Tumor in Mice:
[0237] The colorectal cancer tumor was implanted on the lower right side of the back of the NPG mouse. When the tumor volume grew to 300-400m.sub.3, the mouse was anesthetized, and the PCL115-TEPr-ICG3 nanoparticle solution (2.5 mg/mL) was injected into the mouse through the tail vein. At different time points, a real-time fluorescence imaging system (PerkinElmer, model: IVIS spectrum CT, place of production: USA) was used to monitor the distribution of fluorescent probes in the body. As shown in
6.2 In Vivo Imaging Experiment of In Situ Bladder Cancer Tumor in Rats:
[0238] For in situ tumors in SD rats (the tumor formation method is chemical toxin perfusion induction; the tumors formed occupy more than 50% of the rat bladder volume), after the rats are anesthetized, 1 mg/mL PCL115-TEPr-ICG3 nanoparticle solution (2 mg/mL) was injected into the mouse's bladder via a catheter. At different time points, a real-time fluorescence imaging system (PerkinElmer, model: IVIS spectrum CT, origin: the United States) was used to monitor the distribution of fluorescent probes in the body. Immediately after perfusion, imaging showed that there was no fluorescence in the bladder. Two hours after perfusion, fluorescent imaging signals appeared in the bladder. About 5 hours after perfusion, the rats were sacrificed. After that, the rats were dissected on site, and their bladders were removed and measured. After performing the front and back fluorescence imaging, the bladder was further stripped and layered to obtain the serosal layer, muscle layer, and tumor, on which fluorescence imaging was performed, and further fluorescence quantitative analysis of each tissue was also achieved. The specific results are shown in
S1 General Preparation Procedure of Hydrophilic and Hydrophobic Side Chains (E2 Group)
##STR00063##
[0239] As depicted in above reaction scheme, reactant polymer is fully dissolved in suitable solvents (including but not being limited to water, ethanol, acetonitrile, DMF, DMSO etc.), followed by the addition of E2 group reactants along with various amount of bases (including but not being limited to Et3N, DIPEA, Potassium Carbonate, Sodium Ethoxide, Cesium Carbonate etc.). The reaction solution is stirred at common laboratory room temperature for 3-5 hours. The solution is filtered, and precipitated out in MTBE to obtain crude product, which then is dried under vacuum to obtain white solid polymer.
S2 General Preparation Procedure of Drug Delivery Group (D2 Group)
##STR00064##
[0240] As depicted in above reaction scheme, reactant polymer is fully dissolved in suitable solvents (including but not being limited to water, ethanol, acetonitrile, DMF, DMSO etc.), followed by the addition of D2 group reactants along with various amount of bases (including but not being limited to Et3N, DIPEA, Potassium Carbonate, Sodium Ethoxide, Cesium Carbonate etc.). The reaction solution is stirred at common laboratory room temperature for 3-5 hours. The solution is filtered, and precipitated out in MTBE to obtain crude product, which then is dried under vacuum to obtain white solid polymer.
S3 General Preparation Procedure of Cholesterol and Its' Derivatives, Vitamin D, and Vitamin E Side Chains
##STR00065## ##STR00066## ##STR00067##
[0241] n is an integer ranging from 2 to 18.
[0242] As depicted in above reaction scheme, in a single flask bottle, Cholesterol and Its' derivatives, Vitamin D, and Vitamin E, are functionalized by reacting the HO-group to a Mercapto-alkanoic (C2-C18)-carboxylic acid in Toluene heated under Nitrogen for 12 hours. The reaction solution is then filtered and dried under vacuum to remove solvent. Final pure product was purified by column chromatography for future use to conjugate to polymer as side chains.
Example 7
7.1 PCL103-C9 (C.sub.9H.sub.19)-TPrPr-ICG
7.1.1 Synthetic Route of PCL103-C9-TPrPr-ICG
##STR00068##
7.1.2 Synthesis of PCL103 (IB008-092)
[0243] The synthesis and purification of example 7.1.2 were performed according to the procedure in example 2.2.1 above (using 396 mg 3-bromocaprolactone, 2.06 mmol) to obtain a total of 449 mg white solid polymer with a yield of 90%. .sup.1H NMR (400 MHZ, Chloroform-d) 4.22-4.09 (m, 308H), 3.62 (s, 448H), 2.15-1.93 (m, 226H), 1.78-1.37 (m, 425H). Mw: 24655 Mn: 19093 PDI: 1.29.
7.1.3 Synthesis of PCL103-C9-TPrPr (IB008-097)
[0244] The synthesis and purification of Example 7.1.3 was performed according to the procedure as in Example 2.3.1 (change from PCL38 to PCL103 required adjusted molar amount of monomer needed for PCL103 and the corresponding molar quantity of Compound 2) and Example S1 (using potassium carbonate with DMF) to obtain 86.1 mg of product with a yield of 95.0%. .sup.1H NMR (400 MHZ, Chloroform-d) 4.23-4.04 (m, 202H), 3.64 (s, 448H), 3.30-3.15 (m, 93H), 2.74-2.61 (m, 334H), 2.47-2.34 (m, 399H), 1.87 (dd, J=13.8, 7.7 Hz, 101H), 1.78-1.34 (m, 995H), 1.26 (d, J=3.8 Hz, 188H), 0.88 (t, J=7.4 Hz, 611H).
7.1.4 Synthesis of PCL103-C9-TPrPr-ICG (IB008-103)
[0245] Synthesis and purification of Example 7.1.4 was performed according to the procedure as in Example 2.4.1 above (change from PCL38-TEPr to PCL103-C9-TPrPr required adjusted molar amount of monomer needed for PCL103-C9-TPrPr) to obtain 86.1 mg of product with a yield of 95.0%. .sup.1H NMR (400 MHZ, Chloroform-d) 7.91-7.39 (m, 18H), 4.23-4.04 (m, 202H), 3.64 (s, 448H), 3.30-3.15 (m, 93H), 2.74-2.61 (m, 334H), 2.47-2.34 (m, 399H), 1.87 (dd, J=13.8, 7.7 Hz, 101H), 1.78-1.34 (m, 995H), 1.26 (d, J=3.8 Hz, 188H), 0.88 (t, J=7.4 Hz, 611H).
7.2 PCL110-C9-TPrPr-ICG
7.2.1 Synthetic Route of PCL110-C9-TPrPr-ICG
##STR00069##
7.2.2 Synthesis of PCL110 (IB008-165)
[0246] The synthesis and purification of example 7.2.2 PCL110 were performed according to the procedure in example 2.2.1 above (using 422 mg 3-bromocaprolactone, 2.20 mmol) to obtain a total of 478 mg white solid polymer with a yield of 89%. .sup.1H NMR (400 MHZ, Chloroform-d) 4.26-4.10 (m, 331H), 3.63 (s, 448H), 2.16-1.95 (m, 253H), 1.80-1.38 (m, 482H).
7.2.3 Synthesis of PCL110-C9 (IB008-167)
[0247] The synthesis and purification of example 7.1.3 PCL110-C9 were performed according to the procedure in example S1 above (using potassium carbonate with DMF). PCL110 (600 mg, 0.0234 mmol) yielded product (650 mg, 0.02 mmol) with a yield of 86.7% .sup.1H NMR (400 MHZ, Chloroform-d) 4.26-4.05 (m, 284H), 3.63 (d, J=1.8 Hz, 448H), 3.19 (t, J=7.6 Hz, 39H), 2.57 (q, J=7.3 Hz, 78H), 2.05 (tt, J=14.9, 9.1, 8.6 Hz, 146H), 1.88 (s, 68H), 1.78-1.16 (m, 1141H), 0.91-0.83 (m, 120H).
7.2.4 Synthesis of PCL110-C9-TPrPr (IB008-176)
[0248] The synthesis and purification of example 7.2.4 PCL110-C9-TPrPr were performed according to the procedure in example 2.3.1 above. PCL110-C9 (50 mg, 0.0017 mmol) yielded product (35 mg, 0.00071 mmol) with a yield of 42%. .sup.1H NMR (400 MHZ, Chloroform-d) 4.11 (q, J=6.1, 5.7 Hz, 209H), 3.64 (d, J=1.9 Hz, 448H), 3.21 (dt, J=18.8, 7.4 Hz, 97H), 2.75-2.50 (m, 315H), 2.38 (dd, J=8.6, 6.4 Hz, 220H), 2.09-1.81 (m, 290H), 1.76-1.14 (m, 1764H), 0.87 (dd, J=8.4, 6.4 Hz, 584H).
7.2.5 Synthesis of PCL110-C9-TPrPr-ICG
[0249] The synthesis and purification of example 7.2.5 PCL110-C9-TPrPr-ICG were performed according to the procedure in example 2.4.1 above to obtain product (32.1 mg) with a yield of 91.7%. .sup.1H NMR (400 MHZ, Chloroform-d) 7.67 (d, J=183.6 Hz, 20H), 4.11 (q, J=6.1, 5.7 Hz, 209H), 3.64 (d, J=1.9 Hz, 448H), 3.21 (dt, J=18.8, 7.4 Hz, 97H), 2.75-2.50 (m, 315H), 2.38 (dd, J=8.6, 6.4 Hz, 220H), 2.09-1.81 (m, 290H), 1.76-1.14 (m, 1764H), 0.87 (dd, J=8.4, 6.4 Hz, 584H).
7.3 PCL107-C9-TEE-ICG
7.3.1 Synthesis Route of PCL107-C9-TEE-ICG
##STR00070##
7.3.2 Synthesis of PCL107 (IB008-099)
[0250] Example 7.3.2 PCL107 was synthesized and purified according to the procedure as in Example 2.2.1 above. A total of 465 mg of white solid polymer was obtained in 92% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 4.29-4.09 (m, 321H), 3.64 (s, 448H), 2.18-1.96 (m, 241H), 1.82-1.39 (m, 460H).
7.3.3 Synthesis of PCL107-C9 (IB008-151)
[0251] The synthesis and purification of Example 7.3.3 PCL107-C9 were performed according to the procedure in Example S1 above (using potassium carbonate and DMF). A total of 148 mg of white solid polymer was obtained in 95% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 4.12 (p, J=7.4, 6.0 Hz, 212H), 3.63 (s, 448H), 3.18 (t, J=7.5 Hz, 81H), 2.57 (q, J=7.3 Hz, 186H), 1.97-1.80 (m, 139H), 1.68 (q, J=7.5 Hz, 334H), 1.61-1.47 (m, J=6.6 Hz, 291H), 1.25 (s, 1312H), 0.87 (t, J=6.7 Hz, 218H).
7.3.4 Synthesis of PCL107-C9-TEE (IB008-153)
[0252] The synthesis and purification of Example 7.3.4 PCL107-C9-TEE were performed according to the procedure in Example 2.3.1 to obtain a total of about 20 mg polymer. .sup.1H NMR (400 MHZ, Chloroform-d) 4.12 (p, J=7.4, 6.0 Hz, 212H), 3.63 (s, 448H), 3.18 (t, J=7.5 Hz, 81H), 3.04-2.72 (m, 221H), 2.57 (q, J=7.3 Hz, 186H), 1.97-1.80 (m, 139H), 1.68 (q, J=7.5 Hz, 334H), 1.61-1.47 (m, J=6.6 Hz, 291H), 1.25 (s, 1312H), 0.87 (t, J=6.7 Hz, 278H).
7.3.5 Synthesis of PCL107-C9-TEE-ICG
[0253] Example 7.3.5 PCL107-C9-TEE-ICG was synthesized and purified according to the procedure as in Example 2.4.1 above. A total of 25 mg of polymer was obtained in 85% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 8.09-7.34 (m, 19H), 4.12 (p, J=7.4, 6.0 Hz, 212H), 3.63 (s, 448H), 3.18 (t, J=7.5 Hz, 81H), 3.04-2.72 (m, 221H), 2.57 (q, J=7.3 Hz, 186H), 1.97-1.80 (m, 139H), 1.68 (q, J=7.5 Hz, 334H), 1.61-1.47 (m, J=6.6 Hz, 291H), 1.25 (s, 1312H), 0.87 (t, J=6.7 Hz, 278H).
Example 8
8. Example Series Containing C18 (C.sub.18H.sub.37):
8.1 PCL107-C18-TPrPr-ICG
8.1.1 Synthesis Route of PCL107-C18-TPrPr-ICG
##STR00071##
8.1.2 Synthesis of PCL107 (IB008-099)
[0254] Example 8.1.2 PCL107 was synthesized and purified according to the procedure as in Example 2.2.1 above. A total of 465 mg of white solid polymer was obtained in 92% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 4.29-4.09 (m, 321H), 3.64 (s, 448H), 2.18-1.96 (m, 241H), 1.82-1.39 (m, 460H).
8.1.3 Synthesis of PCL107-C18 (IB008-113)
[0255] Example 8.1.3 PCL107-C18 was synthesized and purified according to the procedure as in Example S1 above (using potassium carbonate and DMF). A total of 221 mg of white polymer was obtained in 94.6% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 4.26-4.05 (m, 296H), 3.63 (d, J=1.8 Hz, 448H), 3.19 (t, J=7.6 Hz, 25H), 2.57 (q, J=7.3 Hz, 50H), 2.05 (tt, J=14.9, 9.1, 8.6 Hz, 146H), 1.88 (s, 68H), 1.78-1.16 (m, 1381H), 0.91-0.83 (m, 75H).
8.1.4 Synthesis of PCL107-C18-TPrPr (IB008-139)
[0256] Synthesis and purification of Example 8.1.4 PCL107-C18-TPr were performed according to the procedure as in Example 2.3.1 above. A total of 24 mg of white polymer was obtained in 85% yield. .sup.1H NMR (400 MHz, Chloroform-d) 4.23-3.99 (m, 209H), 3.62 (s, 448H), 3.32 (d, J=8.0 Hz, 86H), 3.23-2.65 (m, 740H), 2.56 (q, J=7.3 Hz, 34H), 1.88 (dq, J=13.9, 7.0 Hz, 103H), 1.70 (dd, J=17.4, 10.3 Hz, 658H), 1.23 (s, 432H), 0.97 (t, J=6.9 Hz, 524H), 0.86 (t, J=6.6 Hz, 75H).
8.1.5 Synthesis of PCL107-C18-TPrPr-ICG
[0257] Synthesis and purification of Example 8.1.5 PCL107-C18-TPr-ICG were performed according to the procedure as in Example 2.4.1 above. A total of 21 mg of green polymer was obtained in 90% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 8.06-7.36 (m, 18H), 4.23-3.99 (m, 209H), 3.62 (s, 448H), 3.32 (d, J=8.0 Hz, 86H), 3.23-2.65 (m, 740H), 2.56 (q, J=7.3 Hz, 34H), 1.88 (dq, J=13.9, 7.0 Hz, 103H), 1.70 (dd, J=17.4, 10.3 Hz, 658H), 1.23 (s, 432H), 0.97 (t, J=6.9 Hz, 524H), 0.86 (t, J=6.6 Hz, 75H).
8.2 PCL107-C18-TEE-ICG
8.2.1 Synthesis Route of PCL107-C18-TEE-ICG
##STR00072##
8.2.2 Synthesis of PCL107 (IB008-099)
[0258] Example 8.2.2 PCL107 was synthesized and purified according to the procedure as in Example 2.2.1 above. A total of 465 mg of white solid polymer was obtained in 92% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 4.29-4.09 (m, 321H), 3.64 (s, 448H), 2.18-1.96 (m, 241H), 1.82-1.39 (m, 460H).
8.2.3 Synthesis of PCL107-C18 (IB008-150)
[0259] Example 8.2.3 PCL107-C18 was synthesized and purified according to the procedure as in Example S1 above (using potassium carbonate and DMF). A total of 199 mg of white polymer was obtained in 92% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 4.26-4.05 (m, 296H), 3.63 (d, J=1.8 Hz, 448H), 3.19 (t, J=7.6 Hz, 25H), 2.57 (q, J=7.3 Hz, 50H), 2.05 (tt, J=14.9, 9.1, 8.6 Hz, 146H), 1.88 (s, 68H), 1.78-1.16 (m, 1381H), 0.91-0.83 (m, 75H).
8.2.4 Synthesis of PCL107-C18-TEE (IB008-152)
[0260] Synthesis and purification of Example 8.2.4 PCL107-C18-TEE were performed according to the procedure as in Example 2.3.1 above. A total of 28 mg of white polymer was obtained in 84% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 4.11 (t, J=6.6 Hz, 230H), 3.63 (s, 488H), 3.21 (dt, J=26.5, 7.6 Hz, 99H), 2.79-2.60 (m, 261H), 2.54 (dq, J=14.5, 7.2 Hz, 317H), 1.88 (q, J=7.5 Hz, 117H), 1.68 (q, J=7.3 Hz, 342H), 1.53 (dp, J=13.2, 6.5 Hz, 231H), 1.24 (s, 1516H), 1.01 (t, J=7.2 Hz, 349H), 0.87 (t, J=6.7 Hz, 163H).
8.2.5 Synthesis of PCL107-C18-TEE-ICG (IB008-152)
[0261] Synthesis and purification of 8.2.5 PCL107-C18-TEE-ICG were performed according to the procedure as in Example 2.4.1 above. A total of 25 mg of green polymer was obtained in 83% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 8.16-7.32 (m, 40H), 4.12 (t, J=6.9 Hz, 211H), 3.64 (d, J=1.6 Hz, 448H), 3.22 (dt, J=28.3, 7.6 Hz, 96H), 2.89-2.49 (m, 544H), 2.35 (s, 139H), 1.89 (dq, J=15.0, 8.7, 7.3 Hz, 142H), 1.69 (h, J=6.9 Hz, 321H), 1.53 (dq, J=12.6, 6.8, 6.3 Hz, 219H), 1.45-1.33 (m, 196H), 1.25 (s, 1287H), 1.06 (d, J=7.2 Hz, 323H), 0.87 (t, J=6.6 Hz, 158H).
Example 9
9. Example Series Containing Cholesterol (-CHOL):
[0262] The cholesterol-containing embodiments all require modification of the HO-groups of the cholesterol and hydrophobic vitamin molecules. Cholesterol side chains can be conjugated to polymer by converting HO-group of cholesterol to a Thiol-group, via reaction scheme below.
Thioethyl Cholesterol
Synthetic Route of Thioethyl Cholesterol
##STR00073##
Synthesis of Thioethyl Cholesterol (IB008-114)
[0263] In this example, the synthesis and purification of thioethyl cholesterol was carried out according to the procedure in Example S3 above to obtain a total of 3 mg of white solid in 90% yield.
[0264] .sup.1H NMR (400 MHZ, Chloroform-d) 5.20-5.12 (m, 1H), 4.40 (d, J=9.8 Hz, 1H), 3.02 (d, J=8.2 Hz, 2H), 2.56 (s, 3H), 2.12 (d, J=8.3 Hz, 2H), 1.87-1.54 (m, 7H), 1.48-0.53 (m, 39H), 0.46 (s, 4H).
9.1 PCL107-CHOL-TPrPr-ICG
9.1.1 Synthesis Route of PCL107-CHOL-TPrPr-ICG
##STR00074##
9.1.2 Synthesis of PCL107 (IB008-099)
[0265] Example 9.1.2 PCL107 was synthesized and purified according to the procedure as in Example 2.2.1 above. A total of 465 mg of white solid polymer was obtained in 92% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 4.29-4.09 (m, 321H), 3.64 (s, 448H), 2.18-1.96 (m, 241H), 1.82-1.39 (m, 460H).
9.1.3 Synthesis of PCL107-CHOL (IB008-137)
[0266] Example 9.1.3 PCL107-CHOL was synthesized and purified according to the procedure as in Example S1 above. (using DIEA and dichloromethane), a total of 221 mg of white polymer was obtained in 94.6% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 5.38 (s, 5H), 4.34-4.10 (m, 319H), 3.64 (d, J=1.7 Hz, 448H), 3.28 (d, J=14.3 Hz, 5H), 2.31 (q, J=8.1, 7.5 Hz, 14H), 2.17-1.23 (m, 859H), 1.12 (s, 21H), 1.05-0.82 (m, 65H), 0.67 (s, 8H).
9.1.4 Synthesis of PCL107-CHOL-TPrPr (IB008-145)
[0267] Example 9.1.4 PCL107-CHOL-TPrPr was synthesized and purified according to the procedure as in Example 2.3.1 above. (using DIEA and dichloromethane), a total of 15 mg of white polymer was obtained in 92.6% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 5.38 (s, 5H), 4.27-4.00 (m, 230H), 3.62 (s, 448H), 3.23 (t, J=7.5 Hz, 92H), 2.71 (q, J=5.8 Hz, 351H), 2.55-2.38 (m, 362H), 1.93-1.79 (m, 114H), 1.75-1.61 (m, 315H), 1.58-1.33 (m, 702H), 0.87 (t, J=7.4 Hz, 586H).
9.1.5 Synthesis of PCL107-CHOL-TPrPr-ICG
[0268] Example 9.1.5 PCL107-CHOL-TPrPr-ICG was synthesized and purified according to the procedure as in Example 2.4.1 above. A total of 12 mg of green polymer was obtained in 80% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 8.06-7.30 (m, 20H), 5.38 (s, 5H), 4.27-4.00 (m, 230H), 3.62 (s, 448H), 3.23 (t, J=7.5 Hz, 92H), 2.71 (q, J=5.8 Hz, 351H), 2.55-2.38 (m, 362H), 1.93-1.79 (m, 114H), 1.75-1.61 (m, 315H), 1.58-1.33 (m, 702H), 0.87 (t, J=7.4 Hz, 586H).
9.2 PCL120-CHOL-TEPr-ICG
9.2.1 Synthesis Route of PCL120-CHOL-TEPr-ICG
##STR00075##
9.2.2 Synthesis of PCL120 (IB005-195)
[0269] Example 9.2.2 PCL120 was synthesized and purified according to the procedure as in Example 2.2.1 above. A total of 485 mg of white solid polymer was obtained in 96% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 4.26-4.11 (m, 360H), 3.64 (d, J=1.2 Hz, 448H), 2.06 (dddd, J=24.2, 14.7, 9.6, 6.3 Hz, 254H), 1.81-1.37 (m, 515H).
9.2.3 Synthesis of PCL120-CHOL (IB005-021)
[0270] Example 9.2.3 PCL120-CHOL was synthesized and purified according to the procedure as in Example S1 above. (using DIEA and dichloromethane), a total of 221 mg of white polymer was obtained in 94.6% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 5.38 (s, 10H), 4.34-4.10 (m, 319H), 3.64 (d, J=1.7 Hz, 448H), 3.28 (d, J=14.3 Hz, 10H), 2.31 (q, J=8.1, 7.5 Hz, 27H), 2.17-1.23 (m, 937H), 1.12 (s, 42H), 1.05-0.82 (m, 130H), 0.67 (s, 25H).
9.2.4 Synthesis of PCL120-CHOL-TEPr (IB005-029)
[0271] Example 9.2.4 PCL120-CHOL-TEPr was synthesized and purified according to the procedure as in Example 2.3.1 above. .sup.1H NMR (400 MHZ, Chloroform-d) 5.36, (s, 10H), 4.15 (dq, J=20.5, 8.4, 6.9 Hz, 238H), 3.63 (d, J=1.3 Hz, 448H), 3.33-3.22 (m, 99H), 2.91-2.46 (m, 810H), 2.12-1.81 (m, 163H), 1.76-1.35 (m, 983H), 1.13 (p, J=6.9 Hz, 349H), 0.95-0.82 (m, 382H), 0.66 (s, 28H).
9.2.5 Synthesis of PCL120-CHOL-TEPr-ICG
[0272] Example 9.2.5 PCL107-CHOL-TEPr-ICG was synthesized and purified according to the procedure as in Example 2.4.1 above. A total of 12 mg of green polymer was obtained in 80% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 8.06-7.30 (m, 19H), 5.36, (s, 10H), 4.15 (dq, J=20.5, 8.4, 6.9 Hz, 238H), 3.63 (d, J=1.3 Hz, 448H), 3.33-3.22 (m, 99H), 2.91-2.46 (m, 810H), 2.12-1.81 (m, 163H), 1.76-1.35 (m, 983H), 1.13 (p, J=6.9 Hz, 349H), 0.95-0.82 (m, 382H), 0.66 (s, 28H).
9.3 PCL120-CHOL-TPrPr-ICG
9.3.1 Synthesis Route of PCL120-CHOL-TPrPr-ICG
##STR00076##
9.3.2 Synthesis of PCL120 (IB005-195)
[0273] Example 9.3.2 PCL120 was synthesized and purified according to the procedure as in Example 2.2.1 above. A total of 485 mg of white solid polymer was obtained in 96% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 4.26-4.11 (m, 360H), 3.64 (d, J=1.2 Hz, 448H), 2.06 (dddd, J=24.2, 14.7, 9.6, 6.3 Hz, 254H), 1.81-1.37 (m, 515H).
9.3.3 PCL120-CHOL (IB005-019)
[0274] Example 9.3.3 PCL120-CHOL was synthesized and purified according to the procedure as in Example S1 above. (using DIEA and dichloromethane), a total of 52 mg of white polymer was obtained in 98% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 5.38 (s, 3H), 4.34-4.10 (m, 321H), 3.64 (d, J=1.7 Hz, 448H), 3.28 (d, J=14.3 Hz, 3H), 2.31 (q, J=8.1, 7.5 Hz, 11H), 2.17-1.23 (m, 859H), 1.12 (s, 21H), 1.05-0.82 (m, 65H), 0.67 (s, 6H).
9.3.4 Synthesis of PCL120-CHOL-TPrPr (IB005-027)
[0275] Example 9.3.4 PCL120-CHOL-TPrPr was synthesized and purified according to the procedure as in Example 2.3.1 above. A total of 86 mg of white polymer was obtained in 91% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 5.36, (s, 3H), 4.15 (dq, J=20.5, 8.4, 6.9 Hz, 238H), 3.63 (d, J=1.3 Hz, 448H), 3.33-3.22 (m, 99H), 2.91-2.46 (m, 810H), 2.12-1.81 (m, 163H), 1.76-1.35 (m, 983H), 1.13 (p, J=6.9 Hz, 349H), 0.95-0.82 (m, 382H), 0.66 (s, 6H).
9.3.5 Synthesis of PCL120-CHOL-TPrPr-ICG
[0276] Example 9.3.5 PCL120-CHOL-TPrPr-ICG was synthesized and purified according to the procedure as in Example 2.4.1 above. A total of 42 mg of green polymer was obtained in 76% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 8.06-7.30 (m, 18H), 5.36, (s, 3H), 4.13 (t, J=7.0 Hz, 230H), 3.63 (s, 448H), 3.27 (dq, J=11.8, 6.2, 5.3 Hz, 295H), 3.15-2.82 (m, 600H), 2.70 (s, 19H), 2.31 (d, J=8.7 Hz, 11H), 2.17-1.25 (m, 1693H), 0.98 (t, J=7.5 Hz, 621H), 0.66 (s, 14H).
Example 10
10. Example Series Containing C20H(CH.sub.2CH.sub.2OH):
10.1 PCL110-C2OH-TEPr-ICG
10.1.1 Synthesis Route of PCL110-C2OH-TEPr-ICG
##STR00077##
10.1.2 Synthesis of PCL110 (IB008-165)
[0277] Example 10.1.2 PCL110 was synthesized and purified according to the procedure as in Example 2.2.1 above (using 422 mg of 3-bromocaprolactone, 2.20 mmol) to give a total of 478 mg of white solid polymer in 89% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 4.26-4.10 (m, 331H), 3.63 (s, 448H), 2.16-1.95 (m, 253H), 1.80-1.38 (m, 482H).
10.1.3 Synthesis of PCL110-C2OH(IB015-003)
[0278] Example 10.1.3 PCL110-C2OH was synthesized and purified according to the procedure as in Example S1 above (using DMF and potassium carbonate) to obtain a total of 55 mg of white solid polymer in 93% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 4.26-4.10 (m, 316H), 3.63 (s, 448H), 3.42 (q, J=5.8 Hz, 30H), 3.22 (t, J=7.7 Hz, 15H), 2.65-2.56 (m, 44H), 2.16-1.95 (m, 253H), 1.80-1.38 (m, 482H).
10.1.4 Synthesis of PCL110-C2OH-TEPr (IB015-005)
[0279] Example 10.1.4 PCL110-C2OH-TEPr was synthesized and purified according to the procedure as in Example 2.3.1 above, to obtain a total of 62 mg of white polymer in 94% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 4.15 (dq, J=20.5, 8.4, 6.9 Hz, 218H), 3.63 (d, J=1.3 Hz, 448H), 3.42 (q, J=5.8 Hz, 30H), 3.33-3.22 (m, 114H), 2.74-2.46 (m, 406H), 2.40-3.35 (m, 376H), 2.12-1.81 (m, 163H), 1.76-1.35 (m, 953H), 1.13 (p, J=6.9 Hz, 349H), 0.95-0.82 (m, 382H).
10.1.5 Synthesis of PCL110-C2OH-TEPr-ICG (IB015-005)
[0280] Example 10.1.5 PCL110-C2OH-TEPr-ICG was synthesized and purified according to the procedure as in Example 2.4.1 above, to obtain a total of 52 mg of green polymer in 94% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 8.06-7.31 (m, 16H), 4.11 (q, J=7.2, 6.4 Hz, 223H), 3.63 (s, 448H), 3.22 (q, J=8.1 Hz, 105H), 2.79-2.54 (m, 389H), 2.37 (t, J=7.6 Hz, 331H), 1.96-1.81 (m, 133H), 1.69 (dt, J=14.9, 7.4 Hz, 369H), 1.61-1.34 (m, 624H), 1.13 (p, J=6.9 Hz, 349H), 0.95-0.82 (m, 382H).
10.2 PCL110-C2OH-TEPr-ICG
10.2.1 Synthesis Route of PCL110-C2OH-TEPr-ICG
##STR00078##
10.2.2 Synthesis of PCL110 (IB008-165)
[0281] Example 10.2.2 PCL110 was synthesized and purified according to the procedure as in Example 2.2.1 above (using 422 mg of 3-bromocaprolactone, 2.20 mmol) to give a total of 478 mg of white solid polymer in 89% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 4.26-4.10 (m, 331H), 3.63 (s, 448H), 2.16-1.95 (m, 253H), 1.80-1.38 (m, 482H).
10.2.3 PCL110-C2OH(IB015-004)
[0282] The synthesis and purification of Example 10.2.3 PCL110-C2OH was performed according to the procedure as in Example S1 above (using DMF and potassium carbonate)) to give a total of 58 mg of white solid polymer in 94% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 4.26-4.10 (m, 316H), 3.63 (s, 448H), 3.42 (q, J=5.8 Hz, 80H), 3.22 (t, J=7.7 Hz, 40H), 2.65-2.56 (m, 84H), 2.16-1.95 (m, 253H), 1.80-1.38 (m, 482H).
10.2.4 Synthesis of PCL110-C2OH-TEPr (IB015-006)
[0283] The synthesis and purification of Example 10.2.4 PCL110-C2OH-TEPr was carried out according to the procedure in Example 2.3.1 above, and a total of 57 mg of white polymer was obtained in 89% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 4.11 (q, J=7.2, 6.4 Hz, 223H), 3.63 (s, 448H), 3.22 (q, J=8.1 Hz, 105H), 2.79-2.54 (m, 389H), 2.37 (t, J=7.6 Hz, 331H), 1.69 (dt, J=14.9, 7.4 Hz, 369H), 1.43 (h, J=7.5 Hz, 527H), 1.16-1.13 (m, 224H), 0.95-0.82 (m, 214H).
10.2.5 Synthesis of PCL110-C2OH-TEPr-ICG (IB015-006)
[0284] The synthesis and purification of Example 10.2.5 PCL110-C2OH-TEPr-ICG was carried out according to the procedure as in Example 2.4.1 above, and a total of 42 mg of the green polymer was obtained in 81% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 8.06-7.31 (m, 20H), 4.11 (q, J=7.2, 6.4 Hz, 223H), 3.63 (s, 448H), 3.22 (q, J=8.1 Hz, 105H), 2.79-2.54 (m, 389H), 2.37 (t, J=7.6 Hz, 331H), 1.69 (dt, J=14.9, 7.4 Hz, 369H), 1.43 (h, J=7.5 Hz, 527H), 1.16-1.12 (m, 198H), 0.95-0.82 (m, 207H).
Example 11
11. Example series containing C2(-C2H.sub.5):
11.1 PCL110-C2-TPrPr-ICG
11.1.1 Synthesis Route of PCL110-C2-TPrPr-ICG
##STR00079##
11.1.2 Synthesis of PCL110 (IB008-165)
[0285] Example 11.1.2 PCL110 was synthesized and purified according to the procedure as in Example 2.2.1 above (using 422 mg of 3-bromocaprolactone, 2.20 mmol) to give a total of 478 mg of white solid polymer in 89% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 4.26-4.10 (m, 331H), 3.63 (s, 448H), 2.16-1.95 (m, 253H), 1.80-1.38 (m, 482H).
11.1.3 Synthesis of PCL110-C2 (IB008-171)
[0286] The synthesis and purification of Example 11.1.3 PCL110-C2 was performed according to the procedure as in Example S1 above (using DMF and potassium carbonate)) to give a total of 107 mg of white solid polymer in 94% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 4.28-4.01 (m, 282H), 3.63 (d, J=1.6 Hz, 448H), 3.22 (t, J=7.7 Hz, 38H), 2.61 (qd, J=7.3, 3.6 Hz, 77H), 2.06 (dq, J=25.1, 9.2, 7.9 Hz, 251H), 1.78-1.35 (m, 550H), 1.24 (t, J=7.4 Hz, 142H).
11.1.4 Synthesis of PCL110-C2-TPrPr (IB008-189)
[0287] Example 11.1.4 PCL110-C2-TPrPr was synthesized and purified according to the procedure as in Example 2.3.1 above, to give a total of 74.62 mg of white polymer in 91% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 4.09 (q, J=7.2, 6.6 Hz, 210H), 3.61 (s, 448H), 3.20 (q, J=8.1 Hz, 107H), 2.71-2.52 (m, 351H), 2.37 (q, J=8.9, 8.2 Hz, 369H), 1.86 (dd, J=14.6, 7.2 Hz, 117H), 1.66 (q, J=7.6 Hz, 328H), 1.43 (tq, J=15.1, 7.7 Hz, 629H), 1.21 (t, J=7.5 Hz, 188H), 0.84 (t, J=7.5 Hz, 557H).
11.1.5 Synthesis of PCL110-C2-TPrPr-ICG (IB008-189)
[0288] Example 11.1.5 PCL110-C2-TPrPr-ICG was synthesized and purified according to the procedure as in Example 2.4.1 above, to give a total of 62.7 mg of green polymer in 84% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 8.16-7.43 (m, 13H), 4.11 (q, J=7.4, 7.0 Hz, 239H), 3.63 (s, 448H), 3.22 (q, J=8.0 Hz, 111H), 2.75-2.55 (m, 385H), 2.38 (t, J=7.6 Hz, 277H), 1.88 (h, J=8.0 Hz, 146H), 1.67 (p, J=7.5 Hz, 379H), 1.45 (dp, J=14.9, 7.5, 6.7 Hz, 641H), 1.32-1.16 (m, 723H), 0.86 (t, J=7.4 Hz, 672H).
11.2 PCL103-C2-TPrPr-ICG
11.2.1 Synthesis Route of PCL103-C2-TPrPr-ICG
##STR00080##
11.2.2 Synthesis of PCL103 (IB008-099)
[0289] Example 11.2.2 PCL107 was synthesized and purified according to the procedure as in Example 2.2.1 above. A total of 465 mg of white solid polymer was obtained in 92% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 4.29-4.09 (m, 321H), 3.64 (s, 448H), 2.18-1.96 (m, 241H), 1.82-1.39 (m, 460H).
11.2.3 Synthesis of PCL103-C2 (IB008-147)
[0290] Synthesis and purification of Example 11.2.3 PCL103-C2 were performed according to the procedure as in Example S1 above (using DMF and potassium carbonate), a total of 43 mg of the product was obtained as a white solid polymer in 92% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 4.12 (q, J=6.8 Hz, 233H), 3.64 (s, 448H), 3.22 (t, J=7.8 Hz, 61H), 2.62 (dp, J=12.3, 7.2, 5.1 Hz, 146H), 1.89 (q, J=7.5 Hz, 144H), 1.76-1.62 (m, 197H), 1.60-1.34 (m, 362H), 1.25 (d, J=7.1 Hz, 299H).
11.2.4 Synthesis of PCL103-C2-TPrPr (IB008-147)
[0291] Example 11.2.4 PCL103-C2-TPrPr was synthesized and purified according to the procedure in Example 2.3.1 above, and a total of 51 mg of the product was obtained as white polymer in 89% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 4.12 (q, J=6.8 Hz, 203H), 3.64 (s, 448H), 3.22 (t, J=7.8 Hz, 95H), 2.62 (dp, J=12.3, 7.2, 5.1 Hz, 247H), 2.39 (t, J=7.7 Hz, 111H), 1.89 (q, J=7.5 Hz, 144H), 1.76-1.62 (m, 321H), 1.60-1.34 (m, 362H), 1.25 (d, J=7.1 Hz, 299H), 0.87 (t, J=7.4 Hz, 188H).
11.2.5 Synthesis of PCL103-C2-TPrPr-ICG (IB008-147)
[0292] The synthesis and purification of Example 11.2.5 PCL103-C2-TPrPr-ICG was carried out according to the procedure in Example 2.4.1 above, and a total of 43 mg of green polymer was obtained in 83% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 8.13-7.31 (m, 17H), 4.11 (t, J=6.7 Hz, 203H), 3.64 (s, 448H), 3.22 (t, J=7.9 Hz, 95H), 2.62 (h, J=6.9, 5.1 Hz, 247H), 2.39 (t, J=7.7 Hz, 111H), 1.96-1.81 (m, 144H), 1.68 (p, J=7.3 Hz, 321H), 1.60-1.34 (m, 362H), 1.30-1.17 (m, 299H), 0.87 (t, J=7.4 Hz, 188H).
11.3 PCL110-C2-TPrPr-ICG
11.3.1 Synthesis Route of PCL110-C2-TPrPr-ICG
##STR00081##
11.3.2 Synthesis of PCL110 (IB008-165)
[0293] Example 11.3.2 PCL110 was synthesized and purified according to the procedure as in Example 2.2.1 above (using 422 mg of 3-bromocaprolactone, 2.20 mmol) to give a total of 478 mg of white solid polymer in 89% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 4.26-4.10 (m, 331H), 3.63 (s, 448H), 2.16-1.95 (m, 253H), 1.80-1.38 (m, 482H).
11.3.3 Synthesis of PCL110-C2 (IB008-169)
[0294] Synthesis and purification of Example 11.3.3 PCL110-C2 were performed according to the procedure as in Example S1 above (using DMF and potassium carbonate), a total of 43 mg of the product was obtained as a white solid polymer in 92% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 4.17 (dtd, J=21.8, 11.6, 9.8, 6.4 Hz, 307H), 3.63 (d, J=1.7 Hz, 448H), 3.22 (t, J=7.6 Hz, 12H), 2.61 (dq, J=10.6, 6.2 Hz, 25H), 2.05 (tdd, J=23.3, 16.9, 11.3 Hz, 215H), 1.78-1.37 (m, 503H), 1.28-1.19 (m, 62H).
11.3.4 Synthesis of PCL110-C2-TPrPr (IB008-187)
[0295] Example 11.3.4 PCL110-C2-TPrPr was synthesized and purified according to the procedure in Example 2.3.1 above, and a total of 51 mg of the product was obtained as white polymer in 89% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 4.04 (dt, J=9.2, 5.1 Hz, 210H), 3.56 (d, J=2.2 Hz, 448H), 3.16 (t, J=7.6 Hz, 101H), 2.65-2.46 (m, 456H), 2.31 (q, J=8.0 Hz, 530H), 1.81 (d, J=9.8 Hz, 117H), 1.61 (q, J=7.7 Hz, 327H), 1.35 (h, J=7.4 Hz, 876H), 1.17 (q, J=7.8 Hz, 87H), 0.79 (t, J=7.4 Hz, 801H).
11.3.5 Synthesis of PCL110-C2-TPrPr-ICG (IB008-187)
[0296] The synthesis and purification of Example 11.3.5 PCL110-C2-TPrPr-ICG was carried out according to the procedure in Example 2.4.1 above, and a total of 43 mg of green polymer was obtained in 83% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 8.13-7.31 (m, 21H) 4.04 (t, J=9.2, 5.1 Hz, 210H), 3.56 (d, J=2.2 Hz, 448H), 3.16 (t, J=7.6 Hz, 101H), 2.65-2.46 (m, 456H), 2.31 (q, J=8.0 Hz, 530H), 1.81 (d, J=9.8 Hz, 117H), 1.61 (q, J=7.7 Hz, 325H), 1.35 (h, J=7.4 Hz, 856H), 1.17 (q, J=7.8 Hz, 83H), 0.79 (t, J=7.4 Hz, 805H).
Example 12
12. Example Series Containing Amphoteric Ions:
12.1 PCL110-EtriMAEP-TPrPr-ICG
12.1.1 Synthesis Route of PCL110-EtriMAEP-TPrPr-ICG
##STR00082##
12.1.2 Synthesis of PCL110 (IB008-165)
[0297] Example 12.1.2 PCL110 was synthesized and purified according to the procedure as in Example 2.2.1 above (using 422 mg of 3-bromocaprolactone, 2.20 mmol) to give a total of 478 mg of white solid polymer in 89% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 4.26-4.10 (m, 331H), 3.63 (s, 448H), 2.16-1.95 (m, 253H), 1.80-1.38 (m, 482H).
12.1.3 Synthesis of PCL110-EtriMAEP (IB015-008)
[0298] Synthesis and purification of Example 12.1.3 PCL110-EtriMAEP were performed according to the procedure as in Example S1 above (using DMF and potassium carbonate), a total of 120 mg of product was obtained as a white solid polymer in 94% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 4.33-4.12 (m, 351H), 4.02-3.95 (m, 40H), 3.68-3.63 (m, 488H), 3.25-3.18 (m, 220H), 2.16-1.95 (m, 253H), 1.80-1.38 (m, 482H).
12.1.4 Synthesis of PCL110-EtriMAEP-TPrPr (IB015-013)
[0299] Example 12.1.4 PCL110-EtriMAEP-TPrPr was synthesized and purified according to the procedure as in Example 2.3.1 above, yielding a total of 65 mg of the product as a white polymer in 89% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 4.33-4.12 (m, 351H), 4.02-3.95 (m, 40H), 3.68-3.63 (m, 488H), 3.25-3.18 (m, 180H), 2.63-2.51 (m, 40H), 2.14-1.98 (m, 253H), 1.80-1.38 (m, 482H).
12.1.5 Synthesis of PCL110-EtriMAEP-TPrPr-ICG (IB015-013)
[0300] The synthesis and purification of Example 12.1.5 PCL110-EtriMAEP-TPrPr-ICG were performed according to the procedure as in Example 2.4.1 above, yielding a total of 47.9 mg of green polymer in 81% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 8.06-7.31 (m, 25H), 4.33-4.12 (m, 351H), 4.02-3.95 (m, 42H), 3.68-3.63 (m, 488H), 3.25-3.18 (m, 196H), 2.71-2.51 (m, 411H), 2.40-2.36 (m, 353), 2.09-2.14-1.98 (m, 253H), 1.80-1.38 (m, 852H), 0.89-0.85 (m, 528H).
12.2 PCL110-EMMPS-TPrPr-ICG
12.2.1 Synthesis Route of PCL110-EMMPS-TPrPr-ICG
##STR00083##
12.2.2 Synthesis of PCL110 (IB008-165)
[0301] The synthesis and purification of example 12.2.2 PCL110 were performed according to the procedure in example 2.2.1 above (using 422 mg 3-bromocaprolactone, 2.20 mmol) to obtain a total of 478 mg white solid polymer with a yield of 89%. .sup.1H NMR (400 MHZ, Chloroform-d) 4.26-4.10 (m, 331H), 3.63 (s, 448H), 2.16-1.95 (m, 253H), 1.80-1.38 (m, 482H).
12.2.3 Synthesis of PCL110-EMMPS (IB015-014)
[0302] Synthesis and purification of Example 12.2.3 PCL110-EMMPS were performed according to the procedure as in Example S1 above (using DMF and potassium carbonate), a total of 115 mg of product was obtained as a white solid polymer in 93% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 4.26-4.10 (m, 311H), 3.63 (s, 448H), 3.47-3.32 (m, 82H), 3.28-3.18 (m, 20H), 3.00 (s, 120H), 2.90-2.76 (m, 87H), 2.17-1.95 (m, 294H), 1.80-1.38 (m, 482H).
12.2.4 Synthesis of PCL110-EMMPS-TPrPr (IB015-014)
[0303] Example 12.2.4 PCL110-EMMPS-TPrPr was synthesized and purified according to the procedure as in Example 2.3.1 above, yielding a total of 65 mg of the product as a white polymer in 89% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 4.26-4.10 (m, 311H), 3.63 (s, 448H), 3.47-3.32 (m, 82H), 3.28-3.18 (m, 20H), 3.00 (s, 120H), 2.90-2.76 (m, 87H), 2.70-2.53 (m, 352H), 2.40-2.60 (m, 348H), 2.17-1.95 (m, 294H), 1.80-1.38 (m, 838H), 0.89-0.85 (m, 588H).
12.2.5 Synthesis of PCL110-EMMPS-TPrPr-ICG
[0304] Example 12.2.5 PCL110-EMMPS-TPrPr-ICG was synthesized and purified according to the procedure as in Example 2.4.1 above, yielding a total of 49.9 mg of the green polymer in 82% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 8.06-7.31 (m, 30H) 4.26-4.10 (m, 300H), 3.63 (s, 448H), 3.47-3.32 (m, 79H), 3.28-3.18 (m, 22H), 3.00 (s, 119H), 2.90-2.76 (m, 89H), 2.70-2.53 (m, 332H), 2.40-2.60 (m, 352H), 2.17-1.95 (m, 292H), 1.80-1.38 (m, 886H), 0.89-0.85 (m, 571H).
12.3 PCL110-EMMPC-TPrPr-ICG
12.3.1 Synthesis Route of PCL110-EMMPC-TPrPr-ICG
##STR00084##
12.3.2 Synthesis of PCL110
[0305] The synthesis and purification of example 12.3.2 PCL110 were performed according to the procedure in example 2.2.1 above (using 422 mg 3-bromocaprolactone, 2.20 mmol) to obtain a total of 478 mg white solid polymer with a yield of 89%. .sup.1H NMR (400 MHZ, Chloroform-d) 4.26-4.10 (m, 331H), 3.63 (s, 448H), 2.16-1.95 (m, 253H), 1.80-1.38 (m, 482H).
12.3.3 Synthesis of PCL110-EMMA
[0306] Synthesis and purification of Example 12.2.3 PCL110-EMMA were performed according to the procedure as in Example S1 above (using DMF and potassium carbonate), a total of 123 mg of the product was obtained as a white solid polymer in 94% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 4.29 (s, 39H), 4.26-4.10 (m, 311H), 3.81-3.77 (m, 41H), 3.63 (s, 448H), 3.21-3.18 (m, 19H), 2.96-2.92 (m, 40H), 2.16-1.95 (m, 253H), 1.80-1.38 (m, 482H).
12.3.4 Synthesis of PCL110-EMMA-TPrPr
[0307] Example 12.3.4 PCL110-EMMA-TPrPr was synthesized and purified according to the procedure as in Example 2.3.1 above, yielding a total of 65 mg of the product as a white polymer in 83% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 4.29 (s, 39H), 4.26-4.10 (m, 311H), 3.81-3.77 (m, 41H), 3.63 (s, 448H), 3.21-3.18 (m, 19H), 2.96-2.92 (m, 40H), 2.70-2.53 (m, 358H), 2.40-2.36 (m, 365H), 2.16-1.95 (m, 243H), 1.80-1.38 (m, 872H). 0.89-0.85 (m, 605H).
12.3.5 Synthesis of PCL110-EMMA-TPrPr-ICG
[0308] Example 12.3.5 PCL110-EMMA-TPrPr-ICG was synthesized and purified according to the procedure as in Example 2.4.1 above, yielding a total of 40.5 mg of the green polymer in 79% yield. .sup.1H NMR (400 MHz, Chloroform-d) 8.11-7.28 (m, 25H), 4.29 (s, 39H), 4.26-4.10 (m, 311H), 3.81-3.77 (m, 41H), 3.63 (s, 448H), 3.21-3.18 (m, 19H), 2.96-2.92 (m, 40H), 2.70-2.53 (m, 358H), 2.40-2.36 (m, 361H), 2.16-1.95 (m, 243H), 1.80-1.38 (m, 911H). 0.89-0.85 (m, 610H).
Example 13
13. Example Series Containing C4(-C.sub.4H.sub.9):
13.1 PCL110-C4-TPrPr-ICG
13.1.1 Synthesis Route of PCL110-C4-TPrPr-ICG
##STR00085##
13.1.2 Synthesis of PCL110 (IB008-165)
[0309] The synthesis and purification of Example 13.1.2 PCL110 were performed according to the procedure in example 2.2.1 above (using 422 mg 3-bromocaprolactone, 2.20 mmol) to obtain a total of 478 mg white solid polymer with a yield of 89%. .sup.1H NMR (400 MHZ, Chloroform-d) 4.26-4.10 (m, 331H), 3.63 (s, 448H), 2.16-1.95 (m, 253H), 1.80-1.38 (m, 482H).
13.1.3 Synthesis of PCL110-C4 (IB015-017)
[0310] Synthesis and purification of Example 13.1.3 PCL110-C4 were performed according to the procedure as in Example S1 above (using DMF and potassium carbonate), a total of 107 mg of the product was obtained as a white solid polymer in 94% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 4.28-4.01 (m, 291H), 3.63 (d, J=1.6 Hz, 448H), 3.22 (t, J=7.7 Hz, 40H), 2.61 (qd, J=7.3, 3.6 Hz, 77H), 2.06 (dq, J=25.1, 9.2, 7.9 Hz, 251H), 1.78-1.35 (m, 630H), 1.24 (t, J=7.4 Hz, 142H).
13.1.4 Synthesis of PCL110-C4-TPrPr (IB015-018)
[0311] Example 13.1.4 PCL110-C4-TPrPr was synthesized and purified according to the procedure as in Example 2.3.1 above, yielding a total of 74.62 mg of the product as a white polymer in 91% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 4.09 (q, J=7.2, 6.6 Hz, 210H), 3.61 (s, 448H), 3.20 (q, J=8.1 Hz, 110H), 2.71-2.52 (m, 351H), 2.37 (q, J=8.9, 8.2 Hz, 369H), 1.86 (dd, J=14.6, 7.2 Hz, 120H), 1.66 (q, J=7.6 Hz, 328H), 1.43 (tq, J=15.1, 7.7 Hz, 711H), 1.21 (t, J=7.5 Hz, 190H), 0.84 (t, J=7.5 Hz, 582H).
13.1.5 Synthesis of PCL110-C4-TPrPr-ICG (IB015-018)
[0312] Example 13.1.5 PCL110-C4-TPrPr-ICG was synthesized and purified according to the procedure as in Example 2.4.1 above, and a total of 62.7 mg of the green polymer was obtained in 84% yield. .sup.1H NMR (400 MHz, Chloroform-d) 8.16-7.43 (m, 25H), 4.11 (q, J=7.4, 7.0 Hz, 239H), 3.63 (s, 448H), 3.22 (q, J=8.0 Hz, 125H), 2.75-2.55 (m, 375H), 2.38 (t, J=7.6 Hz, 287H), 1.88 (h, J=8.0 Hz, 147H), 1.67 (p, J=7.5 Hz, 380H), 1.45 (dp, J=14.9, 7.5, 6.7 Hz, 641H), 1.32-1.16 (m, 804H), 0.86 (t, J=7.4 Hz, 672H).
Example 14
14. Example Series Containing Delivery Molecules:
14.1 PCL110-5ALA-TPrPr-ICG
14.1.1 Synthesis Route of PCL110-5ALA-TPrPr-ICG
##STR00086##
14.1.2 Synthesis of PCL110 (IB008-165)
[0313] The synthesis and purification of Example 14.1.2 PCL110 were performed according to the procedure in example 2.2.1 above (using 422 mg 3-bromocaprolactone, 2.20 mmol) to obtain a total of 478 mg white solid polymer with a yield of 89%. .sup.1H NMR (400 MHZ, Chloroform-d) 4.26-4.10 (m, 331H), 3.63 (s, 448H), 2.16-1.95 (m, 253H), 1.80-1.38 (m, 482H).
14.1.3 Synthesis of PCL110-5ALA (IB007-177)
[0314] Synthesis and purification of Example 14.1.3 PCL110-5ALA were performed according to the procedure as in Example S1 above (using DMF and potassium carbonate), a total of 120 mg of product was obtained as a white solid polymer in 94% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 4.30-3.98 (m, 287H), 3.64 (s, 448H), 3.20 (m 27H), 2.71 (d, J=6.6 Hz, 57H), 2.60 (dt, J=25.0, 7.2 Hz, 123H), 2.04 (s, 167H), 1.82-1.71 (m, 188H), 1.60 (q, J=6.7 Hz, 309H), 1.41-1.22 (m, 254H).
14.1.4 Synthesis of PCL110-5ALA-TPrPr (IB008-191)
[0315] Example 14.1.4 PCL110-5ALA-TPrPr was synthesized and purified according to the procedure as in Example 2.3.1 above, yielding a total of 35 mg of white polymer in 82% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 4.16 (s, 228H), 3.64 (s, 448H), 3.47-2.96 (m, 306H), 2.58 (d, J=8.1 Hz, 70H), 2.18 (s, 231H), 1.87 (s, 258H), 1.56 (s, 202H), 1.49-1.18 (m, 452H), 1.03 (s, 180H), 0.88 (t, J=6.7 Hz, 63H).
14.1.5 Synthesis of PCL110-5ALA-TPrPr-ICG (IB008-191)
[0316] Example 14.1.5 PCL110-5ALA-TPrPr-ICG was synthesized and purified according to the procedure as in Example 2.4.1 above, and a total of 22 mg of the green polymer was obtained in 72% yield. .sup.1H NMR (400 MHZ, Chloroform-d) 8.46-7.34 (m, 11H) 4.30-3.98 (m, 382H), 3.64 (s, 448H), 3.24 (d, J=35.8 Hz, 135H), 3.05 (s, 188H), 2.75-2.50 (m, 183H), 2.14-1.21 (m, 1372H), 1.01 (t, J=7.4 Hz, 196H), 0.86 (s, 49H).
Example 15
15.1 Tumor-Bearing Mice Fluorescence Imaging Experiment for C9H19-Containing Polymer Imaging Probes
[0317] Animal Model: Female Balb/c nude mice (4-6 weeks age) are inoculated with 4T1 cells (app. 210.sup.6/mouse) into the right flank. Imaging Experiment is conducted on mice when tumor volume reaches 200-400 mm.sup.3.
[0318] Dose of Imaging Probe: Imaging Probe (IB008-103) was injected via tail vein at 2.5 mg/kg.
[0319] in vivo Fluorescence Imaging: A fluorescence imaging system (PerkinElmer, IVIS spectrum CT, Made: U.S.A, each fluorescence imaging capture uses the system-default filters for ICG with identical imaging parameters) is used to observe the in vivo biodistribution and tumor accumulation. As exhibited in
15.2 Measurement of the Ratio of Tumor to Healthy Tissue (Muscle) after Administration of Imaging Agent (IB008-103, Structure Corresponds to Example 7.1.4)
[0320] At different time points (5 min, 30 min, 1 hour, 2 hour, 4 hour, 8 hour, 16 hour, 24 hour, 48 hour, 96 hour, 144 hour, 196 hour after administration), mice are sacrificed to harvest organs for ex vivo fluorescence imaging measurement. Organs collected include: Heart, Liver, Spleen, Lung, Kidney, Tumor, Muscle. Blood is also collected and centrifuged into plasma. After tissue imaging, identical area for Region of Interest (ROI) was used in imaging system software to compare the fluorescence intensity (total intensity and average intensity). Typical data is shown in
[0321] In
15.3 Biodistribution and Dose Comparison
[0322] Three doses (2.5 mg/kg, 5.0 mg/kg, 10.0 mg/kg) were compared using an polymer imaging probe (IB008-103). A total of 6 4T1 tumor-bearing mice were use for each dose (3 of them were sacrificed at 8 hours post injection, the other 3 of them were sacrificed at 24 hours post injection).
15.4 Pharmacokinetics and Blood Stream Half Life
[0323] As described in 15.2, upon sacrifice of each animal, about 1 ml of blood is collected. After fluorescence imaging, the blood is centrifuged (4 C., 8000 rpm, 5 minute) within 1 hour after isolation to generate plasma. Using a fluorospectrophotometer (F97, Lengguang Tech, Shanghai), the plasma is measured for fluorescence intensity (780 nm excitement and 820 nm peak value as measured intensity). Then following in-house standard procedure to acidify the plasma, then the acidified plasma is measure for fluorescence intensity. The fluorescence intensity of the acidified plasma is proportional to the fluorophore (in this case, ICG) in the blood. The result is plotted in
Example 16
16.1 Tumor-Bearing Mice Fluorescence Imaging Experiment for Polymer Imaging Probes
[0324] Animal Model: Female Balb/c nude mice (4-6 weeks age) are inoculated with 4T1 cells (app. 210.sup.6/mouse) into the mammary pad. Imaging Experiment is conducted on mice when tumor volume reaches 200-400 mm.sup.3.
[0325] Dose of Imaging Probe: Imaging Probe (IB008-103) was injected via tail vein at 2.5 mg/kg. in vivo Fluorescence Imaging: A fluorescence imaging system (PerkinElmer, IVIS spectrum CT, Made: U.S.A, each fluorescence imaging capture uses the system-default filters for ICG with identical imaging parameters) is used to observe the in vivo biodistribution and tumor accumulation. As exhibited in
Example 17
17.1 Tumor-Bearing Mice Fluorescence Imaging Experiment for Cholesterol-Containing Polymer Imaging Probes
[0326] Animal Model: Female Balb/c nude mice (4-6 weeks age) are inoculated with 4T1 cells (app. 210.sup.6/mouse) into the mammary pad. Imaging Experiment is conducted on mice when tumor volume reaches 200-400 mm.sup.3.
[0327] Dose of Imaging Probe: Imaging Probe (IB008-027, example 9.3.5) was injected via tail vein at 2.5 mg/kg.
[0328] in vivo Fluorescence Imaging: A fluorescence imaging system (PerkinElmer, IVIS spectrum CT, Made: U.S.A, each fluorescence imaging capture uses the system-default filters for ICG with identical imaging parameters) is used to observe the in vivo biodistribution and tumor accumulation. As exhibited in
Example 18
18.1 Tumor-Bearing Mice Fluorescence Imaging Experiment for Polymer Imaging Probes Containing Some Hydrophilic and Hydrophobic Groups
[0329] Animal Model: Female Balb/c nude mice (4-6 weeks age) are inoculated with 4T1 breast cancer cells (app. 210.sup.6/mouse) subcutaneously. Imaging Experiment is conducted on mice when tumor volume reaches 200-400 mm.sup.3.
[0330] Dose of Imaging Probe: Imaging Probes (IB008-139, example 8.1.4 corresponding to C18H.sub.37-hydrophobic group; IB008-147, example 11.2.1 corresponding to C2H.sub.5-hydrophobic group; and IB015-014, example 12.2.1 corresponding to zwitterionic EMMPS-group) were injected via tail vein at 2.5 mg/kg.
[0331] in vivo Fluorescence Imaging: A fluorescence imaging system (PerkinElmer, IVIS spectrum CT, Made: U.S.A, each fluorescence imaging capture uses the system-default filters for ICG with identical imaging parameters) is used to observe the in vivo bio-distribution and tumor accumulation. As exhibited in
Example 19
19.1 Tumor-Bearing Mice Fluorescence Imaging Experiment for Polymer Imaging Probes
[0332] Animal Model: Female Balb/c nude mice (4-6 weeks age) are inoculated with 4T1 cells (app. 210.sup.6/mouse) into the mammary pad. Imaging Experiment is conducted on mice when tumor volume reaches 200-400 mm.sup.3.
[0333] Dose of Imaging Probe: Imaging Probe (IB015-018, example 13.1.1 corresponding to C4H.sub.9-group) was injected via tail vein at 2.5 mg/kg.
[0334] in vivo Fluorescence Imaging: A fluorescence imaging system (PerkinElmer, IVIS spectrum CT, Made: U.S.A, each fluorescence imaging capture uses the system-default filters for ICG with identical imaging parameters) is used to observe the in vivo bio-distribution and tumor accumulation. As exhibited in
Example 20
20.1 Pathological Assessment of Harvested Lymph Nodes with Higher Fluorescence Intensity
[0335] Lymph nodes of higher fluorescence intensity obtained in Example 19 are placed in formaldehyde solution within 3 minutes after being harvested. Standardized wax embedding was prepared to generate series adjacent tissue slices of 2-8 microns thick. Once slice was used to prepare HE staining, and another adjacent slice was used to conduct fluorescence label marking with markers specific to panCK (marking cancer cells) and DAPI (marking the nucleus). The images are shown in
Example 21
21.1 Tumor-bearing Mice Fluorescence Imaging Experiment for Polymer Imaging Probes Containing 5ALA and ICG (Dual Wavelength)
[0336] Animal Model: Female Balb/c nude mice (4-6 weeks age) are inoculated with 4T1 cells (app. 210.sup.6/mouse) into the subcutaneously. Imaging Experiment is conducted on mice when tumor volume reaches 200-400 mm.sup.3.
[0337] Dose of Imaging Probe: Imaging Probe (IB008-191, example 14.1.1 corresponding to 5ALA and ICG containing polymer) was injected via tail vein at 2.5 mg/kg.
[0338] in vivo Fluorescence Imaging: A fluorescence imaging system (PerkinElmer, IVIS spectrum CT, Made: U.S.A, each fluorescence imaging capture uses the system-default filters for ICG with identical imaging parameters) is used to observe the in vivo biodistribution and tumor accumulation. For 5ALA, the exciting wavelength is 465 nm, and the emission filters of 620 nm, 640 nm, and 680 nm were tried. As exhibited in
Example 22 Fluorescence Intensity Measurement for Polymer Nano-Particle Solutions
22.1 Preparation of Polymer Nano-Particle Solutions (1 mg/ml, Similar to Procedure Described in Example 6.1)
22.1 Fluorescence Intensity Measurement
[0339] 100 L stock solution of polymer nano-particle solution (1 mg/ml, prepared according to Example 6.1) was diluted to 2.0 mL PBS or Citric Acid buffer solutions. After mixing, fluorescence intensity was measured using an exciting wavelength of 730 nm, and the emission detection range was set for 785-900 nm scan.
[0340] Fmax (maximum fluorescence intensity) is defined as the maximum peak intensity measured for the entire pH range (reading the peak value at about 820 nm, subtracting background value).
[0341] Fmin (minimum fluorescence intensity) is defined as the minimum peak intensity measured for the entire pH range (reading the peak value at about 820 nm, subtracting background value).
[0342] The Ratio of Fmax/Fmin is calculated numerically using Fmax and Fmin values.
[0343] pHt (pH transition point) is determined by conducting a normalization treatment using 821 nm peak intensity values for solutions of different pH values. Then a Boltzmann fitting was employed to fit the data. The pH value corresponding to the 50% value of the highest intensity data was taken as pHt.
[0344] An assessment of the magnitude of fluorescence change due to pH change is assessed by a parameter defined as pH.sub.50%. Using above-mentioned normalization data plot, pH.sub.10% is the pH value at 10% value of the highest intensity data, pH.sub.50% is the pH value at 50% value of the highest intensity data, pH.sub.90% is the pH value at 90% value of the highest intensity data.
[0345] All data are summarized in
[0346] In summary, this invention effectively overcomes various shortcomings in the prior art and has a high industrial value.
[0347] The foregoing examples only exemplarily illustrate the principles and effects of the present invention, and are not used to limit the present invention. Anyone familiar with this technology can modify or change the above-mentioned examples without departing from the spirit and scope of this invention. Therefore, all equivalent modifications or changes completed by those with ordinary knowledge in the technical field without departing from the spirit and technical ideas disclosed in this invention should still be covered by the claims of this invention.