Albumin-binding prodrug, albumin-drug conjugate, method for preparing albumin-drug conjugate, and pharmaceutical composition for preventing or treating cancer comprising same

Abstract

Disclosed are: an albumin-binding prodrug that can stably and selectively bind to albumin capable of targeting cancer while also stably binding to drugs; an albumin-drug conjugate in which the albumin-binding prodrug is bound to albumin; a method for preparing the albumin-drug conjugate; and a pharmaceutical composition for preventing or treating cancer, comprising the albumin-binding prodrug or the albumin-drug conjugate.

Description

Albumin-binding prodrug, albumin-drug conjugate, method for preparing albumin-drug conjugate, and pharmaceutical composition for preventing or treating cancer comprising the same

[0001] The present invention relates to an albumin-binding prodrug, an albumin-drug conjugate, a method for preparing an albumin-drug conjugate, and a pharmaceutical composition for preventing or treating cancer comprising the same.

[0002] Enhanced Permeability and Retention (EPR) is the theory that nanoparticle-sized substances can accumulate in cancer cells due to the loose blood vessels that develop around them. Albumin Binding Proteins (ABP) is the theory that cancer cells obtain necessary nutrients from albumin in the blood.

[0003] Albumin is abundant in blood and plays a role in transporting various nutrients, including lipids and metal ions. It is known that blood albumin levels are reduced in cancer patients. Considering this experience and the aforementioned EPR and ABP effects, there is a growing demand for conjugates that link anticancer drugs to albumin.

[0004] Abraxane is a drug made of albumin nanoparticles loaded with the anticancer drug paclitaxel. It has received FDA clinical approval for use in cancer treatment. However, because the binding between albumin nanoparticles and paclitaxel is relatively weak, the drug is rapidly released into the bloodstream, causing toxicity in normal cells.

[0005] Aldoxorubicin, a pharmaceutical product that binds the anticancer drug doxorubicin to the cysteine-34 thiol group protruding from the albumin surface via a maleimide linker, is currently in clinical use. The problem with aldoxorubicin is that the protruding thiol group can cause albumin to degrade due to an immune response.

[0006] In this way, it is difficult to stably combine anticancer drugs and albumin simultaneously, and considering various variables, it is necessary to verify whether the conjugate of anticancer drugs and albumin will actually exhibit a cancer treatment effect.

[0007] The purpose of the present disclosure is to provide a conjugate or a precursor thereof that stably binds a drug such as an anticancer agent to albumin simultaneously, thereby allowing the drug to selectively accumulate in a cancer tumor, thereby improving the therapeutic effect and minimizing side effects of anticancer treatment.

[0008] However, the purpose of the present disclosure is not limited to the tasks mentioned above, and other tasks not mentioned will be clearly understood by those skilled in the art from the description below.

[0009] According to one aspect, an albumin-binding prodrug represented by the following chemical formula I is disclosed:

[0010] <Chemical Formula I>

[0011] Ac-(CL1) c1 -(SL1) s1 -(CL2) c2 -(SL2) s2 -D

[0012] In the above chemical formula I,

[0013] Ac is an albumin-binding moiety represented by the following chemical formula 1,

[0014] CL1 and CL2 are each a cleavable linker,

[0015] SL1 and SL2 are each soluble linkers,

[0016] c1, c2, s1 and s2 are each 0 or 1,

[0017] D is a drug moiety,

[0018] <Chemical Formula 1>

[0019]

[0020] In the above chemical formula 1,

[0021] Z is N, O or C(R 14 ) and,

[0022] R 11 Inland R 14 , R 21 Inland R 24 And R3 to R6 are each independently hydrogen, deuterium, -F, -Cl, -Br, -I, hydroxyl group, cyano group, nitro group, formyl group, carboxyl group, amino group, substituted or unsubstituted C1-C 10 Alkyl group, substituted or unsubstituted C2-C 10 Alkenyl group, substituted or unsubstituted C2-C 10 Alkynyl group, substituted or unsubstituted C1-C 10 Alkoxy group, substituted or unsubstituted C6-C 10 Aryl group, -N(Q1)(Q2) and -C(=O)(Q1), selected from

[0023] R 11 Inland R 14 and R 21 Inland R 24 One of them is (CL1) in the above chemical formula I c1 -(SL1) s1 -(CL2) c2 -(SL2) s2 -This is a site that combines with D,

[0024] Optionally i) R 11 and R 12 , ii) R 12 Wow R 13 , iii) R 13 and R 14 , iv) R 21 and R 22 , v) R 22 Wow R 23 , vi) R 23 and R 24 , or vii) any combination thereof, which may be combined with each other to form a substituted or unsubstituted C3-C 10 Carbocyclic group or substituted or unsubstituted C1-C 10Forming a heterocyclic group,

[0025] Q1 and Q2 are independently hydrogen, deuterium, -F, -Cl, -Br, -I, hydroxyl group, cyano group, nitro group, formyl group, carboxyl group, amino group, substituted or unsubstituted C1-C 10 Alkyl group, substituted or unsubstituted C2-C 10 Alkenyl group, substituted or unsubstituted C2-C 10 Alkynyl group, substituted or unsubstituted C1-C 10 Alkoxy group and substituted or unsubstituted C6-C 10 Selected from aryl groups,

[0026] substituted C1-C 10 Alkyl group, substituted C2-C 10 Alkenyl group, substituted C2-C 10 alkynyl group, substituted C1-C 10 Alkoxy group, substituted C6-C 10 Aryl group, substituted C3-C 10 Carbocyclic groups and substituted C1-C 10 The substituents of the heterocyclic group are deuterium, -F, -Cl, -Br, -I, hydroxyl group, cyano group, nitro group, formyl group, carboxyl group, amino group, C1-C 10 Alkyl group and C6-C 10 Selected from aryl groups,

[0027] m is an integer selected from 1 to 5.

[0028] According to one implementation example, R 11 Inland R 14 One of the above chemical formulas (CL1) c1 -(SL1) s1 -(CL2) c2 -(SL2) s2 -It may be a site that is combined with D.

[0029] According to one implementation example, R 21 Inland R 24 One of the above chemical formulas (CL1) c1 -(SL1) s1-(CL2) c2 -(SL2) s2 - It may be a site that is combined with D. For example, R 22 or R 23 Among the above chemical formula I (CL1) c1 -(SL1) s1 -(CL2) c2 -(SL2) s2 -It may be a site that is combined with D.

[0030] According to one embodiment, Z in the above chemical formula 1 is C(R 14 ) may be.

[0031] According to one embodiment, R in the chemical formula 1 11 Inland R 14 are independently hydrogen, deuterium, -F, -Cl, -Br, -I, hydroxyl group, cyano group, nitro group, formyl group, carboxyl group, amino group, substituted or unsubstituted C1-C 10 Alkyl group, substituted or unsubstituted C2-C 10 Alkenyl group, substituted or unsubstituted C1-C 10 Alkoxy groups can be selected from -N(Q1)(Q2) and -C(=O)(Q1).

[0032] According to one implementation example, R 12 Wow R 13 are combined with each other to form substituted or unsubstituted C3-C 10 Carbocyclic group or substituted or unsubstituted C1-C 10 It can form a heterocyclic group. That is, it can form a condensed ring that is condensed with the Z-containing ring in the above chemical formula 1.

[0033] According to one embodiment, R in the chemical formula 1 21 Inland R 24 is (CL1) in the above chemical formula I c1 -(SL1) s1 -(CL2) c2 -(SL2) s2-The site bonded to -D can be selected from hydrogen, deuterium, -F, -Cl, -Br, -I, hydroxyl group, cyano group, nitro group, formyl group, carboxyl group, amino group, -N(Q1)(Q2) and -C(=O)(Q1).

[0034] Among the above-described -N(Q1)(Q2) and -C(=O)(Q1), Q1 and Q2 may independently be hydrogen, deuterium, a hydroxyl group, a formyl group, a carboxyl group, an amino group, a methyl group, an ethyl group, or a phenyl group.

[0035] According to one embodiment, R3 in the above chemical formula 1 may be hydrogen, deuterium, -F, -Cl, -Br, -I, a hydroxyl group or a phenyl group.

[0036] According to one embodiment, R4 in the above chemical formula 1 may be a hydroxyl group.

[0037] According to one embodiment, R5 and R6 in the above chemical formula 1 may be hydrogen, deuterium, a methyl group or a phenyl group.

[0038] According to one embodiment, m in the chemical formula 1 may be 1 or 2.

[0039] According to one embodiment, Ac in the above formula I may be an albumin-binding moiety represented by any one of the following formulae 1-1 to 1-6:

[0040] <Chemical Formula 1-1>

[0041]

[0042] <Chemical Formula 1-2>

[0043]

[0044] <Chemical Formula 1-3>

[0045]

[0046] <Chemical Formula 1-4>

[0047]

[0048] <Chemical Formula 1-5>

[0049]

[0050] <Chemical Formula 1-6>

[0051]

[0052] Among the above chemical formulas 1-1 to 1-6,

[0053] Z, R 11 Inland R 13 , R 21 Inland R 24 , R3 to R6 and Q1 are each the same as described above,

[0054] *is (CL1) in the above chemical formula I c1 -(SL1) s1 -(CL2) c2 -(SL2) s2 -This is a site that is combined with D.

[0055] According to one embodiment, D in the above chemical formula I is

[0056] Camptothecin, paclitaxel, doxorubicin, auristatin E, auristatin F, monomethyl auristatin E (MMAE), monomethyl auristatin F (MMAF), cyclophosphamide, mechlorethamine, uramustine, melphalan, chlorambucil, ifosfamide, bendamustine, carmustine, lomustine, streptozocin, busulfan, dacarbazine, temozolomide, thiotepa, altretamine, duocarmycin, cisplatin, carboplatin, nedaplatin, oxaliplatin, satraplatin, triplatin tetranitrate, 5-fluorouracil, 6-mercaptopurine, capecitabine, cladribine, clofarabine, cystabine, floxuridine, fludarabine, gemcitabine, hydroxyurea, methotrexate, pemetrexed, pentostatin, thioguanine, topotecan, exatecan,It may be at least one selected from the group consisting of irinotecan, etoposide, teniposide, mitoxantrone, doxetaxel, izabepilone, vinblastine, vincristine, vindesine, vinorelbine, estramustine, maytansine, mertansine, dolastatin, and derivatives thereof. For example, D may be selected from the group consisting of 7-ethyl-10-hydroxy camptothecin (SN38), which is a camptothecin derivative, exatecan, paclitaxel, doxorubicin, MMAE, which are camptothecin derivatives, and derivatives thereof.

[0057] According to one embodiment, the sum of c1, s1, c2 and s2 in the above formula I can be 0, 1, 2 or 3. The sum of c1 and c2 can be 0 or 1. The sum of s1 and s2 can be 0 or 1. For example, c1 can be 1, s1 can be 0, c2 can be 0 and s2 can be 0. For another example, c1 can be 0, s1 can be 1, c2 can be 0 and s2 can be 0. For yet another example, c1 can be 0, s1 can be 1, c2 can be 1 and s2 can be 0.

[0058] According to one embodiment, in the above formula I, CL1 and CL2 may independently include a carbonyl group (-C(=O)-), an ester group (-C(=O)O-), an amide group (-C(=O)NH-), a carbonate group (-OC(=O)O-), a disulfide group (-SS-), and any combination thereof. For example, CL1 and CL2 may be * 1 -(CH2) p -SS-(CH2) q -* 2 (p and q are each integers from 0 to 20, *1 and * 2 Each may include a group represented by a bonding site with a neighboring atom. For example, p and q may be integers from 0 to 10, and specifically 0, 1, 2, 3, 4, or 5.

[0059] According to one embodiment, in the above chemical formula I, SL1 and SL2 are a carbonyl group (-C(=O)-), an ester group (-C(=O)O-), an amide group (-C(=O)NH-), a carbonate group (-OC(=O)O-), * 3 -(CH2CH2O) r -* 4 (r is an integer from 1 to 20, * 3 and * 4 Each may include a group represented by a bonding site with a neighboring atom, and any combination thereof. For example, r may be an integer from 4 to 10, and specifically may be 4, 5, 6, 7, or 8.

[0060] According to one embodiment, the albumin-binding prodrug can bind to albumin present in the blood through injection administration to form an albumin-drug conjugate. However, the method of administering the albumin-binding prodrug may include not only injection administration, but also oral administration, topical administration, implantation administration, nasal administration, sublingual administration, inhalation administration, transdermal administration, intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, intranasal administration, intrapulmonary administration, and rectal administration. The albumin-binding prodrug can be administered to a subject and bind to albumin present in the subject. The subject includes any human or non-human animal. The non-human animal may be a vertebrate, such as a primate, sheep, dog, cow, or a rodent, such as a mouse, rat, or guinea pig.

[0061] According to one aspect, a pharmaceutical composition for preventing or treating cancer is disclosed, comprising the above-described albumin-binding prodrug as an active ingredient. The content of the active ingredient in the pharmaceutical composition of the present disclosure is not particularly limited, but may be 0.01 μM or more, for example, 0.1 μM or more, 0.5 μM or more, 1 μM or more, 2 μM or more, 5 μM or more, 10 μM or more, 20 μM or more, 50 μM or more, or 100 μM or more.

[0062] According to one embodiment, the cancer is selected from the group consisting of stomach cancer, lung cancer, non-small cell lung cancer, breast cancer, kidney cancer, ovarian cancer, liver cancer, bronchial cancer, nasopharyngeal cancer, oral cancer, laryngeal cancer, pancreatic cancer, bladder cancer, colon cancer, cervical cancer, bone cancer, non-small cell bone cancer, blood cancer, skin cancer, head or neck cancer, uterine cancer, rectal cancer, colon cancer, anal cancer, fallopian tube cancer, endometrial cancer, vaginal cancer, fallopian tube cancer, vulvar cancer, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, chronic or acute leukemia, lymphocytic lymphoma, kidney or ureter cancer, renal cell carcinoma, renal pelvic carcinoma, salivary gland cancer, sarcoma, pseudomyxoma, hepatoblastoma, testicular cancer, glioblastoma, lip cancer, It may be at least one selected from the group consisting of ovarian germ cell tumor, basal cell carcinoma, multiple myeloma, gallbladder cancer, choroidal melanoma, ampulla of Vater cancer, peritoneal cancer, tongue cancer, small cell carcinoma, pediatric lymphoma, neuroblastoma, duodenal cancer, ureteral cancer, astrocytoma, meningioma, renal pelvis cancer, vulvar cancer, thymic cancer, central nervous system tumor, primary central nervous system lymphoma, spinal cord tumor, brainstem glioma, and pituitary adenoma.

[0063] According to one aspect, a method for preparing an albumin-drug conjugate is disclosed, comprising a step of reacting an albumin-binding prodrug represented by the above-described chemical formula I with albumin. The albumin may be present within a subject or may be present outside the subject. When the albumin-binding prodrug is administered into a subject, it may react with albumin present within the subject to produce an albumin-drug conjugate. The albumin-binding prodrug may be reacted with albumin outside the subject to produce an albumin-drug conjugate, and the produced albumin-drug conjugate may be administered into the subject.

[0064] According to one aspect, an albumin-drug conjugate represented by the following chemical formula II is disclosed:

[0065] <Chemical Formula II>

[0066] Ab-(CL1) c1 -(SL1) s1 -(CL2) c2 -(SL2) s2 -D

[0067] In the above chemical formula II,

[0068] Ab is an albumin binding moiety represented by the following chemical formula 2,

[0069] CL1 and CL2 are each a cleavable linker,

[0070] SL1 and SL2 are each soluble linkers,

[0071] c1, c2, s1 and s2 are each 0 or 1,

[0072] D is a drug moiety,

[0073] <Chemical Formula 2>

[0074]

[0075] In the above chemical formula 2,

[0076] A is a group excluding amine groups in albumin,

[0077] Z is N, O or C(R 14 ) and,

[0078] R 11 Inland R 14 , R 21 Inland R 24 And R3 to R6 are each independently hydrogen, deuterium, -F, -Cl, -Br, -I, hydroxyl group, cyano group, nitro group, formyl group, carboxyl group, amino group, substituted or unsubstituted C1-C 10 Alkyl group, substituted or unsubstituted C2-C 10 Alkenyl group, substituted or unsubstituted C2-C 10 Alkynyl group, substituted or unsubstituted C1-C 10 Alkoxy group, substituted or unsubstituted C6-C 10 Aryl group, -N(Q1)(Q2) and -C(=O)(Q1), selected from

[0079] R 11 Inland R 14 and R 21 Inland R 24 One of them is (CL1) in the above chemical formula II c1 -(SL1) s1 -(CL2) c2 -(SL2) s2 -This is a site that combines with D,

[0080] Optionally i) R 11 and R 12 , ii) R 12 Wow R 13 , iii) R 13 and R 14 , iv) R 21 and R 22 , v) R 22 Wow R 23 , vi) R 23 and R 24 , or vii) any combination thereof, which may be combined with each other to form a substituted or unsubstituted C3-C 10 Carbocyclic group or substituted or unsubstituted C1-C 10 Forming a heterocyclic group,

[0081] Q1 and Q2 are independently hydrogen, deuterium, -F, -Cl, -Br, -I, hydroxyl group, cyano group, nitro group, formyl group, carboxyl group, amino group, substituted or unsubstituted C1-C 10 Alkyl group, substituted or unsubstituted C2-C 10 Alkenyl group, substituted or unsubstituted C2-C 10 Alkynyl group, substituted or unsubstituted C1-C 10 Alkoxy group and substituted or unsubstituted C6-C 10 Selected from aryl groups,

[0082] substituted C1-C 10 Alkyl group, substituted C2-C 10 Alkenyl group, substituted C2-C 10 alkynyl group, substituted C1-C 10 Alkoxy group, substituted C6-C 10 Aryl group, substituted C3-C 10 Carbocyclic groups and substituted C1-C 10 The substituents of the heterocyclic group are deuterium, -F, -Cl, -Br, -I, hydroxyl group, cyano group, nitro group, formyl group, carboxyl group, amino group, C1-C 10 Alkyl group and C6-C 10 Selected from aryl groups,

[0083] m is an integer selected from 1 to 5.

[0084] The albumin-binding moiety Ab represented by the above chemical formula 2 may be a moiety formed by binding the albumin-binding moiety Ac represented by the above chemical formula 1 to albumin. Therefore, in the above chemical formula 2, Z, R 11 Inland R 14 , R 21 Inland R 24 , the description for each of R3 to R6 and m is the same as that described in the chemical formula 1 described above.

[0085] In the above chemical formula 2, A is a group excluding an amine group in albumin, which may be located at the sudlow site I (SS1) site in albumin, and the amine group may be an amine group included in lysine 199 located at the SS1 site of albumin.

[0086] According to one aspect, a pharmaceutical composition for preventing or treating cancer is disclosed, comprising the above-described albumin-drug conjugate as an active ingredient. The content of the active ingredient is the same as described above.

[0087] The pharmaceutical composition for preventing or treating cancer, which comprises an albumin-binding prodrug represented by the above-described chemical formula I as an active ingredient, and the pharmaceutical composition for preventing or treating cancer, which comprises an albumin-drug conjugate represented by the above-described chemical formula II as an active ingredient, may independently include a solvent, an excipient, an additive, an injection, a buffer solution, etc., and may specifically be in the form of an aqueous solution.

[0088] Since albumin can be selectively accumulated in cancer tumors, prodrugs and albumin-drug conjugates that can stably and selectively bind to albumin and are stably bound to anticancer drugs at the same time can target cancer tumors and deliver anticancer drugs.

[0089] The above prodrug forms an albumin-drug conjugate by binding to albumin without the addition of any substance other than albumin, so an additional purification process is not necessary, making the manufacturing process easy and economical, and there are no separate restrictions on use, making it highly useful.

[0090] In addition, while general anticancer drugs have low water solubility, the above-described prodrugs and albumin-drug conjugates have improved water solubility and can be taken in the form of an aqueous solution, making them advantageous for use as injections, etc.

[0091] Figure 1 is a schematic diagram illustrating the operating principle of the albumin-binding prodrug and albumin-drug conjugate of the present disclosure.

[0092] Figure 2 is a schematic diagram showing the structure in which FLIC and albumin of Figure 1 bind at the SS1 site.

[0093] Figure 3a is a diagram showing changes in the UV-Vis spectrum of compound 19 (P-PEG8) depending on the presence or absence of albumin.

[0094] Figure 3b is a diagram showing changes in the fluorescence spectrum according to changes in the concentration of albumin mixed with compound 19 (P-PEG8).

[0095] Figure 3c is a diagram showing the change in the UV-Vis spectrum of compound 9 (P-SS-COOH) depending on the presence or absence of albumin.

[0096] Figure 3d is a diagram showing changes in the fluorescence spectrum according to changes in the concentration of albumin mixed with compound 9 (P-SS-COOH).

[0097] Figure 4a is a diagram showing the change in the UV-Vis spectrum of compound 12 (P-SS-SN38) depending on the presence or absence of albumin.

[0098] Figure 4b is a diagram showing changes in the fluorescence spectrum according to changes in the concentration of albumin mixed with compound 12 (P-SS-SN38).

[0099] Figure 5a is a diagram showing changes in the UV-Vis spectrum of compound 15 (P-SS-Pac) depending on the presence or absence of albumin.

[0100] Figure 5b is a diagram showing changes in the fluorescence spectrum of compound 15 (P-SS-Pac) depending on the presence or absence of albumin.

[0101] Figure 6a is a diagram showing changes in UV-Vis spectra according to changes in the concentration of albumin mixed with compound 18 (P-SS-Doxo).

[0102] Figure 6b is a diagram showing the change in fluorescence spectrum over time according to the change in the concentration of albumin mixed with compound 18 (P-SS-Doxo).

[0103] Figure 7a is a diagram showing changes in the UV-Vis spectrum of compound 26A (P-PEG8-SN38) depending on the presence or absence of albumin.

[0104] Figure 7b is a diagram showing changes in the fluorescence spectrum according to changes in the concentration of albumin mixed with compound 26A (P-PEG8-SN38).

[0105] Figure 8a is a diagram showing changes in the UV-Vis spectrum of compound 26C (P-PEG8-Doxo) depending on the presence or absence of albumin.

[0106] Figure 8b is a diagram showing changes in the fluorescence spectrum according to the concentration of albumin mixed with compound 26C (P-PEG8-Doxo).

[0107] Figure 9a is a diagram showing changes in the UV-Vis spectrum of compound 26B (P-PEG8-Pac) depending on the presence or absence of albumin.

[0108] Figure 9b is a diagram showing changes in the fluorescence spectrum according to changes in the concentration of albumin mixed with compound 26B (P-PEG8-Pac).

[0109] Figure 10a is a diagram showing changes in the UV-Vis spectrum of compound 26D (P-PEG8-MMAE) depending on the presence or absence of albumin.

[0110] Figure 10b is a diagram showing changes in the fluorescence spectrum according to changes in the concentration of albumin mixed with compound 26D (P-PEG8-MMAE).

[0111] Figure 11a is a diagram showing changes in the UV-Vis spectrum of compound 23 (P-PEG8-SS-SN38) depending on the presence or absence of albumin.

[0112] Figure 11b is a diagram showing changes in the fluorescence spectrum according to changes in the concentration of albumin mixed with compound 23 (P-PEG8-SS-SN38).

[0113] Figure 12a is a diagram showing changes in the UV-Vis spectrum of compound 31A (P-3C-SS-SN38) depending on the presence or absence of albumin.

[0114] Figure 12b is a diagram showing changes in the fluorescence spectrum according to changes in the concentration of albumin mixed with compound 31A (P-3C-SS-SN38).

[0115] Figure 13a is a diagram showing changes in the UV-Vis spectrum of compound 31D (P-3C-SS-MMAE) depending on the presence or absence of albumin.

[0116] Figure 13b is a diagram showing changes in the fluorescence spectrum according to changes in the concentration of albumin mixed with compound 31D (P-3C-SS-MMAE).

[0117] Figure 14a is a diagram showing changes in the UV-Vis spectrum of compound 31B (P-3C-SS-Pac) depending on the presence or absence of albumin.

[0118] Figure 14b is a diagram showing changes in the fluorescence spectrum according to changes in the concentration of albumin mixed with compound 31B (P-3C-SS-Pac).

[0119] Figures 15a to 15d are drawings showing the results of in vitro anticancer activity evaluations performed on HeLA cell lines and MDA MB 231 cell lines, respectively, by applying a compound without an anticancer agent bound to it, an anticancer agent, an albumin-bound prodrug with an anticancer agent bound to it, and an albumin-drug conjugate with an anticancer agent bound to it.

[0120] Figures 16a to 16d are drawings showing the results of in vitro anticancer activity evaluation using FLIC-drug conjugates in which each anticancer agent is bound to a single FLIC for HeLA cell lines.

[0121] Figures 17a to 17d are drawings showing the results of in vitro anticancer activity evaluation using FLIC-drug conjugates in which each anticancer drug is bound to a single FLIC against the MDA MB 231 cell line.

[0122] Figures 18a to 18d are drawings showing the results of in vitro anticancer activity evaluation using FLIC-drug conjugates in which each anticancer drug is bound to a different FLIC for HeLA cell lines.

[0123] Figures 19a to 19d are drawings showing the results of in vitro anticancer activity evaluation using FLIC-drug conjugates in which each anticancer drug is bound to a different FLIC for the MDA MB 231 cell line.

[0124] Figure 20 is a drawing showing the results of an in vitro anticancer activity evaluation using a FLIC-drug conjugate in which each anticancer drug is bound to FLIC for the HCT116 cell line.

[0125] Figure 21 is a schematic diagram of the in vivo anticancer activity evaluation for mice transplanted with the MDA MB 231 cell line.

[0126] Figures 22a to 22d are drawings showing the results of the in vivo anticancer activity evaluation of Figure 21.

[0127] Unless otherwise defined, all technical terms used in this specification have the same meaning as commonly understood by those skilled in the art. Furthermore, numerical values ​​described herein are assumed to include the meaning of "about," even if not explicitly stated.

[0128] The term "include" in this specification is used to indicate that other components may be added and / or interposed, rather than to the exclusion of other components, unless specifically stated otherwise.

[0129] As used herein, the term “any combination” means a mixture or combination of one or more of the described components.

[0130] In this specification, "C a -C b " means that a particular functional group can contain from a to b number of carbon atoms.

[0131] As used herein, "alkyl group" means a branched or unbranched aliphatic hydrocarbon. The alkyl group may include a methyl group, an ethyl group, a propyl group, an isopropyl group, a butyl group, an isobutyl group, and the like.

[0132] In this specification, an "alkenyl group" means a monovalent hydrocarbon group having at least one carbon-to-carbon double bond in an alkyl group having two or more carbon atoms among the above "alkyl groups." The alkenyl group may include a vinyl group, an allyl group, an isopropenyl group, a butenyl group, and the like.

[0133] In this specification, “alkynyl group” means a monovalent hydrocarbon group having at least one carbon-to-carbon triple bond in an alkyl group having two or more carbon atoms among the above “alkyl groups.”

[0134] In this specification, "alkoxy group" refers to a form in which the above "alkyl group" is bonded to an oxygen atom. The alkoxy group may include a methoxy group, an ethoxy group, a propoxy group, a butoxy group, etc.

[0135] As used herein, "aryl group" refers to a monovalent functional group formed by the removal of a hydrogen atom present in an arene ring. The aryl group may include a phenyl group, a biphenyl group, a terphenyl group, a naphthyl group, and the like.

[0136] The term “albumin” as used herein may be bovine serum albumin (BSA), mouse serum albumin (MSA), or human serum albumin (HSA).

[0137] The pharmaceutical composition of the present invention can be administered in a pharmaceutically effective amount. The pharmaceutically effective amount includes a "therapeutically effective amount" and a "prophylactically effective amount." The term "therapeutically effective amount" refers to any amount of a drug or therapeutic agent that, when used alone or in combination with other therapeutic agents, can exhibit a decrease in the severity of disease symptoms, an increase in the frequency and duration of disease symptom-free periods, or a prevention of impairment or disability due to disease affliction. The term "prophylactically effective amount" refers to any amount of a drug that inhibits the development, metastasis, or recurrence of cancer in a subject at risk of developing cancer or suffering from cancer metastasis or recurrence. The level of the effective amount can be determined based on factors such as the subject's type and severity, age, sex, activity of the drug, sensitivity to the drug, time of administration, route of administration, and excretion rate, duration of treatment, concurrently used drugs, and other factors well known in the medical field.

[0138] In this specification, “albumin-binding prodrug” and “albumin-drug conjugate” are substances to which a drug, such as an anticancer agent, is directly bound and can prevent or treat cancer through the drug, and are therefore clearly different from “imaging cancer,” “treating cancer by generating reactive oxygen species,” or “photodynamic therapy based on the premise of light irradiation.”

[0139] According to one aspect, an albumin-binding prodrug represented by the following chemical formula I is disclosed:

[0140] <Chemical Formula I>

[0141] Ac-(CL1) c1 -(SL1) s1 -(CL2) c2 -(SL2) s2 -D

[0142] In the above chemical formula I,

[0143] Ac is an albumin-binding moiety represented by the following chemical formula 1,

[0144] CL1 and CL2 are each a cleavable linker,

[0145] SL1 and SL2 are each soluble linkers,

[0146] c1, c2, s1 and s2 are each 0 or 1,

[0147] D is a drug moiety,

[0148] <Chemical Formula 1>

[0149]

[0150] In the above chemical formula 1,

[0151] Z is N, O or C(R 14 ) and,

[0152] R 11 Inland R 14 , R 21 Inland R 24 And R3 to R6 are each independently hydrogen, deuterium, -F, -Cl, -Br, -I, hydroxyl group, cyano group, nitro group, formyl group, carboxyl group, amino group, substituted or unsubstituted C1-C 10 Alkyl group, substituted or unsubstituted C2-C 10 Alkenyl group, substituted or unsubstituted C2-C 10 Alkynyl group, substituted or unsubstituted C1-C 10 Alkoxy group, substituted or unsubstituted C6-C 10 Aryl group, -N(Q1)(Q2) and -C(=O)(Q1), selected from

[0153] R 11 Inland R 14 and R 21 Inland R 24 One of them is (CL1) in the above chemical formula I c1 -(SL1) s1 -(CL2) c2 -(SL2) s2 -This is a site that combines with D,

[0154] Optionally i) R 11and R 12 , ii) R 12 Wow R 13 , iii) R 13 and R 14 , iv) R 21 and R 22 , v) R 22 Wow R 23 , vi) R 23 and R 24 , or vii) any combination thereof, which may be combined with each other to form a substituted or unsubstituted C3-C 10 Carbocyclic group or substituted or unsubstituted C1-C 10 Forming a heterocyclic group,

[0155] Q1 and Q2 are independently hydrogen, deuterium, -F, -Cl, -Br, -I, hydroxyl group, cyano group, nitro group, formyl group, carboxyl group, amino group, substituted or unsubstituted C1-C 10 Alkyl group, substituted or unsubstituted C2-C 10 Alkenyl group, substituted or unsubstituted C2-C 10 Alkynyl group, substituted or unsubstituted C1-C 10 Alkoxy group and substituted or unsubstituted C6-C 10 Selected from aryl groups,

[0156] substituted C1-C 10 Alkyl group, substituted C2-C 10 Alkenyl group, substituted C2-C 10 alkynyl group, substituted C1-C 10 Alkoxy group, substituted C6-C 10 Aryl group, substituted C3-C 10 Carbocyclic groups and substituted C1-C 10 The substituents of the heterocyclic group are deuterium, -F, -Cl, -Br, -I, hydroxyl group, cyano group, nitro group, formyl group, carboxyl group, amino group, C1-C 10 Alkyl group and C6-C 10 Selected from aryl groups,

[0157] m is an integer selected from 1 to 5.

[0158] Among the above chemical formulas (I), the albumin-binding moiety Ac can selectively react with albumin and bind to albumin. That is, the albumin-binding prodrug represented by the above chemical formula (I) can bind to albumin when mixed with albumin in an aqueous solution to form an albumin-drug conjugate.

[0159] The albumin-binding moiety Ac represented by the above chemical formula 1 can selectively react with albumin and bind to albumin. In this case, the albumin-binding moiety Ac can bind to albumin to become the albumin-binding moiety Ab represented by the above chemical formula 2, and the albumin-binding prodrug represented by the above chemical formula I can become an albumin-drug conjugate represented by the above chemical formula II.

[0160] As well as the albumin-binding moiety Ac before binding to albumin and the albumin-binding moiety Ab after binding to albumin, they are (CL1) in the above formulae I and II. c1 -(SL1) s1 -(CL2) c2 -(SL2) s2 -Compounds in which the bond is broken with the group indicated by -D can be collectively referred to as FLIC. That is, the albumin-binding prodrug indicated by the above chemical formula I and the albumin-drug conjugate indicated by the above chemical formula II can both be referred to as FLIC-drug conjugates.

[0161] An albumin-binding moiety Ac represented by the above chemical formula 1 or (CL1) of the above chemical formula I within a specific compound c1 -(SL1) s1 -(CL2) c2 -(SL2) s2 When the albumin-binding moiety Ac exists in the form of a compound in which the bond is broken with the group indicated by -D, it can be indicated by writing "P-" in the name of the compound.

[0162] An albumin binding moiety Ab represented by the above chemical formula 2 or (CL1) of the above chemical formula II within a specific compound; c1 -(SL1) s1 -(CL2) c2 -(SL2) s2 -If there is an albumin-binding moiety Ab in the form of a compound in which the bond is broken with the group indicated by -D, the name of the compound may be indicated by indicating the type of albumin, in addition to "P-", by indicating "HSA", "BSA" or "MSA", in addition to "P-", since more albumin is bound.

[0163] Figure 1 is a schematic diagram illustrating the operating principle of the albumin-binding prodrug and albumin-drug conjugate of the present disclosure.

[0164] Referring to Fig. 1, the FLIC-drug conjugate can bind to the amine group present in lysine 199 (Lys199) present in the sudlow site I (SS1) of albumin. Due to the structural specificity of FLIC and the positional specificity of lysine 199 in the SS1 site in albumin, the FLIC-drug conjugate can bind relatively strongly to albumin. The albumin-drug conjugate bound to albumin through FLIC can accumulate in cancer due to the cancer-targeting property of albumin. The albumin-drug conjugate accumulated in the cancer can release the drug and treat the cancer. Therefore, the FLIC-drug conjugate, which collectively refers to the albumin-binding prodrug represented by the above chemical formula I and the albumin-drug conjugate represented by the above chemical formula II, can effectively prevent or treat cancer, and can prevent the drug-induced toxicity from increasing in normal cells where cancer cells do not exist.

[0165] Figure 2 is a schematic diagram showing the structure in which FLIC and albumin of Figure 1 bind at the SS1 site.

[0166] Referring to FIG. 2, the albumin-binding moiety Ac represented by the chemical formula 1 may bind to the amine group of lysine 199 located at the SS1 site of albumin to become the albumin-binding moiety Ab represented by the chemical formula 2. In this case, based on the albumin-binding moiety Ac represented by the chemical formula 1, the benzene group to which boron is directly bonded may face the entrance direction of the SS1 site of albumin, and the 6-membered ring to which the carbonyl group is directly bonded may face the inner direction of the SS1 site of albumin. Therefore, when a drug is linked to the benzene group to which boron is directly bonded in FLIC, FLIC and albumin can be easily bonded without structural interference. That is, in the chemical formulas 1 and 2, R 21 Inland R 24 (CL1) in any one of the above chemical formulas I and II c1 -(SL1) s1 -(CL2) c2 -(SL2) s2 -The group marked with -D may be structurally stable when combined.

[0167] FLIC (CL1) c1 -(SL1) s1 -(CL2) c2 -(SL2) s2 Before being linked to a group indicated by -D, FLIC may contain a functional group such as -COOH, -OH, -NH2 at the terminal to link to such a group. For example, FLIC may have a structure selected from, but not limited to, Group 1 below:

[0168] Group 1

[0169] .

[0170] As a non-limiting example, the FLIC selected from Group 1 above may be linked to a drug, wherein the drug may be an anticancer agent selected from Group 2 below, but is not limited thereto:

[0171] Group 2

[0172] .

[0173] SN38 is a camptothecin derivative, which may be 7-ethyl-10-hydroxy camptothecin. When the FLIC-drug conjugate comprises SN38, the compound name may be designated to include "SN38." When the FLIC-drug conjugate comprises paclitaxel, the compound name may be designated to include "Pac." When the FLIC-drug conjugate comprises doxorubicin, the compound name may be designated to include "Doxo." When the FLIC-drug conjugate comprises monomethyl auristatin E, the compound name may be designated to include "MMAE."

[0174] FLIC and the drug can be directly linked to each other. Alternatively, a linker can be introduced between the FLIC and the drug to improve the solubility of the FLIC-drug conjugate, enhance binding due to the spatial structure of the SS1 position within albumin, or improve drug release. The linker can include a cleavable linker (CL), a soluble linker (SL), or any combination thereof.

[0175] The above-mentioned cleavable linker (CL) can play a role in releasing anticancer drugs from cancer cells, thereby exerting anticancer therapeutic effects. For example, the cleavable linker (CL) can be degraded by glutathione (GSH), which is known to be present in cancerous tumors. Since the pH of cancerous tumors is relatively low, the cleavable linker (CL) may include a functional group that has the property of being degraded at relatively low pH. The cleavable linker (CL) can directly bind to the anticancer drug and may have the property of being cleaved in cancerous cells but not cleaved in normal cells. The cleavable linker (CL) may not be present, and a noncleavable linker may be applied depending on the type of anticancer drug. The cleavable linker (CL) may include a carbonyl group (-C(=O)-), an ester group (-C(=O)O-), an amide group (-C(=O)NH-), a carbonate group (-OC(=O)O-), a disulfide group (-SS-), and any combination thereof. The cleavable linker (CL) may be * 1 -(CH2) p -SS-(CH2) q -* 2 (p and q are each integers from 0 to 20, * 1 and * 2 Each may contain a group indicated as a binding site with a neighboring atom. FLIC-drug conjugates are * 1 -(CH2) p -SS-(CH2) q -* 2 When a cleavable linker (CL) is included, the compound name may be referred to by including "SS". For example, a cleavable linker (CL) may be indicated by * 1 -(CH2) p -SS-(CH2) q -* 2 It may include a group indicated by , or may further include functional groups such as a carbonyl group, an ester group, an amide group, or a carbonate group at both ends thereof.

[0176] The above soluble linker (SL) can play a role in increasing the water solubility of the FLIC-drug conjugate. The FLIC-drug conjugate with increased water solubility can have improved binding to albumin. The soluble linker (SL) may or may not be present. The soluble linker (SL) may have the same or similar structure as polyethylene glycol (PEG). For example, the soluble linker (SL) may be * 3 -(CH2CH2O) r -* 4 (r is an integer from 1 to 20, * 3 and * 4 Each may contain a group indicated as a bonding site with a neighboring atom. The soluble linker (SL) is * 3 -(CH2CH2O) r -* 4 When a group is indicated by r, it can be specifically referred to according to the value of r. For example, if r is 4, the soluble linker (SL) can be referred to as "PEG4". For another example, if r is 8, the soluble linker (SL) can be referred to as "PEG8". The soluble linker (SL) is * 3 -(CH2CH2O) r -* 4 It may include a group indicated by , or may further include functional groups such as a carbonyl group, an ester group, an amide group, or a carbonate group at both ends thereof.

[0177] The cleavable linker (CL) and the soluble linker (SL) may independently include a functional group for bonding with a terminally adjacent group. For example, the functional group may be an ester group, an amide group, a carbonate group, a carbonyl group, a click group, etc. The click group may include a group capable of undergoing a click reaction (e.g., an azide group and an alkyne group) or a group formed by a click reaction (e.g., a triazole group).

[0178] The albumin-binding prodrug represented by the above chemical formula I can form an albumin-drug conjugate represented by the above chemical formula II when mixed with albumin in an aqueous solution. In such a composition, the pH of the aqueous solution may be 6 to 9. The aqueous solution may be a buffer solution having a pH of 7.4, similar to the pH of blood. If the pH of the aqueous solution is less than 6, the binding between FLIC and albumin may be relatively weak, preventing the formation of the conjugate. If the pH of the aqueous solution exceeds 9, albumin may be denatured, thereby reducing the cancer-targeting function.

[0179] The boron of the albumin-binding moiety Ac represented by the above chemical formula 1 is not protected with pinacol. The albumin-binding moiety Ac undergoes the following reversible reaction in the presence of albumin in an aqueous solution, and the boron is protected with pinacol to form pinacolborane:

[0180]

[0181] Both non-pinacol-protected and pinacol-protected structures can fall under the term FLIC, and can be referred to as FLIC-deprotected and FLIC-protected, respectively. Both FLIC-protected and FLIC-deprotected structures exist in aqueous solution due to a reversible reaction, and while FLIC-protected structures may not bind to albumin, FLIC-deprotected structures can bind to albumin.

[0182] The binding of albumin to FLIC is reversible and reaches equilibrium, and the binding strength is given by the binding constant (K B ) or dissociation constant (K D ) can be expressed as.

[0183] The albumin-binding moiety Ac represented by the above chemical formula 1 may not exhibit fluorescence. The albumin-binding moiety Ab represented by the above chemical formula 2 may exhibit fluorescence. That is, whether it binds to albumin can be determined by changes in the UV-Vis signal and the fluorescence signal. By using these characteristics, when the fluorescence signal or the UV-Vis signal is analyzed according to changes in the concentration of albumin, the dissociation constant (K D ) can measure the value.

[0184] [Example]

[0185] Figure 3a is a diagram showing changes in the UV-Vis spectrum of compound 19 (P-PEG8) depending on the presence or absence of albumin. Figure 3b is a diagram showing changes in the fluorescence spectrum depending on changes in the concentration of albumin mixed with compound 19 (P-PEG8). Compound 19 (P-PEG8) is a compound in which PEG8, a soluble linker, is bound to FLIC, and can be confirmed in Scheme 5 described below.

[0186] Referring to Figures 3a and 3b, compound 19 (P-PEG8) reacts with albumin (HSA) and causes significant changes in UV-Vis signals and fluorescence signals, indicating that it effectively binds to albumin. Based on the change in fluorescence signal according to the change in albumin concentration, the dissociation constant (K) of compound 19 (P-PEG8) and albumin D ) was measured to be 4.5 μM. Therefore, it can be seen that an albumin-drug conjugate can be prepared by linking an anticancer drug to compound 19 (P-PEG8).

[0187] Figure 3c is a diagram showing changes in the UV-Vis spectrum of compound 9 (P-SS-COOH) depending on the presence or absence of albumin. Figure 3d is a diagram showing changes in the fluorescence spectrum depending on changes in the concentration of albumin mixed with compound 9 (P-SS-COOH). Compound 9 (P-SS-COOH) is a compound in which a cleavable linker (SS) is bonded to FLIC and a COOH substituent is present at the terminal, and can be confirmed in Scheme 4 described below.

[0188] Referring to FIGS. 3c and 3d, compound 9 (P-SS-COOH) reacts with albumin and causes significant changes in UV-Vis signals and fluorescence signals, indicating that it is a compound that can effectively bind to albumin.

[0189] Figure 4a is a diagram showing changes in the UV-Vis spectrum of compound 12 (P-SS-SN38) depending on the presence or absence of albumin. Figure 4b is a diagram showing changes in the fluorescence spectrum depending on changes in the concentration of albumin mixed with compound 12 (P-SS-SN38). Compound 12 (P-SS-SN38) is a compound in which a cleavable linker (SS) is bound to FLIC and SN38 as an anticancer agent is connected to the terminal, and can be confirmed in Synthetic Example A1 described below.

[0190] Referring to Figures 4a and 4b, changes in UV-Vis signals and fluorescence signals are observed, but the degree of change is relatively small, and the solubility of compound 12 is relatively low. It was not possible to distinguish whether the change in signal occurred because the solubility of compound 12 increased with the addition of albumin or because a complex was formed with albumin.

[0191] Figure 5a is a diagram showing changes in the UV-Vis spectrum of compound 15 (P-SS-Pac) depending on the presence or absence of albumin. Figure 5b is a diagram showing changes in the fluorescence spectrum of compound 15 (P-SS-Pac) depending on the presence or absence of albumin. Compound 15 (P-SS-Pac) is a compound in which a cleavable linker (SS) is attached to FLIC and paclitaxel (Pac) as an anticancer agent is connected to the terminal, and can be confirmed in Synthetic Example A2 described below.

[0192] Referring to Figures 5a and 5b, changes in UV-Vis signals and fluorescence signals are observed, but the degree of change is relatively small, and the solubility of compound 15 is relatively low. It was not possible to distinguish whether the change in signal occurred because the solubility of compound 15 increased with the addition of albumin or because a complex with albumin was formed.

[0193] Figure 6a is a diagram showing changes in the UV-Vis spectrum according to changes in the concentration of albumin mixed with compound 18 (P-SS-Doxo). Figure 6b is a diagram showing changes in the fluorescence spectrum according to changes in the concentration of albumin mixed with compound 18 (P-SS-Doxo) over time. Compound 18 (P-SS-Doxo) is a compound in which a cleavable linker (SS) is attached to FLIC and an anticancer agent, doxorubicin (Doxo), is connected to the terminal, and can be confirmed in Synthetic Example A3 described below.

[0194] Referring to Figures 6a and 6b, changes in UV-Vis signals and fluorescence signals are observed, but the degree of change is relatively small, and the solubility of compound 18 is relatively low. It was not possible to distinguish whether the change in signal occurred as the solubility of compound 18 increased with the addition of albumin or because a complex was formed with albumin.

[0195] Figure 7a is a diagram showing changes in the UV-Vis spectrum of compound 26A (P-PEG8-SN38) depending on the presence or absence of albumin. Figure 7b is a diagram showing changes in the fluorescence spectrum depending on changes in the concentration of albumin mixed with compound 26A (P-PEG8-SN38). Compound 26A (P-PEG8-SN38) is a compound in which a soluble linker (PEG8) is bound to FLIC and SN38 as an anticancer agent is connected to the terminal, and can be confirmed in Synthetic Example C1 described below.

[0196] Referring to FIGS. 7a and 7b, compound 26A can be easily observed to have a UV-Vis signal even without albumin, which may be due to its relatively high water solubility, and since the fluorescence signal changes significantly upon addition of albumin, it can be seen that it has excellent binding affinity to albumin.

[0197] Figure 8a is a diagram showing the change in the UV-Vis spectrum of compound 26C (P-PEG8-Doxo) depending on the presence or absence of albumin. Figure 8b is a diagram showing the change in the fluorescence spectrum depending on the concentration of albumin mixed with compound 26C (P-PEG8-Doxo). Compound 26C (P-PEG8-Doxo) is a compound in which a soluble linker (PEG8) is bound to FLIC and an anticancer agent, doxorubicin (Doxo), is connected to the terminal, and can be confirmed in Synthesis Example C3 described below.

[0198] Referring to Figures 8a and 8b, compound 26C can be easily observed with UV-Vis signals even without albumin, which may be due to its relatively high water solubility, and since the fluorescence signal changes significantly upon addition of albumin, it can be seen that it has excellent binding affinity with albumin. Based on the change in the fluorescence signal in Figure 8b, the binding dissociation constant is calculated as K D The value was approximately 3.6 μM.

[0199] Figure 9a is a diagram showing changes in the UV-Vis spectrum of compound 26B (P-PEG8-Pac) depending on the presence or absence of albumin. Figure 9b is a diagram showing changes in the fluorescence spectrum depending on changes in the concentration of albumin mixed with compound 26B (P-PEG8-Pac). Compound 26B (P-PEG8-Pac) is a compound in which a soluble linker (PEG8) is bound to FLIC and paclitaxel (Pac) as an anticancer agent is connected to the terminal, and can be confirmed in Synthesis Example C2 described below.

[0200] Referring to Figures 9a and 9b, compound 26B can be easily observed with UV-Vis signals even without albumin, which may be due to its relatively high water solubility, and since the fluorescence signal changes significantly upon addition of albumin, it can be seen that it has excellent binding affinity with albumin. Based on the change in the fluorescence signal in Figure 9b, the binding-dissociation constant is calculated as K D The value was approximately 1.1 μM.

[0201] Figure 10a is a diagram showing changes in the UV-Vis spectrum of compound 26D (P-PEG8-MMAE) depending on the presence or absence of albumin. Figure 10b is a diagram showing changes in the fluorescence spectrum depending on changes in the concentration of albumin mixed with compound 26D (P-PEG8-MMAE). Compound 26D (P-PEG8-MMAE) is a compound in which a soluble linker (PEG8) is bound to FLIC and monomethyl auristatin E (MMAE) as an anticancer agent is connected to the terminal.

[0202] Referring to Figures 10a and 10b, compound 26D can be easily observed with UV-Vis signals even without albumin, which may be due to its relatively high water solubility, and since the fluorescence signal changes significantly upon addition of albumin, it can be seen that it has excellent binding affinity with albumin. Based on the change in the fluorescence signal in Figure 10b, the binding dissociation constant is calculated as K D The value was approximately 2.4 μM.

[0203] Figure 11a is a diagram showing changes in the UV-Vis spectrum of compound 23 (P-PEG8-SS-SN38) depending on the presence or absence of albumin. Figure 11b is a diagram showing changes in the fluorescence spectrum depending on changes in the concentration of albumin mixed with compound 23 (P-PEG8-SS-SN38). Compound 23 (P-PEG8-SS-SN38) is a compound in which a soluble linker (PEG8) is bound to FLIC, followed by a cleavable linker (SS), and SN38 as an anticancer agent is connected to the terminal, and can be confirmed in Synthetic Example B1 described below.

[0204] Referring to Figures 11a and 11b, compound 23 has a very weak fluorescence signal without albumin, but when albumin is added, it shows very strong UV-Vis and fluorescence, indicating that it has excellent binding affinity with albumin. Based on the change in fluorescence signal in Figure 11b, the binding dissociation constant is calculated as K D The value was approximately 2 μM.

[0205] On the other hand, compound 24 (P-PEG8-SS-Pac) showed a relatively weak fluorescence change even upon addition of albumin.

[0206] In the case of paclitaxel (Pac), compound 15 bound to P-SS (see Figures 5a and 5b) has poor solubility and unclear binding to albumin, but compound 26B bound directly to P-PEG8 (see Figures 9a and 9b) has improved solubility and clearly exhibits binding to albumin.

[0207] Figure 12a is a diagram showing the change in the UV-Vis spectrum of compound 31A (P-3C-SS-SN38) depending on the presence or absence of albumin. Figure 12b is a diagram showing the change in the fluorescence spectrum depending on the change in the concentration of albumin mixed with compound 31A (P-3C-SS-SN38). Compound 31A (P-3C-SS-SN38) is a compound in which a linkage (3C) exists by a click reaction in FLIC, a cleavable linker (SS) is then attached, and SN38 as an anticancer agent is connected to the terminal, and can be confirmed in General Synthesis Example D described below.

[0208] Referring to Figures 12a and 12b, compound 31A has a very weak fluorescence signal without albumin, but when albumin is added, it shows very strong UV-Vis and fluorescence, indicating that it has excellent binding affinity with albumin. Based on the change in fluorescence signal in Figure 12b, the binding dissociation constant is calculated as K D The value was approximately 0.13 μM.

[0209] Figure 13a is a diagram showing the change in the UV-Vis spectrum of compound 31D (P-3C-SS-MMAE) depending on the presence or absence of albumin. Figure 13b is a diagram showing the change in the fluorescence spectrum depending on the change in the concentration of albumin mixed with compound 31D (P-3C-SS-MMAE). Compound 31D (P-3C-SS-MMAE) is a compound in which a linkage (3C) by a click reaction exists in FLIC, followed by a cleavable linker (SS) and MMAE as an anticancer agent is connected to the terminal, and can be confirmed in General Synthesis Example D described below.

[0210] Referring to Figures 13a and 13b, compound 31A has a very weak fluorescence signal without albumin, but when albumin is added, it shows very strong UV-Vis and fluorescence, indicating that it has excellent binding affinity with albumin.

[0211] Figure 14a is a diagram showing changes in the UV-Vis spectrum of compound 31B (P-3C-SS-Pac) depending on the presence or absence of albumin. Figure 14b is a diagram showing changes in the fluorescence spectrum depending on changes in the concentration of albumin mixed with compound 31B (P-3C-SS-Pac). Compound 31B (P-3C-SS-Pac) is a compound in which a linkage (3C) exists by a click reaction in FLIC, a cleavable linker (SS) is then attached, and paclitaxel (Pac) as an anticancer agent is connected to the terminal, and can be confirmed in General Synthesis Example D described below.

[0212] Referring to Figures 14a and 14b, compound 31A has a very weak fluorescence signal without albumin, but when albumin is added, it shows very strong UV-Vis and fluorescence, indicating that it has excellent binding affinity with albumin.

[0213] [in vitro anticancer activity evaluation]

[0214] The in vitro anticancer effects of various compounds synthesized in the synthetic examples described below were evaluated by expressing cell viability as a function of compound concentration. The evaluation was conducted using HeLa cell lines (human cervical cancer), MDA MB 231 cell lines (human triple-negative breast cancer), and HCT116 cell lines (human colon cancer).

[0215] Evaluation Example 1

[0216] Figures 15a to 15d are drawings showing the results of in vitro anticancer activity evaluations performed on HeLA cell lines and MDA MB 231 cell lines, respectively, by applying compound 9 (P-SS-COOH) to which an anticancer agent is not bound, anticancer agent SN38, compound 12 (P-SS-SN38) in the form of an albumin-bound prodrug linked to SN38, compound P-SS-SN38-HSA in the form of an albumin-drug conjugate linked to SN38, anticancer agent paclitaxel (Pac), compound 15 (P-SS-Pac) in the form of an albumin-bound prodrug linked to Pac, and compound P-SS-Pac-HSA in the form of an albumin-drug conjugate linked to Pac.

[0217] Referring to Fig. 15a, it can be confirmed that the FLIC-drug conjugate compounds P-SS-SN38 and P-SS-SN38-HSA exhibit excellent anticancer activity against the HeLA cell line.

[0218] Referring to Figure 15b, it can be confirmed that the FLIC-drug conjugate compounds P-SS-Pac and P-SS-Pac-HSA exhibit excellent anticancer activity against the HeLA cell line.

[0219] Referring to Fig. 15c, it can be confirmed that the FLIC-drug conjugates P-SS-SN38 and P-SS-SN38-HSA exhibit excellent anticancer activity against the MDA MB 231 cell line.

[0220] Referring to Figure 15d, it can be confirmed that the FLIC-drug conjugates P-SS-Pac and P-SS-Pac-HSA exhibit excellent anticancer activity against the MDA MB 231 cell line.

[0221] Evaluation Example 2

[0222] Figures 16a to 16d are diagrams showing the results of in vitro anticancer activity evaluations using FLIC-drug conjugates in which each anticancer drug (doxorubicin, MMAE, paclitaxel, SN38) is bound to FLIC for HeLA cell lines. Compound 19 (P-PEG8) with a soluble linker was used. That is, compound 26A (P-PEG8-SN38), compound 26B (P-PEG8-Pac), compound 26C (P-PEG8-Doxo), and compound 26D (P-PEG8-MMAE) were used.

[0223] Referring to Fig. 16a, it can be confirmed that the compound P-PEG8-Doxo in the form of an albumin-binding prodrug and the compound P-PEG8-Doxo-HSA in the form of an albumin-drug conjugate exhibit excellent anticancer activity against the HeLA cell line.

[0224] Referring to Figure 16b, it can be confirmed that the compound P-PEG8-MMAE in the form of an albumin-binding prodrug and the compound P-PEG8-MMAE-HSA in the form of an albumin-drug conjugate exhibit excellent anticancer activity against the HeLA cell line.

[0225] Referring to FIG. 16c, it can be confirmed that the compound P-PEG8-Pac in the form of an albumin-binding prodrug and the compound P-PEG8-Pac-HSA in the form of an albumin-drug conjugate exhibit excellent anticancer activity against the HeLA cell line.

[0226] Referring to FIG. 16d, it can be confirmed that the compound P-PEG8-SN38 in the form of an albumin-binding prodrug and the compound P-PEG8-SN38-HSA in the form of an albumin-drug conjugate exhibit excellent anticancer activity against the HeLA cell line.

[0227] Evaluation Example 3

[0228] Figures 17a to 17d are drawings showing the results of repeating the evaluation example 2 above by changing the HeLA cell line to the MDA MB 231 cell line.

[0229] Referring to Figure 17a, it can be confirmed that the compound P-PEG8-Doxo in the form of an albumin-binding prodrug and the compound P-PEG8-Doxo-HSA in the form of an albumin-drug conjugate exhibit excellent anticancer activity against the MDA MB 231 cell line.

[0230] Referring to Figure 17b, it can be confirmed that the compound P-PEG8-MMAE in the form of an albumin-binding prodrug and the compound P-PEG8-MMAE-HSA in the form of an albumin-drug conjugate exhibit excellent anticancer activity against the MDA MB 231 cell line.

[0231] Referring to Figure 17c, it can be confirmed that the compound P-PEG8-Pac in the form of an albumin-binding prodrug and the compound P-PEG8-Pac-HSA in the form of an albumin-drug conjugate exhibit excellent anticancer activity against the MDA MB 231 cell line.

[0232] Referring to FIG. 17d, it can be confirmed that the compound P-PEG8-SN38 in the form of an albumin-binding prodrug and the compound P-PEG8-SN38-HSA in the form of an albumin-drug conjugate exhibit excellent anticancer activity against the MDA MB 231 cell line.

[0233] Evaluation Example 4

[0234] Figures 18a to 18d are drawings showing the results of repeating the evaluation example 2 above, except that the evaluation compounds were changed to compound 31A (P-3C-SS-SN38), compound 31B (P-3C-SS-Pac), compound 31C (P-3C-SS-Doxo), and compound 31D (P-3C-SS-MMAE), which were synthesized using a click reaction.

[0235] Referring to Figure 18a, it can be confirmed that the compound P-3C-SS-Doxo in the form of an albumin-binding prodrug and the compound P-3C-SS-Doxo-HSA in the form of an albumin-drug conjugate exhibit excellent anticancer activity against the HeLA cell line.

[0236] Referring to Figure 18b, it can be confirmed that the compound P-3C-SS-MMAE in the form of an albumin-binding prodrug and the compound P-3C-SS-MMAE-HSA in the form of an albumin-drug conjugate exhibit excellent anticancer activity against the HeLA cell line.

[0237] Referring to FIG. 18c, it can be confirmed that the compound P-3C-SS-Pac in the form of an albumin-binding prodrug and the compound P-3C-SS-Pac-HSA in the form of an albumin-drug conjugate exhibit excellent anticancer activity against the HeLa cell line.

[0238] Referring to FIG. 18d, it can be confirmed that compound P-3C-SS-SN38 in the form of an albumin-binding prodrug and compound P-3C-SS-SN38-HSA in the form of an albumin-drug conjugate exhibit excellent anticancer activity against the HeLA cell line.

[0239] Evaluation Example 5

[0240] Figures 19a to 19d are drawings showing the results of repeating the evaluation example 4 above by changing the HeLA cell line to the MDA MB 231 cell line.

[0241] Referring to FIG. 19a, it can be confirmed that the compound P-3C-SS-Doxo in the form of an albumin-binding prodrug and the compound P-3C-SS-Doxo-HSA in the form of an albumin-drug conjugate exhibit excellent anticancer activity against the MDA MB 231 cell line.

[0242] Referring to Figure 19b, it can be confirmed that the compound P-3C-SS-MMAE in the form of an albumin-binding prodrug and the compound P-3C-SS-MMAE-HSA in the form of an albumin-drug conjugate exhibit excellent anticancer activity against the MDA MB 231 cell line.

[0243] Referring to Figure 19c, it can be confirmed that the compound P-3C-SS-Pac in the form of an albumin-binding prodrug and the compound P-3C-SS-Pac-HSA in the form of an albumin-drug conjugate exhibit excellent anticancer activity against the MDA MB 231 cell line.

[0244] Referring to FIG. 19d, it can be confirmed that compound P-3C-SS-SN38 in the form of an albumin-binding prodrug and compound P-3C-SS-SN38-HSA in the form of an albumin-drug conjugate exhibit excellent anticancer activity against the MDA MB 231 cell line.

[0245] Referring to FIGS. 15a, 15b and 16a to 16d and 18a to 18d together, it can be seen that P-SS-SN38, P-SS-Pac, P-PEG8-MMAE, P-PEG8-Pac, P-PEG8-SN38, P-3C-SS-MMAE, P-3C-SS-Pac, P-3C-SS-SN38 and albumin conjugates thereof effectively exhibit excellent anticancer activity against the HeLa cell line.

[0246] Referring to FIGS. 15c, 15d and 17a to 17d and 19a to 19d together, it can be seen that P-PEG8-MMAE, P-PEG8-SN38, P-3C-SS-MMAE, P-3C-SS-SN38 and their albumin conjugates effectively exhibit excellent anticancer activity against the MDA MB 231 cell line.

[0247] Evaluation Example 6

[0248] Figure 20 is a drawing showing the results of an in vitro anticancer activity evaluation using a FLIC-drug conjugate in which each anticancer drug (doxorubicin, MMAE, paclitaxel, SN38) is bound to FLIC for the HCT116 cell line.

[0249] Referring to Figure 20, it can be confirmed that the albumin-drug conjugate using FLIC exhibits anticancer activity against the HCT116 cell line, and in particular, it can be seen that P-PEG8-SS-SN38-HSA effectively exhibits excellent anticancer activity.

[0250] Referring to Evaluation Examples 1 to 6, since the anticancer effect may differ depending on the type of FLIC conjugate for each anticancer drug, it is necessary to select an appropriate type of FLIC-drug conjugate depending on the type of anticancer drug.

[0251] MMAE anticancer agent is a peptide compound that is rapidly excreted in the body and is highly likely to be hydrolyzed, which may reduce its anticancer effect when applied to actual treatment. However, if it binds to albumin through FLIC, its stability in the blood is improved and the possibility of accumulating in cancer cells increases, so it can be applied to the treatment of various cancers.

[0252] [In vivo anticancer activity evaluation]

[0253] Figure 21 is a schematic diagram of an in vivo anticancer activity evaluation in mice transplanted with the MDA MB 231 cell line. P-PEG8-Doxo and P-PEG8-SN38 were used as FLIC-drug conjugates. As described above, P-PEG8-SN38 exhibited superior anticancer activity than P-PEG8-Doxo in the in vitro anticancer activity evaluation.

[0254] In the in vivo anticancer evaluation, 35 mice were selected from 7 groups, 5 mice each, and the MDA MB 231 cell line was transplanted and grown. Each group was intravenously administered 3 times at 1-week intervals with each of Vehicle, Drug 1 (doxorubicin), Drug 1-1 (P-PEG8-Doxo), Drug 1-2 (HSA-P-PEG8-Doxo), Drug 2 (SN38), Drug 2-1 (P-PEG8-SN38), and Drug 2-2 (HSA-P-PEG8-SN38), and the anticancer effect was confirmed by measuring the weight and volume of the tumor tissues after resecting them.

[0255] Figures 22A to 22D are diagrams showing the results of in vivo anticancer activity evaluation. Specifically, Figure 22A shows the relative change in the average body weight of mice belonging to each group, Figure 22B shows the relative tumor volume of mice belonging to each group, Figure 22C shows the external appearance of mice belonging to each group and photographs of cancer tumors cut from each mouse belonging to each group, and Figure 22D shows the average cancer tumor weight relative to the average body weight of mice belonging to each group.

[0256] Referring to Figures 22a to 22d, it can be confirmed that when Drug 2-2 (HSA-P-PEG8-SN38) was applied, the volume and weight of the cancer tumor were effectively reduced. From this, it can be seen that the FLIC-drug conjugate form, such as Drug 2-2 (HSA-P-PEG8-SN38), is more useful than the anticancer drug itself, such as Drug 2 (SN38).

[0257] [synthesis]

[0258] Reaction Scheme 1 (Compound 4)

[0259]

[0260] Compound 4 is prepared according to the above reaction scheme 1 (KOAc: potassium acetate, PdCl2(dppf): bis(diphenylphosphino)ferrocene)palladium(II) dichloride, DMSO: dimethyl sulfoxide, NBS: N-bromosuccinimide, AIBN: azobisisobutyronitrile).

[0261] Reaction Scheme 2 (Compound 6)

[0262]

[0263] 2-Aminobenzaldehyde (compound 5, 1.00 g, 8.83 mmol) is dissolved in anhydrous acetonitrile (CH3CN, 20 mL) under a nitrogen stream. Potassium carbonate (K2CO3, 2.44 g, 17.66 mmol, 2.0 equiv) is added, and the mixture is stirred at room temperature (rt) for 15 minutes. Then, compound 4 (2.10 g, 8.83 mmol, 1.0 equiv) is added, and the reaction mixture is stirred at room temperature until the reaction is completed, as monitored by thin layer chromatography (TLC). The reaction mixture is filtered to remove inorganic salts, and the filtrate is concentrated under reduced pressure. The product is dried in vacuo to obtain compound 6 (P-NO2) as a solid (2.12 g, 67% yield). The results of NMR analysis for compound 6 are as follows.

[0264] 1 H-NMR (300 MHz, CDCl3): 9.89 (s, 1H), 8.89 (t,J= 9 Hz 1H), 8.87 (s, 1H), 8.71 (d,J= 15 Hz 1H), 8.19 (d,J= 15 Hz 1H), 7.48 (t,J= 3 Hz) 1H), 6.74 (t,J= 6 Hz 1H), 6.60 (d,J= 9 Hz 1H);

[0265] 13C-NMR (100 MHz, CDCl3): 195.08, 149.77, 148.38, 145.54, 137.36, 136.47, 135.33, 121.32, 118.86, 116.06, 111.56, 48.92, 25.32.

[0266] Reaction Scheme 3 (Compound 7)

[0267]

[0268] Compound 6 (P-NO2, 0.500 g, 1.30 mmol) is dissolved in ethanol:water (volume ratio 5:1, 50 mL). Iron powder (Fe, 0.289 g, 5.20 mmol, 4 equiv) and ammonium chloride (NH4Cl, 0.139 g, 2.60 mmol, 2 equiv) are added, and the mixture is stirred at room temperature for 6 h. The reaction is monitored by TLC, the mixture is filtered through a pad of Celite to remove the iron catalyst, and the filtrate is concentrated under reduced pressure. The residue is purified by silica gel column chromatography to give compound 7 (P-NH2) as a white solid (0.390 g, 81% yield). The results of NMR analysis for compound 7 are as follows.

[0269] 1 H-NMR (300 MHz, CDCl3): 9.82 (s, 1H), 8.72 (t,J= 3 Hz 1H), 7.46 (d,J= 15 Hz 1H), 7.44 (t,J= 3 Hz 1H), 6.81 (d,J= 9 Hz 1H), 6.74 (d,J= 9 Hz 1H), 6.71 (t,J= 3 Hz 1H);

[0270] 13 C-NMR (100 MHz, CDCl3): 194.89, 162.32, 150.08, 139.90, 137.34, 136.41, 126.63, 125.23, 124.44, 118.90, 115.75, 111.47, 74.07, 25.32.

[0271] Reaction Scheme 4 (Compound 9)

[0272]

[0273] The above compound 8 (0.470 g, 1.70 mmol), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI in the form of EDC·HCl, 0.70 g, 3.00 mmol), and 4-dimethylaminopyridine (DMAP, 120 mg, 0.70 mmol) are dissolved in anhydrous dichloromethane (DCM, 10 mL) under a nitrogen stream at 0°C. The reaction mixture is stirred for 30 minutes, and then compound 7 (P-NH2, 0.50 g, 1.85 mmol) is added. The mixture is heated to room temperature and stirred vigorously for 12 hours. The reaction progress is monitored by TLC (silica gel, monitored by a UV lamp). After completion of the reaction, the solvent was removed under reduced pressure, and the product was purified by silica gel column chromatography (methylene chloride / methanol, 95:5 v / v) to obtain compound 9 (P-SS-COOH) as a pale yellow solid (0.58 g, 61% yield). The NMR analysis results for compound 9 are as follows.

[0274] 1 H-NMR (300 MHz, CDCl3): 9.77 (s, 1H), 8.01 (s, 1H), 7.82 (d, 3Hz, 1H), 7.75 (d, 3Hz, 1H), 7.45 (d, 3Hz, 1H), 7.35 f 7.82 (m, 2H), 6.75 (d, 9Hz, 1H), 6.67 (t, 9Hz, 1H)

[0275] 13 C-NMR (100 MHz, CDCl3): 194.68, 171.05, 150.16, 137.53, 136.30, 135.11, 129.78, 128.39, 124.55, 118.58, 115.32, 111.70, 45.86, 24.95

[0276] [General Synthesis Example A (Synthesis of Ac-CL-D)]

[0277] Compound 9 (P-SS-COOH), EDCI, and DMAP were dissolved in anhydrous DCM (5 mL) at room temperature under a nitrogen atmosphere. The reaction mixture was stirred for 30 minutes, after which each anticancer agent was added. The mixture was stirred vigorously at room temperature for 12 hours, and the reaction progress was monitored by TLC (silica gel, monitored by a UV lamp). After completion of the above process, the solvent was removed under reduced pressure, and the product was purified by silica gel column chromatography (methylene chloride / methanol, 95:5 v / v) to obtain the desired complex.

[0278] Synthetic Example A1 (Synthesis of P-SS-SN38): Compounds 11 and 12

[0279]

[0280] Compound 9 (P-SS-COOH, 0.10 g, 0.153 mmol), EDCI (0.40 g, 1.33 mmol), and DMAP (0.20 mL, 0.14 mmol) are dissolved in DCM (5 mL). SN38 (0.150 g, 4.20 mmol) is added and the general synthesis method A is followed. Compound 11 (albumin-binding prodrug, P-SS-SN38) is obtained as a pale yellow solid (0.09 g, 43% yield) after purification. Since some of the albumin-binding moieties of compound 11 are protected with pinacol to form pinacolborane, it can be referred to as P-SS-SN38_protected. The results of NMR analysis for compound 11 are as follows.

[0281] 1H-NMR (300 MHz, CDCl3): 9.83 (s, 1H), 8.77 (m, 1H), 8.25 (s, 1H), 8.22 (s, 1H), 7.11 (s, 1H), 7.71 (s, 1H) ,7.66 (s, 1H), 7.59 (d, 9Hz, 1H), 7.48 (d, 9Hz, 1H), 7.40-7.25 (m, 4H), 6.73-6.65 (m, 2H), 5.80 (d, 18Hz, 1H), 5.35 (d, 18Hz, 1H), 5.26 (s, 2H), 4.72-4.64 (m, 2H), 3.83 (s, 1H), 3.16-3.11(m, 2H), 2.83-2.80 (m, 6H), 2.25-2.48 (m, 2H), 2.27-2.13 (m, 4H), 1.93-1.89 (m, 2H), 1.64 (s, 3H), 1.40(t, 9Hz, 3H), 1.34 (m, 12H), 1.06 (t, 9Hz, 3H).

[0282] 13 C-NMR (100 MHz, CDCl3): 193.90, 173.94, 171.64, 170.51, 157.68, 150.85, 150.31, 145.39, 138.04, 136.66, 135.86, 127.36, 125.40, 118.65, 114.99, 114.66, 111.59, 98.13, 83.83, 66.38, 49.472, 46.25, 37.608, 32.70, 31.68, 24.916, 24.449,14.06, 7.09

[0283] Compound 12, which is formed by removing pinacol from the above compound 11 in the presence of albumin in an aqueous solution, can be referred to as P-SS-SN38_deprotected, as it is not protected by pinacol. The results of NMR analysis for compound 12 are as follows.

[0284] 1H-NMR (300 MHz, CDCl3): 9.70 (s, 1H), 8.08 (d, 9Hz, 1H), 7.82 (s, 1H), 7.53-7.51 (m, 2H), 7.34-7.25 (m, 3H), 6.73-6.60 (m, 3H), 5.36 (s, 2H), 5.12 (s, 2H), 4.24 (s, 2H), 3.18-3.10 (m, 2H), 2.81-2.68 (m, 4H), 2.34-2.25 (m, 2H), 2.08-2.98 (m, 2H), 1.89-1.87 (m, 2H), 1.78-1.68 (m, 4H), 1.23 (t, 3H), 0.82 (t,9Hz, 3H)

[0285] 13 C-NMR (100 MHz, CDCl3): 194.18, 173.84, 117.17 170.43, 160.93, 157.65, 158.15, 151.47, 149.80, 147.01, 145.62, 136.71, 131.98, 130.86, 127.80, 125.29, 120.84, 118.33, 116.38, 114.44, 111.93, 98.29, 72.83, 65.71, 62.93, 49.84, 42.88, 38.15, 35.36, 32.88, 31.74, 29.71, 25.06, 23.65, 13.97, 7.83

[0286] Synthesis Example A2 (Synthesis of P-SS-Pac): Compounds 14 and 15

[0287]

[0288] Compound 9 (P-SS-COOH, 0.10 g, 0.153 mmol), EDCI (0.20 g, 1.33 mmol), and DMAP (0.30 mL, 0.14 mmol) are dissolved in DCM (5 mL). Paclitaxel (Pac, 0.250 g, 4.20 mmol) is added and General Synthesis Example A is followed. Purification yields compound 14 (albumin-binding prodrug, P-SS-Pac) as a white solid (0.11 g, 47% yield). Since some of the albumin-binding moieties of compound 14 are protected with pinacol to form pinacolborane, it can be referred to as P-SS-Pac_protected. The results of NMR analysis for compound 14 are as follows.

[0289] 1 H-NMR (300 MHz, CDCl3): 9.78 (s, 1H), 8.75 (t, 6Hz, 1H), 8.17 (d, 6Hz, 1H), 7.75-7.60 (m, 4H), 7.56-7.48 (m, 4H), 7.46-7.29 (m, 15H) 7.11 (d, 9Hz, 1H), 6.68-6.63 (m, 2H), 6.32 (s, 1H), 6.25 (t, 9Hz, 1H), 6.00 (d, 9Hz, 1H), 5.71 (d, 12Hz, 1H), 5.54 (t, 3Hz, 1H), 5.00 (d, 9Hz, 1H), 4.75 (bs, 2H), 4.69 (bs, 1H), 4.34 (dd, 18Hz,2Hz 2H), 3.84 (d, 9Hz, 1H), 2.68-2.48 (m, 12H), 2.27 (s, 3H), 1.95 (s, 3H), 1.70 (s, 3H), 1.32 (s, 12H), 1.22 (s, 3H), 1.28 (s, 3H), 1.14 (s, 3H)

[0290] 13C-NMR (100 MHz, CDCl3): 203.87, 193.93, 172.14, 171.32, 170.59, 169.87, 168.22, 167.35, 167.29, 167.04, 167.70, 150.83, 146.40, 140.78, 135.89, 132.09, 129.29, 129.29, 129.19, 128.65, 126.79, 118.49, 117.71, 117.34, 115.00, 111.63, 83.84, 81.10, 79.17, 79.15, 77.43, 75.14, 74.16, 72.22, 71.92, 58.57, 53.05, 46.24, 45.69, 43.26, 37.84, 35.65, 29.81, 24.91, 24.11, 22.81, 20.96, 14.68, 9.74

[0291] Compound 15, which is formed by removing pinacol in the presence of albumin in an aqueous solution of the above compound 14, can be referred to as P-SS-Pac_deprotected, as it is not protected by pinacol. The results of NMR analysis for compound 15 are as follows.

[0292] 1H-NMR (300 MHz, CDCl3): 9.79 (s, 1H), 8.58 (t, 6Hz, 1H), 8.14 (d, 6Hz, 1H), 7.76 (m, 4H), 7.81-7.61 (m, 4H), 7.59-7.29 (m, 15H), 7.13 (d, 9Hz, 1H), 6.68-6.63 (m, 2H), 6.32 (s, 1H), 6.21 (t, 9Hz, 1H), 6.08 (d, 9Hz, 1H), 5.73 (d, 12Hz, 1H), 5.56 (t, 3Hz, 1H), 5.03 (d, 9Hz, 1H), 4.75 (bs, 2H), 4.69 (bs, 1H), 4.34 (dd, 18Hz,2Hz, 2H), 3.84 (d, 9Hz, 1H),2.68-2.48 (m, 12H), 2.27 (s, 3H), 1.93 (s, 3H), 1.71 (s, 3H), 1.22 (s, 3H), 1.28 (s, 3H), 1.14 (s, 3H)

[0293] 13 C-NMR (100 MHz, CDCl3):207.08, 203.80, 193.97, 175.10, 171.28, 169.88, 168.27, 166.96, 150.59, 142.64, 136.91, 133.69, 132.81, 130.21, 129.33, 129.09, 128.74, 127.18, 126.71, 124.59, 99.99, 84.43, 81.05, 79.07, 58.47, 43.16, 30.95, 29.70, 22.90, 20.86, 14.86

[0294] 합성예 A3 (P-SS-Doxo의 합성): 화합물 17 및 18

[0295]

[0296] Compound 9 (P-SS-COOH, 0.10 g, 0.153 mmol), EDCI (0.18 g, 0.17 mmol), and DMAP (0.20 mL, 0.14 mmol) are dissolved in DCM (5 mL). Doxorubicin·HCl (0.150 g, 0.17 mmol) is added and the general synthesis method A is followed. Purification yields compound 17 (albumin-binding prodrug, P-SS-Doxo) as a red solid (0.08 g, 44% yield). Since some of the albumin-binding moieties of compound 17 are protected with pinacol to form pinacolborane, it can be referred to as P-SS-Doxo_protected. The NMR analysis results for compound 17 are as follows.

[0297] 1 H-NMR (300 MHz, CDCl3): 13.95 (s, 1H), 13.21 (s, 1H), 9.80 (s, 1H), 8.27 (s, 1H), 8.03 (d, 9Hz, 1H), 7.75-7.60 (m, 3H), 7.43-7.28 (m, 2H), 7.21 (d, 9Hz, 1H), 6.73 (d, 9Hz, 1H), 6.68 (t, 6Hz, 1H), 6.22 (bs, 1H), 5.47 (s, 1H), 5.23 (s, 1H), 4.76 (s, 1H), 4.57 (s, 1H), 4.17-4.05 (m, 4H), 3.64 (s, 1H), 3.26-2.69 (m, 7H), 2.48-1.87 (m, 16H), 1.33 (s, 12H), 1.27-1.25 (m, 6H)

[0298] 13C-NMR (100 MHz, CDCl3): 214.23, 194.28, 187.37, 172.10, 171.00, 161.30, 156.37, 151.04, 141.62, 136.91, 134.69, 130.81, 128.87, 127.48, 123.05, 121.39, 119.72. 118.34, 114.74, 111.16, 83.70, 69.59, 67.64, 65.42, 56.54, 46.28, 45.17, 38.52, 35.75, 34.64, 32.15, 30.21, 24.94, 22.73, 21.62, 17.46, 14.41

[0299] Compound 18, which is formed by removing pinacol in the presence of albumin in aqueous solution from the above compound 17, can be referred to as P-SS-Doxo_deprotected, as it is not protected by pinacol. The results of NMR analysis for compound 18 are as follows.

[0300] 1 H-NMR (300 MHz, CDCl3): 13.95 (s, 1H), 13.21 (s, 1H), 9.74 (s, 1H), 7.81-7.68 (m, 4H), 7.60-7.34 (m, 7H), 7.21 (d, 9Hz, 1H), 7.01 (d, 9Hz, 1H), 6.70 (d, 9Hz, 1H), 6.64 (t, 6Hz, 1H), 5.18 (s, 2H), 4.81 (s, 2H), 4.57 (s, 3H), 4.47 (s, 1H), 4.15-4.13 (m, 2H), 3.91-3.81 (m, 9H), 3.39 (m, 3H), 2.81-2.57 (m, 14H), 2.53-1.74 (m, 20H), 1.43-1.40 (m, 2H), 1.61-1.06 (m, 12H)

[0301] 13C-NMR (100 MHz, CDCl3): 214.51, 194.83, 186.52, 171.83, 170.72, 161.02, 156.31, 154.64, 150.78, 138.02, 136.35, 133.86, 129.98, 120.56, 116.62. 111.41, 101.16, 75.66, 70.12, 68.45, 67.07, 65.30, 57.37, 49.05, 35.20, 25.22, 21.62, 17.18.

[0302] Reaction Scheme 5 (Compound 19)

[0303]

[0304] Bis-PEG8 (1.70 g, 3.60 mmol), EDC·HCl (0.70 g, 5.20 mmol), and DMAP (110 mg, 0.90 mmol) are dissolved in anhydrous dichloromethane (DCM, 10 mL) under a nitrogen stream at 0°C. The reaction mixture is stirred for 30 min, and then compound 7 (P-NH2, 0.50 g, 1.85 mmol) is added. The mixture is heated to room temperature and stirred vigorously for 12 h. The reaction progress is monitored by TLC (silica gel, monitored by a UV lamp). After completion of the above process, the solvent is removed under reduced pressure, and the residue is purified by silica gel column chromatography (methylene chloride / methanol, 95:5 v / v) to obtain compound 19 (P-PEG8) as a colorless solid (1.15 g, 57% yield). The NMR analysis results for compound 19 are as follows.

[0305] 1H-NMR (300 MHz, CDCl3): 9.80 (s, 1H), 8.75 (t, 3Hz, 1H), 8.65 (s, 1H), 7.81-7.35 (m, 2H), 7.35 (d, 9Hz, 1H), 7.30-7.28 (m, 2H), 6.74 (d, 9Hz, 1H), 6.64 (t, 9Hz, 1H), 4.66 (s, 2H), 3.67-3.61 (m, 4H), 3.60-3.55(m, 28H), 2.85-2.58 (m, 4H), 1.1314 (s, 12H).

[0306] 13 C-NMR (100 MHz, CDCl3):193.74, 174.40, 170.26, 150.73, 140.41, 136.65, 135.63, 128.43, 128.01, 122.94, 118.36, 114.74, 111.47, 83.91, 70.45, 70.25, 67.10, 66.55, 45.96, 37.77, 34.90, 29.67, 24.85.

[0307] 반응식 6 (화합물 20)

[0308]

[0309] Compound 19 (P-PEG8, 0.59 g, 1.33 mmol), EDC·HCl (0.308 g, 2.95 mmol), and DMAP (80 mg, 0.14 mmol) are dissolved in anhydrous dichloromethane (DCM, 10 mL) at 0 °C under a nitrogen stream. The reaction mixture is stirred for 30 min, and then HO-CH2CH2-SS-CH2CH2-OH (0.824 g, 4.20 mmol) is added. The mixture is heated to room temperature and stirred vigorously for 12 h. The reaction progress is monitored by TLC (silica gel, monitored by a UV lamp). After completing the above process, the solvent is removed under reduced pressure, and the residue is purified by silica gel column chromatography (methylene chloride / methanol, 95:5 v / v) to obtain compound 20 (P-PEG8-SS-OH) as a colorless solid (0.36 g, 47% yield). The NMR analysis results for compound 20 are as follows.

[0310] 1 H-NMR (300 MHz, CDCl3): 9.81 (s, 1H), 8.76 (t, 9Hz, 1H), 8.59 (s, 1H), 7.81-7.35 (m, 2H), 7.34 (d, 9Hz, 1H), 7.31-7.29 (m, 2H), 6.74 (d, 9Hz, 1H), 6.65 (t, 9Hz, 1H), 4.66 (d, 9Hz, 2H), 4.39 (t, 8Hz, 2H), 3.68-3.63 (m, 4H), 3.60-3.57 (m, 28H), 2.90-2.86 (m, 5H), 2.64-2.60 (m, 6H), 2.01-2.07 (m, 2H), 1.32 (s, 12H).

[0311] 13C-NMR (100 MHz, CDCl3): 193.17, 171.48, 170.07, 150.73, 140.40, 136.73, 136.63, 135.65, 128.42, 127.96, 122.91, 118.40, 114.73, 111.48, 83.92, 70.49, 70.37, 66.50, 62.42, 60.27, 45.97, 41.52, 37.90, 37.04, 35.00, 29.70, 24.86.

[0312] Reaction Scheme 7 (Compound 22)

[0313]

[0314] Compound 20 (P-PEG8-SS-OH, 0.470 g, 1.70 mmol), EDC·HCl (0.50 g, 3.00 mmol), and DMAP (0.12 g, 0.70 mmol) are dissolved in anhydrous dichloromethane (DCM, 10 mL) at 0 °C under a nitrogen stream. The reaction mixture is stirred for 30 min, and then compound 21 (p-nitrophenyl chloroformate, 0.300 g, 1.85 mmol) is added. The mixture is heated to room temperature and stirred vigorously for 12 h. The reaction progress is monitored by TLC (silica gel, monitored by a UV lamp). After completing the above process, the solvent is removed under reduced pressure, and the product is purified by silica gel column chromatography (methylene chloride / methanol, 95:5 v / v) to obtain compound 22 (P-PEG8-SS-NO2) as a pale yellow solid (0.34 g, yield 53%). The NMR analysis results for compound 22 are as follows.

[0315] 1H-NMR (300 MHz, CDCl3): 10.06 (s, 1H), 8.65 (s, 1H), 8.27 (d, 9Hz, 2H), 8.20 (d, 9Hz, 1H), 7.86 (d, 9Hz, 1H), 7.71 (s, 1H), 7.51-7.38 (m, 5H), 7.21 (d, 9Hz, 1H), 7.00 (d, 9Hz, 1H), 5.46 (d, 15Hz, 1H), 5.20 (d, 15Hz, 1H), 4.45 (t, 6Hz, 2H), 4.37 (t, 6Hz, 2H), 3.83 (t, 6Hz, 2H), 3.74 (t, 6Hz, 2H), 3.69-3.60 (m, 26H), 3.03 (t, 9Hz, 2H), 2.96 (t, 6Hz, 2H), 2.66-2.59 (m, 4H), 1.25 (s, 4H), 1.15-1.12 (m, 12H).

[0316] 13 C-NMR (100 MHz, CDCl3): 190.02, 171.42, 170.20, 155.47, 153.14, 152.38, 152.37, 145.54, 144.98, 141.55, 134.64, 126.13, 125.43, 125.41, 125.40, 125.39, 125.06, 123.33, 122.30, 121.89, 83.81, 70.51, 66.52, 62.32, 37.99, 37.18, 36.76, 35.00, 29.76, 24.88, 24.68.

[0317] [일반 합성예 B (Ac-SL-CL-D의 합성)]

[0318] Compound 22 (P-PEG8-SS-NO2) and triethylamine (TEA) were dissolved in anhydrous DMF (5 mL) under a nitrogen atmosphere at room temperature. The reaction mixture was stirred for 30 minutes, and then each anticancer agent was added. The mixture was stirred vigorously at room temperature for 12 hours, and the reaction progress was monitored by TLC (silica gel, monitored by a UV lamp). After the above process was completed, the solvent was removed under reduced pressure, and the crude product was purified by silica gel column chromatography (methylene chloride / methanol, 95:5 v / v) to obtain the desired complex.

[0319] Synthesis Example B1 (Synthesis of P-PEG8-SS-SN38): Compound 23

[0320]

[0321] Compound 22 (P-PEG8-SS-NO2, 0.40 g, 1.33 mmol) and TEA (0.20 mL, 0.14 mmol) are dissolved in DMF (5 mL). SN38 (0.150 g, 4.20 mmol) is added and the general synthesis example B is followed. The result of purification is compound 23 (P-PEG8-SS-SN38, 0.21 g, 49% yield) as a pale yellow solid. The results of NMR analysis for compound 23 are as follows.

[0322] 1H-NMR (300 MHz, CDCl3): 10.09 (s, 1H), 8.65 (s, 1H), 8.28 (d, 9Hz, 2H), 8.21 (d, 9Hz, 1H) 7.94 (s, 1H), 7.89 (d, 6Hz, 2H), 7.71-7.65 (m, 3H), 7.51-7.42 (m, 3H), 7.22 (d, 9Hz, 2H), 7.00 (d, 6Hz, 1H), 5.78 (d, 15Hz, 1H), 5.47 (d, 15Hz, 1H), 5.34-5.27 (m, 4H), 5.21 (d, 15Hz, 1H), 4.59 (t, 6Hz, 2H), 4.40 (t, 6Hz, 2H), 4.02-4.00 (m, 1H), 3.83 (t, 6Hz, 2H), 3.76 (t, 6Hz, 2H), 3.72-3.52 (m, 32H), 3.20-3.16 (m, 2H), 3.09 (t, 6Hz, 2H), 2.99 (t, 6Hz, 2H), 2.89 (s, 1H), 2.67-2.61 (m, 4H), 2.18-2.06 (m, 2H), 1.96-1.86 (m, 2H), 1.41 (t, 6Hz, 3H), 1.26-1.25 (m, 3H), 1.16-1.13(m, 12H), 1.04 (t, 6Hz, 3H).

[0323] 13 C-NMR (100 MHz, CDCl3): 190.00, 173.90, 171.43, 170.166, 162.631, 157.68, 153.27, 150.28, 149.80, 145.50, 141.57, 137.65, 132.36, 127.49,125.06, 124.71, 122.28, 118.77, 114.21, 98.19, 83.81, 70.58, 70.46, 66.52, 62.34, 52.42, 49.46, 37.19, 36.90, 35.01, 31.68, 29.75, 24.88, 24.68, 23.26, 14.07, 7.91.

[0324] 합성예 B2 (P-PEG8-SS-Pac의 합성): 화합물 24

[0325]

[0326] Compound 22 (P-PEG8-SS-NO2, 0.20 g, 1.33 mmol) and TEA (0.30 mL, 0.14 mmol) are dissolved in DMF (5 mL). Paclitaxel (Pac, 0.250 g, 4.20 mmol) is added and general synthesis example B is followed. Purification results in the form of a white solid, compound 24 (P-PEG8-SS-Pac, 0.18 g, yield 41%). The NMR analysis results for compound 24 are as follows.

[0327] 1 H-NMR (300 MHz, CDCl3): 10.08 (s, 1H), 8.63 (s, 1H), 8.20-8.13 (m, 4H), 7.87 (d, 9Hz, 2H), 7.77 (d, 9Hz, 2H), 7.70 (s, 1H), 7.61-7.35 (m, 15H), 7.21-7.13 (m, 3H), 7.00 (d, 9Hz, 1H), 6.30-6.25 (m, 2H), 6.02 (d, 6Hz, 1H), 5.70 (d, 6Hz, 1H), 5.47-5.40 (m, 2H), 5.20 (d, 15Hz, 1H), 4.99 (d, 15Hz, 1H), 4.40 (t, 6Hz, 2H), 4.30 (t, 6Hz, 2H), 4.22 (d, 9Hz, 1H), 3.81 (t, 6Hz, 2H), 3.74-3.59 (m, 34H), 2.94-2.86 (m, 4H), 2.64-2.56 (m, 6H), 2.49-2.38 (m, 5H), 2.39-2.23 (m, 6H), 2.20-1.80 (m, 5H), 1.69 (s, 3H), 1.26-1.24 (m, 6H), 1.15-1.125 (m, 16H).

[0328] 13C-NMR (100 MHz, CDCl3): 203.87, 190.03, 171.48, 171.28, 169.96, 167.93, 167.33, 167.02, 156.07, 154.03, 153.14, 144.97, 142.61, 141.55, 136.75, 132.07, 129.30, 129.19, 128.81, 128.79, 128.62, 127.30, 126.75, 125.07, 122.29, 84.50, 83.87, 81.21, 78.79, 76.11, 75.11, 72.13, 70.58, 66.36, 62.36, 58.56, 52.84, 52.42, 45.67, 43.28, 35.66, 34.99, 26.88, 24.89, 24.69, 22.80, 20.91, 14.88, 14.20, 9.69.

[0329] Synthesis Example B3 (Synthesis of P-PEG8-SS-Doxo): Compound 25

[0330] Compound 22 (P-PEG8-SS-NO2, 0.18 g, 0.17 mmol) and TEA (0.20 mL, 0.14 mmol) are dissolved in DMF (5 mL). Doxorubicin·HCl (0.150 g, 0.17 mmol) is added and general synthesis example B is followed. As a result of purification, compound 25 (P-PEG8-SS-Doxo, 0.11 g, yield 43%) is obtained as a red solid.

[0331] [General Synthesis Example C (Synthesis of Ac-SL-D): SL = PEG8]

[0332] The reactions mentioned in General Synthesis Example C are carried out under a nitrogen atmosphere at room temperature (rt) unless otherwise specified. The progress of the reaction was monitored by chromatography (TLC) on silica gel plates (confirmed by ultraviolet light or appropriate staining), and column chromatography was performed on silica gel (230-400 mesh) using methylene chloride / methanol (MC / MeOH) as the eluent.

[0333] DMAP and EDCI are added to anhydrous DCM (5 mL) in the amounts indicated below to a solution of compound 19 (P-PEG8). The mixture is stirred at room temperature for 30 minutes under nitrogen. Each anticancer drug (SN38, paclitaxel, doxorubicin, monomethyl auristatin E) is then added, and the reaction mixture is stirred vigorously at room temperature for 12 hours. After completion of the above process, the mixture is monitored by TLC, the solvent is evaporated under reduced pressure, and the product is purified by silica gel column chromatography (MC / MeOH, 95:5 v / v) to obtain the desired complex.

[0334] Synthesis Example C1 (Synthesis of P-PEG8-SN38): Compound 26A

[0335]

[0336] Compound 19 (P-PEG8, 0.16 g, 0.20 mmol), DMAP (0.05 g, 0.20 mmol), and EDC·HCl (0.077 g, 0.40 mmol) are dissolved in DCM (5 mL). After stirring for 30 min, SN38 (0.078 g, 0.20 mmol) is added, and the reaction mixture is stirred at room temperature for 12 h. As a result of purification, compound 26A (P-PEG8-SN38, 0.12 g, 51% yield) is obtained as a pale yellow solid. The NMR analysis results for compound 26A are as follows.

[0337] 1H-NMR (300 MHz, CDCl3): 9.76 (s, 1H), 8.70 (s, 2H), 8.21 (d, 9Hz, 2H), 7.98 (s, 2H), 7.79-7.71 (m, 3H), 7.53-7.38 (m, 2H), 7.53-7.38 (m, 2H), 6.70 (d, 6Hz, 1H), 6.60 (t, 6Hz, 1H), 5.71 (d, 15Hz, 1H), 5.27-5.19 (m, 3H), 4.62 (d, 6Hz, 2H), 3.89 (t, 6Hz, 6Hz, 3.6Hz), t 2H), 3.67-3.54 (m, 39H), 3.25-3.22 (m, 2H), 3.11-3.08 (m, 2H), 2.66 (t, 6Hz, 3H), 2.25-2.11 (m, 3Hz), 6Hz, 3H), 1.32-1.20 (m, 13H), 0.96 (t, 6Hz, 3H).

[0338] 13 C-NMR (100 MHz, CDCl3): 194.72, 173.83, 170.16, 162.64, 157.66, 150.76, 150.35, 149.64, 140.40, 136.66, 13.7, 2.7, 2.7. 127.99, 118.45, 114.78, 111.52, 98.19, 83.97, 73.11, 67.18, 66.42, 66.30, 49.44, 49.32, 46.02, 37.96, 3.35, 3.8, 3.6 29.73, 24.91, 17.92, 14.18, 14.04, 7.92.

[0339] 합성예 C2 (P-PEG8-Pac의 합성): 화합물 26B

[0340]

[0341] Compound 19 (P-PEG8, 0.16 g, 0.133 mmol), DMAP (0.10 g, 0.10 mmol), and EDC·HCl (0.90 g, 2.14 mmol) are dissolved in DCM (5 mL). After stirring for 30 min, paclitaxel (Pac, 0.170 g, 0.20 mmol) is added, and the reaction mixture is stirred at room temperature for 12 h. Through the purification process, compound 26B (P-PEG8-Pac, 0.18 g, 63% yield) is obtained as a white solid. The NMR analysis results for compound 26B are as follows.

[0342] 1 H-NMR (300 MHz, CDCl3): 9.82 (s, 1H), 9.00 (s, 1H), 8.68 (s, 1H), 8.13 (d, 9Hz, 1H), 7.79 (d, 9Hz, 1H), 7.59-7.23 (m, 16H), 6.68 (t, 6Hz, 1H), 6.63 (d, 9Hz, 1H), 6.30 (s, 1H), 6.20 (t, 9Hz, 1H), 5.95-5.91 (m, 1H), 5.69 (d, 6Hz, 1H), 5.51 (d, 6Hz, 1H), 4.97 (d, 9Hz, 1H), 4.46-4.40 (m, 24H), 4.31 (d, 9Hz, 1H), 4.22 (d, 9Hz, 1H), 3.84 (t, 9Hz, 3H), 3.66-3.51 (m, 27H), 3.59-3.49 (m, 4H), 2.93-2.90 (m, 10H), 2.69-2.59 (m, 4H), 2.41 (s, 3H), 2.40-2.21 (m, 5H), 1.92 (s, 3H), 1.68 (s, 3H), 1.38-1.13 (m, 12H).

[0343] 13C-NMR (100 MHz, CDCl3): 210.88, 203.94, 194.15, 171.10, 170.82, 170.25, 169.91, 168.34, 167.46, 166.84, 150.55, 148.65, 142.55, 130.24, 129.01, 128.60, 127.52, 127.42, 120.34, 118.66, 115.34, 111.45, 84.53, 81.33, 76.97, 70.51, 67.24, 66.25, 58.49, 53.26, 51.72, 46.25, 43.29, 39.25, 34.77, 29.74, 26.82, 22.75, 22.22, 20.91, 19.83, 9.73.

[0344] Synthesis Example C3 (Synthesis of P-PEG8-Doxo): Compound 26C

[0345] Compound 19 (P-PEG8, 0.16 g, 0.133 mmol), DMAP (0.10 g, 0.10 mmol), and EDC·HCl (0.90 g, 2.14 mmol) are dissolved in DMF (5 mL). After stirring for 30 minutes, doxorubicin·HCl (0.115 g, 0.20 mmol) is added, and the reaction mixture is stirred at room temperature for 12 hours. As a result of purification, compound 26C (P-PEG8-Doxo, 0.10 g, 43% yield) is obtained as a red solid. The NMR analysis results for compound 26C are as follows.

[0346] 1H-NMR (300 MHz, CDCl3): 13.95 (s, 1H), 13.23 (s, 1H), 9.81 (s, 1H), 8.77 (s, 1H), 8.73 (s, 1H), 8.03 (d, 9Hz, 1H), 7.82-7.63 (m, 3H), 7.46-7.29 (m, 4H), 6.87 (d, 9Hz, 1H), 6.74 (d, 6Hz, 1H), 6.65 (t, 9Hz, 1H), 5.51 (s, 1H), 5.57 (s, 1H), 4.77-4.65 (m, 4H), 4.41-4.39 (m, 1H), 4.13-4.07 (m, 4H), 3.84-3.59 (m, 42H), 3.28 (m, 1H), 3.01-2.89 (m, 3H), 2.63 (t, 6Hz, 3H), 2.48-2.29 (m, 5H), 2.18-2.17 (m, 1H), 1.91-1.69 (m, 1H), 1.79-1.75 (m, 1H), 1.33-1.26 (m, 14H), 0.89-0.86 (m, 3H).

[0347] 13 C-NMR (100 MHz, CDCl3): 214.148, 193.92, 187.09, 186.68, 171.20, 170.17, 170.00, 161.07, 156.36, 150.79, 140.62, 140.45, 136.68, 135.71, 128.48, 128.01, 119.87, 118.49, 114.81, 111.54, 111.36, 101.05, 83.99, 70.64, 70.55, 70.53, 70.48, 70.37, 56.72, 46.04, 45.35, 37.97, 37.15, 29.76, 29.69, 24.98, 24.92, 17.11, 14.19.

[0348] 합성예 C4 (P-PEG8-MMAE의 합성): 화합물 26D

[0349] Compound 19 (P-PEG8, 0.16 g, 0.133 mmol), DMAP (0.10 g, 0.10 mmol), and EDC·HCl (0.90 g, 2.14 mmol) are dissolved in DMF (5 mL). After stirring for 30 min, monomethyl auristatin E (MMAE, 0.143 g, 0.20 mmol) is added, and the reaction mixture is stirred at room temperature for 12 h. During the purification process, compound 26D (P-PEG8-MMAE, 0.21 g, 67% yield) is obtained as a white solid. The NMR analysis results for compound 26D are as follows.

[0350] 1 H-NMR (300 MHz, CDCl3): 9.77 (s, 1H), 8.76 (t, 6Hz, 1H), 8.59 (s, 1H), 7.81-7.75 (m, 2H), 7.45 (d, 6Hz, 1H), 7.39-7.24 (m, 6H), 6.74 (d, 6Hz, 1H), 6.67-6.55 (m, 2H), 4.68 (d, 9Hz, 2H), 4.62-4.56 (m, 1H), 4.28-4.06 (m, 3H), 3.83-3.76 (m, 6H), 3.67-3.55 (m, 30H), 3.40 (s, 3H), 3.31 (s, 3H), 3.00-2.97 (m, 4H), 2.67-2.61 (m, 4H), 2.03-1.96 (m, 2H), 1.84-1.82 (m, 2H), 1.32 (s, 12H), 1.26 (s, 6H), 1.04-0.81(m, 12H).

[0351] 13C-NMR (100 MHz, CDCl3): 193.74, 174.61, 172.91, 172.17, 170.60, 170.07, 169.80, 169.71, 150.79, 141.36, 140.499, 136.68, 135.69 128.47, 128.02, 122.97, 118.48, 114.79, 111.54, 83.97, 83.52, 82.04, 70.58, 67.18, 60.98, 58.05, 54.03, 51.63, 46.04, 44.98, 37.97, 34.17, 30.74, 29.75, 25.05, 24.98, 24.91, 19.42, 13.95, 11.01.

[0352] Reaction Scheme 8 (Compound 28)

[0353]

[0354] A mixture of compound 27 (11-azido-3,6,9-trioxaundecanoic acid, 0.5 g, 2.14 mmol), DMAP (0.26 g, 0.1 mmol), and EDCI (0.41 g, 2.14 mmol) was dissolved in 5 mL of DMF solution at room temperature under a nitrogen stream, and the mixture was stirred vigorously for 12 h at room temperature. Compound 7 (P-NH2, 0.58 g, 2.2 mmol) was added to the reaction mixture over 30 min, followed by vigorous stirring at room temperature. The reaction progress was monitored using TLC. After completion of the reaction, the reaction solvent was removed, and the product was isolated using a silica gel column (methylene chloride / methanol 5%) with a yield of 49%. The results of NMR analysis for compound 28 are as follows.

[0355] 1H-NMR (300 MHz, CDCl3): 9.82 (s, 1H), 8.81-8.76 (m, 2H), 7.97 (d, 6Hz, 1H), 7.75 (s, 1H), 7.47 (d, 9Hz, 1H), 7.39-7.30 (m, 3H), 6.76 (d, 9Hz, 1H), 6.64 (t, 9Hz, 1H), 4.83 (s, 1H), 4.70 (d, 9Hz, 1H), 4.14-4.09 (s, 3H), 3.76 (s, 6H), 3.72-3.36 (m, 2H), 3.60 (t, 9Hz, 2H), 3.28 (t, 9Hz, 2H), 1.34 (s, 12H).

[0356] 13 C-NMR (100 MHz, CDCl3): 193.73, 169.26, 150.71, 140.99, 136.64, 135.88, 135.65, 128.67, 127.96, 122.88, 118.44, 114.78, 111.46, 84.00, 50.64, 45.97, 24.67.

[0357] Reaction Formula 9 (Click Reaction)

[0358]

[0359] Compound 29 (0.40 g, 1.44 mmol), DMAP (0.017 g, 0.10 mmol), and EDC·HCl (0.52 g, 2.84 mmol) are dissolved in anhydrous DCM (8 mL) at room temperature under a nitrogen stream. The mixture is stirred for 30 min, and then each drug derivative (SN38, Pac, Doxo, MMAE) is added. The reaction mixture is stirred vigorously at room temperature for 12 h. The reaction progress is monitored by TLC (silica gel, monitored by a UV lamp). After completion of the above process, the solvent is removed under reduced pressure, and the crude product is purified by silica gel column chromatography (methylene chloride / methanol, 95:5 v / v) to obtain the desired product.

[0360] Reaction Scheme 9A (Compound 30A)

[0361] Compound 30A was obtained in 89% yield following the general synthesis example D using SN38 (0.56 g, 1.42 mmol) as a drug derivative. The NMR analysis results for compound 30A are as follows.

[0362] 1 H-NMR (300 MHz, CDCl3): 8.26 (d, 9Hz, 1H), 7.91 (s, 1H), 7.81-7.62 (m, 2H), 5.77 (d, 15Hz, 1H), 5.33-5.25 (m, 3H), 4.61 (t, 6Hz, 2H), 4.25 (s, 2H), 4.15 (s, 1H), 3.84 (t, 6Hz, 2H), 3.13-3.08 (m, 4H), 2.99 (t, 9Hz, 2H), 2.49 (s, 1H), 1.93-1.83 (m, 2H), 1.41 (t, 6Hz, 3H), 1.03 (t, 6Hz, 3H).

[0363] 13 C-NMR (100 MHz, CDCl3): 173.77, 157.58, 153.18, 152.04, 150.20, 149.74, 147.36, 146.66, 145.43, 132.24, 127.40, 127.32, 124.63, 118.73, 114.06, 96.18, 79.37, 74.95, 72.81, 67.93, 66.73, 66.26, 58.25, 49.37, 38.68, 38.78, 37.63, 29.69, 23.19.

[0364] Reaction Scheme 9B (Compound 30B)

[0365] Compound 30B was obtained in 84% yield according to the general synthetic example D using paclitaxel (Pac, 1.20 g, 1.42 mmol) as a drug derivative. The NMR analysis results for compound 30B are as follows.

[0366] 1H-NMR (300 MHz, CDCl3): 8.16 (d, 9Hz, 2H), 7.71 (d, 9Hz, 2H), 7.70-7.28 (m, 10H), 6.98 (d, 9Hz, 1H), 6.31-6.24 (m, 2H), 6.01 (d, 9Hz, 1H), 5.70 (d, 6Hz, 1H), 5.53 (s, 1H), 5.00 (d, 9Hz, 1H), 4.67 (s, 2H), 4.58-4.50 (m, 1H), 4.34 (d, 9Hz, 1H), 4.22 (d, 9Hz, 1H), 3.84 (d, 9Hz, 1H), 2.69-2.35 (m, 13H), 2.23 (s, 3H), 2.03-1.89 (m, 10H), 1.69 (s, 3H), 1.27-1.24 (m, 16H), 1.15 (s, 3H).

[0367] 13 C-NMR (100 MHz, CDCl3): 203.89, 172.19, 172.06, 171.30, 169.89, 168.14, 167.24, 167.05, 142.76, 137.00, 133.73, 133.71, 132.13, 130.31, 129.19, 128.83, 128.60, 127.21, 126.58, 84.52, 79.16, 75.12, 58.57, 52.83, 52.08, 45.67, 43.26, 37.53, 37.11, 32.31, 29.77, 26.88, 24.07, 23.97, 14.91, 14.21, 9.69.

[0368] Reaction Scheme 9C (Compound 30C)

[0369] Compound 30C was obtained in 73% yield according to the general synthetic example D using doxorubicin (Doxo, 0.83 g, 1.42 mmol) as a drug derivative. The NMR analysis results for compound 30C are as follows.

[0370] 1H-NMR (300 MHz, CDCl3): 13.91 (s, 1H), 13.13 (s, 1H), 7.90 (t, 12Hz, 1H), 7.764 (t, 12Hz, 1H), 7.35 (t, 12Hz, 1H), 6.13 (d, 9Hz, 1H), 5.47 (s, 1H), 5.20 (s, 1H), 4.67 (s, 2H), 4.58 (s, 1H), 4.16-4.12 (m, 2H), 4.04 (s, 3H), 3.65 (s, 2H), 3.16 (d, 12Hz, 1H), 2.85 (d, 12Hz, 1H), 2.69-2.65 (m, 4H), 2.47-2.45 (m, 3H), 2.47-2.49 (m, 6H), 1.98-1.92 (m, 2H), 1.48-1.42 (m, 6H).

[0371] 13 C-NMR (100 MHz, CDCl3): 213.97, 186.89, 186.48, 172.33, 171.64, 160.99, 156.24, 155.52, 135.83, 135.37, 133.71, 133.66, 120.68, 119.86, 118.53, 111.49, 111.30, 100.78, 75.13, 69.48, 67.36, 65.60, 56.67, 52.09, 45.37, 37.96, 37.58, 34.62, 32.32, 29.76, 24.71, 24.12, 19.92, 14.19.

[0372] Reaction Scheme 9D (Compound 30D)

[0373] Compound 30D was obtained in 87% yield according to the general synthesis example D using MMAE (1.00 g, 1.42 mmol) as a drug derivative. The NMR analysis results for compound 30D are as follows.

[0374] 1H-NMR (300 MHz, CDCl3): 7.38-7.28 (m, 5H), 6.40 (d, 9Hz, 2H), 4.9 (s, 1H), 4.47 (s, 3H), 4.46-4.50 (m, 2H), 4.43-4.10 (m, 3H), 3.94 (d, 6Hz, 1H), 3.61-3.51 (m, 1H), 3.41 (s, 3H), 3.31 (s, 3H), 3.11-2.94 (m, 5H), 2.80-2.70 (m, 5H), 2.60-2.49 (m, 6H), 2.19-1.98 (m, 10H), 1.31 (m, 6H),1.11-0.07 (m, 26H).

[0375] 13 C-NMR (100 MHz, CDCl3): 174.66, 173.90, 173.25, 172.90, 172.18, 170.65, 170.44, 169.87, 169.79, 141.27, 128.37, 128.10, 127.55, 127.37, 126.39, 126.26, 85.62, 82.05, 75.03, 62.43, 61.01, 60.14, 58.07, 52.05, 51.68, 47.86, 46.62, 44.99, 37.92, 37.57, 32.33, 31.70, 25.07, 24.10, 24.08, 19.42, 18.63, 16.05, 14.50, 13.95, 11.02

[0376] [General Synthesis Example D (Click Synthesis)]

[0377]

[0378] Any one of compounds 30A to 30D is added to compound 28 (P-PEG3-N3, 120 mg, 0.21 mmol, 1.0 equiv), tris[(1-benzyl-1H-1,2,3-triazol-4-yl)methyl]amine (TBTA, 17 mg, 0.032 mmol, 0.15 equiv), and copper(I) iodine (12 mg, 0.063 mmol, 0.30 equiv) in a solvent mixture of H2O / DMF / t-BuOH (0.5 mL / 1.5 mL / 0.5 mL). The reaction mixture is stirred at room temperature under a stream of nitrogen for 5 h. Then, methanol (60 mL) is added, and the precipitated solid is collected by filtration. The filtrate is concentrated under reduced pressure, and the crude product is purified by silica gel column chromatography (MC:MeOH, 95:5 v / v) to obtain the desired P-3C-SS-drug conjugate.

[0379] Synthesis Example D1 (Synthesis of P-3C-SS-SN38): Compound 31A

[0380] Following the general synthesis example D above, compound 30A (138 mg, 0.21 mmol) was selected and used to obtain compound 31A (P-3C-SS-SN38) as a yellow solid (178 mg, yield 81%). The NMR analysis results for compound 31A are as follows.

[0381] 1H-NMR (300 MHz, CDCl3): 9.84-9.74 (m, 1H),8.83 (d, 9Hz, 1H), 8.74-8.73 (m, 1H), 8.25 (d, 9Hz, 1H), 7.80 (s, 1H), 7.72-7.66 (m, 3H),7.57-7.42 (m, 3H), 7.36-7.17 (m, 6H), 6.73 (d, 6Hz, 1H), 6.64 (t, 9Hz, 1H), 5.69 (s, 1H), 5.49 (s, 1H), 5.31-5.19 (m, 7H), 4.66 (d, 6Hz, 1H), 4.45-4.37 (m, 4H), 4.11 (s, 2H), 3.82-3.66 (m, 12H), 3.14-3.12 (m, 2H), 2.84-2.79 (m, 4H), 2.72 (t, 6Hz, 2H), 2.49-2.44 (m, 3H), 2.22-2.17 (m, 3H), 2.06-2.01 (m, 3H), 1.92-1.88 (m, 3H), 1.39 (t, 9Hz, 3H), 1.31 (m, 14H), 1.01 (t, 6Hz, 3H).

[0382] 13 C-NMR (100 MHz, CDCl3): 193.52, 193.05, 173.15, 172.00, 170.73, 167.70, 167.58, 156.93, 149.53, 148.83, 144.62, 140.36, 135.99, 131.36, 128.37, 124.066, 124.16, 119.29, 113.89, 110.71, 97.39, 83.34, 72.11, 69.77, 65.60, 56.91, 53.48, 49.49, 45.27, 36.92, 37.74, 29.00, 24.18, 23.42, 23.22, 13.43, 7.12.

[0383] 합성예 D2 (P-3C-SS-Pac의 합성): 화합물 31B

[0384] Following the general synthesis example D above, compound 30B (233 mg, 0.21 mmol) was selected and used to obtain compound 31B (P-3C-SS-Pac) as a white solid (291 mg, yield 93%). The NMR analysis results for compound 31B are as follows.

[0385] 1 H-NMR (300 MHz, CDCl3): 9.88 (m, 1H), 8.82-8.77 (m, 2H), 8.18-8.13 (m, 2H), 7.78 (d, 9Hz, 2H), 7.68-7.33 (m, 16H), 7.10 (d, 9Hz, 1H), 6.78-6.52 (m, 2H), 6.32-6.25 (m, 2H), 6.02-5.98 (m, 1H), 5.70 (d, 6Hz, 1H), 5.54 (d, 6Hz, 1H), 5.32 (s, 1H), 5.18 (s, 2H), 5.01 (d, 9Hz, 1H), 4.70 (d, 6Hz, 1H), 4.49-4.33 (m, 4H), 4.36 (d, 9Hz, 1H), 4.21 (d, 3Hz, 1H), 4.11 (s, 2H), 3.85-3.66 (m, 12H), 2.67-2.41 (m, 16H), 2.25 (s, 3H), 2.20-2.19 (m, 1H), 2.03-1.87 (m, 10H), 1.74-1.71 (m, 6H), 1.34-1.25 (m, 17H), 3.66 (s, 3H).

[0386] 13C-NMR (100 MHz, CDCl3): 214.08, 193.84, 181.07, 186.66, 172.85, 171.60, 168.48, 161.06, 155.69, 150.71, 142.56, 141.19, 136.73, 135.76, 128.84, 128.04, 125.10, 122.98, 118.48, 114.93, 111.57, 111.47, 100.96, 84.12, 83.68, 70.57, 70.53, 70.33, 69.27, 57.49, 56.72, 50.32, 46.04, 45.33, 38.17, 37.81, 32.49, 29.77, 24.94, 24.73, 24.28, 17.01, 14.20.

[0387] Synthesis example D3 (synthesis of P-3C-SS-Doxo): Compound 31C

[0388] Following the general synthesis example D above, compound 30C (165 mg, 0.21 mmol) was selected and used to obtain compound 31C (P-3C-SS-Doxo) as a red solid (228 mg, 78% yield). The NMR analysis results for compound 31C are as follows.

[0389] 1H-NMR (300 MHz, CDCl3): 13.96 (s, 1H),13.22 (s, 1H),9.80 (s, 1H), 8.83 (s, 1H),8.74 (t, 6Hz, 1H), 8.01(s, 1H), 7.90 (d, 6Hz, 1H), 7.74-7.71(m, 3H),7.46-7.29 (m, 5H),6.75 (d, 9Hz, 1H), 6.66 (t, 9Hz, 1H),6.65 (S, 1H),5.50 (s, 1H),5.26 (s, 1H),5.19(s, 2H),4.77 (s, 2H), 4.69 (s, 3H),4.41(t, 6Hz, 2H), 4.16-4.07 (m, 7H), 3.81-3.67 (m, 11H),3.27-2.94 (m, 4H), 2.64 (t, 6Hz, 4H), 2.46-2.41(m, 3H),2.33-2.19 (m, 4H), 2.01-1.84 (m, 8H), 1.33-1.21 (m, 40H), 0.90-0.84 (m, 3H)

[0390] 13 C-NMR (100 MHz, CDCl3): 214.08, 193.84, 187.07, 186.66, 172.85, 171.60, 168.48, 161.06, 155.69, 150.71, 147.56, 141.19, 136.73, 135.76, 128.84, 128.04, 125.10, 122.98, 118.48, 114.93, 111.57, 111.47, 100.96, 84.12, 70.57, 70.53, 70.33, 69.27, 57.49, 56.72, 50.32, 46.04, 38.17, 37.81, 29.77, 24.94, 24.73, 24.28, 17.09, 14.20

[0391] 합성예 D4 (P-3C-SS-MMAE의 합성): 화합물 31D

[0392] Following the general synthesis example D above, compound 30D (198 mg, 0.21 mmol) is selected and used to obtain compound 31D (P-3C-SS-MMAE) as a pale yellow solid (274 mg, yield 89%). The NMR analysis results for compound 31D are as follows.

[0393] 1 H-NMR (300 MHz, CDCl3): 9.81 (s, 1H), 8.85-8.76 (m, 2H), 8.14 (s, 1H), 7.92-7.73 (m, 2H), 7.73-7.44 (m, 3H), 7.31-7.20 (m, 8H), 6.76-6.51 (m, 3H), 5.19 (s, 2H), 4.95 (s, 1H), 4.73-4.57 (m, 3H), 4.46-4.37 (m, 3H), 4.29-4.27 (m, 1H), 4.22-4.12 (m, 3H), 4.04-4.02 (m, 1H), 3.83-3.66 (m, 11H), 3.55-3.47 (m, 1H), 3.41 (s, 3H), 3.37 (s, 3H), 3.03-2.98 (m, 5H), 2.75-2.66 (m, 4H), 2.50-2.35 (m, 7H), 2.24-2.23 (m, 3H), 2.02-1.98 (m, 9H), 1.83-1.80 (m, 3H), 1.33-1.25 (m, 12H), 1.04-0.81 (m, 29H).

[0394] 13C-NMR (100 MHz, CDCl3): 198.99, 198.81, 174.79, 173.30, 172.89, 172.78, 170.64, 169.85, 168.49, 168.36, 150.73, 150.23, 142.63, 141.14, 136.77, 136.00, 128.11, 126.77, 124.91, 120.10, 115.78, 111.50, 97.95, 85.66, 84.10, 70.55, 70.51, 60.13, 58.00, 57.66, 54.09, 50.26, 47.89, 46.04, 37.87, 37.62, 32.49, 29.76, 24.94, 24.11, 19.43, 18.64, 14.19, 11.00.

[0395] Reaction Scheme 10 (Compound 32)

[0396]

[0397] Bis-PEG4 (1.70 g, 3.60 mmol), EDC·HCl (0.70 g, 5.20 mmol), and DMAP (110 mg, 0.90 mmol) are dissolved in anhydrous dichloromethane (10 mL) under a nitrogen stream at 0°C. The reaction mixture is stirred for 30 min, and then compound 7 (P-NH2, 0.50 g, 1.85 mmol) is added. The mixture is heated to room temperature and stirred vigorously for 12 h. The reaction progress is monitored by TLC (silica gel, confirmed by a UV lamp). After completion of the above process, the solvent is removed under reduced pressure, and the residue is purified by silica gel column chromatography (methylene chloride / methanol, 95:5 v / v) to give compound 32 (P-PEG4) as a colorless solid (1.15 g, 51% yield). The NMR analysis results for compound 32 are as follows.

[0398] 1H-NMR (300 MHz, CDCl3): 9.81 (s, 1H), 8.74 (s, 1H), 7.84 (s, 1H), 7.84 (d, 9Hz, 1H), 7.46 (d, 9Hz, 1H), 7.44-7.34 (m, 2H), 6.76 (d, 9Hz, 1H), 6.63 (t, 6Hz, 1H), 4.68 (s, 2H), 3.86-3.65 (m, 20H), 2.68-2.49 (m, 4H), 1.37 (s, 12H).

[0399] [General synthesis example E (synthesis of Ac-SL-D): SL = PEG4]

[0400]

[0401] The reactions mentioned in General Synthesis Example D are carried out under a nitrogen stream at room temperature (rt) unless otherwise specified. The progress of the reaction is monitored by chromatography (TLC) on silica gel plates (confirmed by ultraviolet light or appropriate staining), and column chromatography is performed on silica gel (230-400 mesh) using methylene chloride / methanol (MC / MeOH) as the eluent. To a solution of compound 32 (P-PEG4) in anhydrous DCM (5 mL) are added DMAP and EDC·HCl in the amounts specified below. The mixture is stirred at rt for 30 min under nitrogen. Thereafter, each anticancer drug (SN38, paclitaxel, doxorubicin, monomethyl auristatin E) is added, and the reaction mixture is stirred vigorously at rt for 12 h. After completing the above process, monitor by TLC, evaporate the solvent under reduced pressure, and purify the product by silica gel column chromatography (MC / MeOH, 95:5, v / v) to obtain the desired complex.

[0402] Synthesis Example E1 (Synthesis of P-PEG4-SN38): Compound 33A

[0403] Compound 32 (P-PEG4, 0.15 g, 0.283 mmol), DMAP (0.05 g, 0.40 mmol), and EDC·HCl (0.077 g, 0.40 mmol) are dissolved in DCM (5 mL). After stirring for 30 min, SN38 (0.078 g, 0.20 mmol) is added, and the reaction mixture is stirred at room temperature for 12 h. As a result of purification, compound 33A (0.18 g, 61% yield) is obtained as a pale yellow solid. The NMR analysis results for compound 33A are as follows.

[0404] 1 H-NMR (300 MHz, CDCl3): 9.83 (s, 1H), 8.26 (s, 1H), 8.54 (s, 1H), 8.26 (d, 9Hz, 1H), 7.84-7.80 (m, 3H), 7.66 (s, 1H), 7.59 (d, 9Hz, 1H), 7.34 (d, 9Hz, 1H), 7.34-7.31 (m, 2H), 6.67 (d, 9Hz, 1H), 6.52 (t, 9Hz, 6H), 5.71 (d, 15Hz, 1H), 5.31 (d, 15Hz, 1H), 5.30 (s, 1H), 4.67-4.59 (m, 2H), 3.91 (t, 2Hz, 2H), 3.81-3.96 (m, 16H), 3.31-3.29 (m, 2H), 2.91 (t, 6Hz, 3H), 2.72-2.61 (m, 2H), 1.98-1.89 (m, 2H), 1.71-1.61 (m, 8H), 1.49-1.21 (m, 18H) 1.07 (t,3Hz, 6H).

[0405] Synthesis Example E2 (Synthesis of P-PEG4-Pac): Compound 33B

[0406] Compound 32 (P-PEG4, 0.15 g, 0.283 mmol), DMAP (0.10 g, 0.80 mmol), and EDC·HCl (0.90 g, 2.14 mmol) are dissolved in DCM (5 mL). After stirring for 30 min, paclitaxel (Pac, 0.170 g, 0.20 mmol) is added, and the reaction mixture is stirred at room temperature for 12 h. As a result of purification, compound 33B (0.18 g, 63% yield) is obtained as a white solid. The NMR analysis results for compound 33B are as follows.

[0407] 1 H-NMR (300 MHz, CDCl3): 9.79 (s, 1H), 8.81 (t, 6Hz, 1H), 8.63 (s, 1H), 8.16 (d, 9Hz, 2H), 7.92-7.81 (m, 4H), 7.61-7.21 (m, 18H), 6.81 (d, 9Hz, 1H), 6.72 (t, 6Hz, 1H), 6.39 (m, 1H), 6.31 (t, 6Hz, 1H), 5.99 (m, 1H), 5.78 (d, 9Hz, 1H), 5.71 (d, 9Hz, 1H), 4.91 (d, 9Hz,1H), 4.79 (d, 9Hz, 2H), 4.51 (s, 1H), 4.31 (d, 9Hz,1H), 4.21 (d, 9Hz,1H), 3.89-3.39 (m, 22H), 2.78-2.51 (m, 8H), 2.49 (s, 3H), 2.41-2.19 (m, 13H), 1.99-1.89 (m, 10H) 1.71 (s, 3H), 1.37-1.28 (m, 22H)

[0408] Synthesis Example E3 (Synthesis of P-PEG4-Doxo): Compound 33C

[0409] Compound 32 (P-PEG4, 0.15 g, 0.283 mmol), DMAP (0.10 g, 0.10 mmol), and EDC·HCl (0.90 g, 2.14 mmol) are dissolved in DCM (5 mL). After stirring for 30 min, doxorubicin·HCl (Doxo, 0.115 g, 0.20 mmol) is added, and the reaction mixture is stirred at room temperature for 12 h. As a result of purification, compound 33C (0.10 g, 43% yield) is obtained as a red solid. The NMR analysis results for compound 33C are as follows.

[0410] 1 H-NMR (300 MHz, CDCl3): 13.95 (s, 1H), 13.23 (s, 1H), 9.81 (s, 1H), 8.77 (s, 1H), 8.73 (s, 1H), 8.03 (d, 9Hz, 1H), 7.82-7.63 (m, 3H), 7.46-7.29 (m, 4H), 6.87 (d, 9Hz, 1H), 6.74 (d, 6Hz, 1H), 6.65 (t, 9Hz, 1H), 5.51 (s, 1H), 5.57 (s, 1H), 4.77-4.65 (m, 4H), 4.41-4.39 (m, 1H), 4.13-4.07 (m, 4H), 3.84-3.59 (m, 42H), 3.28 (m, 1H), 3.01-2.89 (m, 3H), 2.63 (t, 6Hz, 3H), 2.48-2.29 (m, 5H), 2.18-2.17 (m, 1H), 1.91-1.69 (m, 1H), 1.79-1.75 (m, 1H), 1.33-1.26 (m, 14H), 0.89-0.86 (m, 3H).

[0411] Synthesis Example E4 (Synthesis of P-PEG4-MMAE): Compound 33D

[0412] Compound 32 (P-PEG4, 0.15 g, 0.283 mmol), DMAP (0.10 g, 0.10 mmol), and EDC·HCl (0.90 g, 2.14 mmol) are dissolved in DCM (5 mL). After stirring for 30 min, monomethyl auristatin E (MMAE, 0.143 g, 0.20 mmol) is added, and the reaction mixture is stirred at room temperature for 12 h. As a result of purification, compound 33D (0.21 g, 67% yield) is obtained as a white solid. The NMR analysis results for compound 33D are as follows.

[0413] 1 H-NMR (300 MHz, CDCl3): 9.77 (s, 1H), 8.76 (t, 6Hz, 1H), 8.59 (s, 1H), 7.81-7.75 (m, 2H), 7.45 (d, 6Hz, 1H), 7.39-7.24 (m, 6H), 6.74 (d, 6Hz, 1H), 6.67-6.55 (m, 2H), 4.68 (d, 9Hz, 2H), 4.62-4.56 (m, 1H), 4.28-4.06 (m, 3H), 3.83-3.76 (m, 6H), 3.67-3.55 (m, 30H), 3.40 (s, 3H), 3.31 (s, 3H), 3.00-2.97 (m, 4H), 2.67-2.61 (m, 4H), 2.03-1.96 (m, 2H), 1.84-1.82 (m, 2H), 1.32 (s, 12H), 1.26 (s, 6H), 1.04-0.81(m, 12H).

Claims

1. Albumin-binding prodrug represented by the following chemical formula I: <Chemical Formula I> Ac-(CL1) c1 -(SL1) s1 -(CL2) c2 -(SL2) s2 -D In the above chemical formula I, Ac is an albumin-binding moiety represented by the following chemical formula 1, CL1 and CL2 are each a cleavable linker, SL1 and SL2 are each soluble linkers, c1, c2, s1 and s2 are each 0 or 1, D is a drug moiety, <Chemical Formula 1> In the above chemical formula 1, Z is N, O or C(R 14 ) and, R 11 Inland R 14 , R 21 Inland R 24 And R3 to R6 are each independently hydrogen, deuterium, -F, -Cl, -Br, -I, hydroxyl group, cyano group, nitro group, formyl group, carboxyl group, amino group, substituted or unsubstituted C1-C 10 Alkyl group, substituted or unsubstituted C2-C 10 Alkenyl group, substituted or unsubstituted C2-C 10 Alkynyl group, substituted or unsubstituted C1-C 10 Alkoxy group, substituted or unsubstituted C6-C 10 Aryl group, -N(Q1)(Q2) and -C(=O)(Q1), selected from R 11 Inland R 14 and R 21 Inland R 24 One of them is (CL1) in the above chemical formula I c1 -(SL1) s1 -(CL2) c2 -(SL2) s2 -This is a site that combines with D, Optionally i) R 11 and R 12 , ii) R 12 Wow R 13 , iii) R 13 and R 14 , iv) R 21 and R 22 , v) R 22 Wow R 23 , vi) R 23 and R 24 , or vii) any combination thereof, which may be combined with each other to form a substituted or unsubstituted C3-C 10 Carbocyclic group or substituted or unsubstituted C1-C 10 Forming a heterocyclic group, Q1 and Q2 are independently hydrogen, deuterium, -F, -Cl, -Br, -I, hydroxyl group, cyano group, nitro group, formyl group, carboxyl group, amino group, substituted or unsubstituted C1-C 10 Alkyl group, substituted or unsubstituted C2-C 10 Alkenyl group, substituted or unsubstituted C2-C 10 Alkynyl group, substituted or unsubstituted C1-C 10 Alkoxy group and substituted or unsubstituted C6-C 10 Selected from aryl groups, substituted C1-C 10 Alkyl group, substituted C2-C 10 Alkenyl group, substituted C2-C 10 alkynyl group, substituted C1-C 10 Alkoxy group, substituted C6-C 10 Aryl group, substituted C3-C 10 Carbocyclic groups and substituted C1-C 10 The substituents of the heterocyclic group are deuterium, -F, -Cl, -Br, -I, hydroxyl group, cyano group, nitro group, formyl group, carboxyl group, amino group, C1-C 10 Alkyl group and C6-C 10 Selected from aryl groups, m is an integer selected from 1 to 5.

2. In paragraph 1, An albumin-binding prodrug, wherein Ac in the above chemical formula I is an albumin-binding moiety represented by any one of the following chemical formulas 1-1 to 1-6: <Chemical Formula 1-1> <Chemical Formula 1-2> <Chemical Formula 1-3> <Chemical Formula 1-4> <Chemical Formula 1-5> <Chemical Formula 1-6> Among the above chemical formulas 1-1 to 1-6, Z, R 11 Inland R 13 , R 21 Inland R 24 , R3 to R6 and Q1 are each the same as described in the above chemical formula 1, *is (CL1) in the above chemical formula I c1 -(SL1) s1 -(CL2) c2 -(SL2) s2 -This is a site that is combined with D.

3. In paragraph 1, In the above chemical formula I, D is Camptothecin, paclitaxel, doxorubicin, auristatin E, auristatin F, monomethyl auristatin E (MMAE), monomethyl auristatin F (MMAF), cyclophosphamide, mechlorethamine, uramustine, melphalan, chlorambucil, ifosfamide, bendamustine, carmustine, lomustine, streptozocin, busulfan, dacarbazine, temozolomide, thiotepa, altretamine, duocarmycin, cisplatin, carboplatin, nedaplatin, oxaliplatin, satraplatin, triplatin tetranitrate, 5-fluorouracil, 6-mercaptopurine, capecitabine, cladribine, clofarabine, cystabine, floxuridine, fludarabine, gemcitabine, hydroxyurea, methotrexate, pemetrexed, pentostatin, thioguanine, topotecan, exatecan,An albumin-binding prodrug characterized by at least one selected from the group consisting of irinotecan, etoposide, teniposide, mitoxantrone, docetaxel, izabepilone, vinblastine, vincristine, vindesine, vinorelbine, estramustine, maytansine, mertansine, dolastatin, and derivatives thereof.

4. In paragraph 1, The above albumin-binding prodrug is an albumin-binding prodrug characterized in that it binds to albumin present in the blood through injection administration to form an albumin-drug conjugate.

5. A pharmaceutical composition for preventing or treating cancer, comprising an albumin-binding prodrug according to Article 1 as an active ingredient.

6. In paragraph 5, The above cancers include stomach cancer, lung cancer, non-small cell lung cancer, breast cancer, kidney cancer, ovarian cancer, liver cancer, bronchial cancer, nasopharyngeal cancer, oral cancer, laryngeal cancer, pancreatic cancer, bladder cancer, colon cancer, cervical cancer, bone cancer, non-small cell bone cancer, blood cancer, skin cancer, head or neck cancer, uterine cancer, rectal cancer, colon cancer, anal cancer, fallopian tube cancer, endometrial cancer, vaginal cancer, fallopian tube cancer, vulvar cancer, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, chronic or acute leukemia, lymphocytic lymphoma, kidney or ureter cancer, renal cell carcinoma, renal pelvic carcinoma, salivary gland cancer, sarcoma, pseudomyxoma, hepatoblastoma, testicular cancer, glioblastoma, lip cancer, ovarian germ cell tumor, A pharmaceutical composition for preventing or treating cancer, wherein the pharmaceutical composition is at least one selected from the group consisting of basal cell carcinoma, multiple myeloma, gallbladder cancer, choroidal melanoma, ampulla of Vater cancer, peritoneal cancer, tongue cancer, small cell carcinoma, pediatric lymphoma, neuroblastoma, duodenal cancer, ureteral cancer, astrocytoma, meningioma, renal pelvis cancer, vulvar cancer, thymic cancer, central nervous system tumor, primary central nervous system lymphoma, spinal cord tumor, brainstem glioma, and pituitary adenoma.

7. A method for producing an albumin-drug conjugate, comprising a step of reacting an albumin-binding prodrug according to paragraph 1 with albumin.

8. Albumin-drug conjugate represented by the following chemical formula II: <Chemical Formula II> Ab-(CL1) c1 -(SL1) s1 -(CL2) c2 -(SL2) s2 -D In the above chemical formula II, Ab is an albumin binding moiety represented by the following chemical formula 2, CL1 and CL2 are each a cleavable linker, SL1 and SL2 are each soluble linkers, c1, c2, s1 and s2 are each 0 or 1, D is a drug moiety, <Chemical Formula 2> In the above chemical formula 2, A is a group excluding amine groups in albumin, Z is N, O or C(R 14 ) and, R 11 Inland R 14 , R 21 Inland R 24 And R3 to R6 are each independently hydrogen, deuterium, -F, -Cl, -Br, -I, hydroxyl group, cyano group, nitro group, formyl group, carboxyl group, amino group, substituted or unsubstituted C1-C 10 Alkyl group, substituted or unsubstituted C2-C 10 Alkenyl group, substituted or unsubstituted C2-C 10 Alkynyl group, substituted or unsubstituted C1-C 10 Alkoxy group, substituted or unsubstituted C6-C 10 Aryl group, -N(Q1)(Q2) and -C(=O)(Q1), selected from R 11 Inland R 14 and R 21 Inland R 24 One of them is (CL1) in the above chemical formula II c1 -(SL1) s1 -(CL2) c2 -(SL2) s2 -This is a site that combines with D, Optionally i) R 11 and R 12 , ii) R 12 Wow R 13 , iii) R 13 and R 14 , iv) R 21 and R 22 , v) R 22 Wow R 23 , vi) R 23 and R 24 , or vii) any combination thereof, which may be combined with each other to form a substituted or unsubstituted C3-C 10 Carbocyclic group or substituted or unsubstituted C1-C 10 Forming a heterocyclic group, Q1 and Q2 are independently hydrogen, deuterium, -F, -Cl, -Br, -I, hydroxyl group, cyano group, nitro group, formyl group, carboxyl group, amino group, substituted or unsubstituted C1-C 10 Alkyl group, substituted or unsubstituted C2-C 10 Alkenyl group, substituted or unsubstituted C2-C 10 Alkynyl group, substituted or unsubstituted C1-C 10 Alkoxy group and substituted or unsubstituted C6-C 10 Selected from aryl groups, substituted C1-C 10 Alkyl group, substituted C2-C 10 Alkenyl group, substituted C2-C 10 alkynyl group, substituted C1-C 10 Alkoxy group, substituted C6-C 10 Aryl group, substituted C3-C 10 Carbocyclic groups and substituted C1-C 10 The substituents of the heterocyclic group are deuterium, -F, -Cl, -Br, -I, hydroxyl group, cyano group, nitro group, formyl group, carboxyl group, amino group, C1-C 10 Alkyl group and C6-C 10 Selected from aryl groups, m is an integer selected from 1 to 5.

9. A pharmaceutical composition for preventing or treating cancer, comprising an albumin-drug conjugate according to Article 8 as an active ingredient.

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