WO2023104021A1

WO2023104021A1 - Metabolic checkpoint-based human serum albumin nano-drug, and preparation method therefor and application thereof

Info

Publication number: WO2023104021A1
Application number: PCT/CN2022/136879
Authority: WO
Inventors: 蔡林涛; 黄国俊; 潘宏; 郑明彬; 唐晓帆; 廖健洪; 王盟盟; 张保珍
Original assignee: 深圳先进技术研究院
Priority date: 2021-12-07
Filing date: 2022-12-06
Publication date: 2023-06-15
Also published as: CN114306620A

Abstract

A metabolic checkpoint-based human serum albumin nano-drug (IDNPs), and a preparation method therefor and an application thereof. A prodrug molecule IDOi-DON obtained by coupling an indoleamine-2,3-dioxygenase inhibitor (IDOi) and a metabolic reprogramming agent DON by means of a chemical bond amidation reaction can be subjected to enzymatic cleavage in a tumor microenvironment, and the outer surface is coated with natural biocompatible serum albumin. The drug stability is improved, the toxic and side effects of the drug are reduced, the drug is enriched at a tumor site, the effects of inhibiting the IDO activity of tumor cells and inhibiting Treg cell proliferation while activating CD8+T cells are achieved by a synergistic effect of two drugs, and the anti-tumor immunotherapy effect is synergistically enhanced.

Description

Human serum albumin nanomedicine based on metabolic checkpoint and its preparation method and application

technical field

The invention relates to the field of biomedical technology, in particular to a human serum albumin nano-medicine based on a metabolic checkpoint and its preparation method and application.

Background technique

Tumor immunotherapy is a revolutionary breakthrough in the field of tumors in recent years by stimulating the body's anti-tumor immune response to kill or inhibit tumor growth. However, there is still an important problem that the immune tolerance formed by the tumor microenvironment cannot be overcome. The immune microenvironment caused by the vigorous metabolism of the tumor forms a metabolic inhibition of immune cells, and cannot effectively recognize and kill tumor cells. Therefore, for the metabolic targets of tumor immunosuppression, it is of great significance to develop functional metabolic check regulators that induce controllable immune responses for cancer immunotherapy.

Tumor immunotherapy inhibits tumor progression by activating immune cells to identify and eliminate tumor cells. The tumor immunosuppressive microenvironment is a key factor leading to tumor growth and metastasis. A variety of immunosuppressive cells in the tumor microenvironment can inhibit the anti-tumor immune response, such as interleukin-10 (IL-10), indoleamine 2, 3-dioxygenase (IDO), etc. Immunosuppressive factors in the tumor microenvironment severely inhibit the activation, migration and differentiation of immune cells, limiting the responsiveness of anti-tumor immunotherapy. IDO inhibitors such as 1-MT and NLG919 can inhibit the degradation of tryptophan, reverse the inhibition of dendritic cells on T cells, activate T cells to generate immune responses, and finally suppress or eliminate tumors. However, due to the complexity of the immunosuppressive microenvironment, it is difficult to reactivate T cells only by inhibiting one immunosuppressive pathway. IDOi has shown weak anti-tumor effects and obvious drug side effects in clinical studies. Therefore, there is an urgent need to design agents or molecules that simultaneously target multiple targets to improve the efficacy of immunotherapy.

Contents of the invention

Aiming at the defects in the prior art, the present invention proposes a human serum albumin nano-medicine based on a metabolic checkpoint and its preparation method and application. Coupling indoleamine-2,3-dioxygenase inhibitor (IDOi) and metabolic reprogramming agent DON by chemical bond amidation reaction to obtain prodrug molecule IDOi-DON, and then coated with natural biocompatible human serum white protein to obtain the human serum albumin nanomedicine based on metabolic checkpoints.

The invention provides a human serum albumin nano-medicine based on a metabolic checkpoint, comprising a prodrug molecular core and human serum albumin coated on the outside, the prodrug molecule being indoleamine-2,3-dioxygenase A coupling molecule of an inhibitor and a metabolic reprogramming agent, the coupling mode is amide bond coupling.

Further, the metabolic reprogramming agent is a glutamine antagonist.

Further, the metabolic reprogramming agent is selected from 6-diazo-5-oxo-norleucine, 5-diazo-4-oxo-L-norvaline, L-(α-S, Any one of 5S)-α-amino-3-chloro-4,5-dihydro-5-isoxazoleacetic acid and azaserine has the same connection principle.

Further, the indoleamine-2,3-dioxygenase inhibitor is an immune activator.

Further, the immune activator is 1-methyl tryptophan.

Further, the human serum albumin coats the inner core of the prodrug molecule through hydrophobic interaction.

Further, the structure of the prodrug molecular core is shown in formula (I):

Wherein, n is any one of 0, 1, 2, 3, 4, 5, 6, 7; R ₁ and R ₂ are selected from H or alkyl, substituted alkyl, cycloalkyl, substituted cycloalkane radical, aryl, _substituted aryl, heterocyclic, substituted heterocyclic, alkenyl, substituted alkenyl, cycloalkenyl, substituted cycloalkenyl, heteroaryl, substituted heteroaryl; is H or is capable of forming an amide bond, a carbamate bond, a phosphoramidate bond, a phosphoramidate bond with the nitrogen adjacent to _R3 ; _R3 ' is selected from H or C1-C8 alkyl, substituted C1- C8 alkyl, or R ₃ and R ₃ ' together form a ring structure comprising -C(=O)-GC(=O)-, wherein G is selected from the group consisting of: C1-C8 alkylene, C1-C8 Heteroalkylene, C5-C8 cycloalkylene, C6-C12 arylene, C5-C12 heteroarylene, divalent C4-C10 heterocycle, each of which may be optionally substituted.

The present invention also provides the preparation method of the human serum albumin nano-medicine based on the metabolic checkpoint, comprising the following steps:

(1) Synthesis of 9-fluorenylmethoxycarbonyl-protected (indoleamine-2,3-dioxygenase inhibitor)-metabolic reprogramming agent;

(2) Preparation of (indoleamine-2,3-dioxygenase inhibitor)-metabolic reprogramming agent prodrug molecule by removing the carbonyl group of 9-fluorenylmethoxy;

(3) Preparation of (indoleamine-2,3-dioxygenase inhibitor)-metabolic reprogramming agent human serum albumin nanomedicine.

Further, in the step (1), the isopropyl-metabolic reprogramming agent and 9-fluorenylmethoxycarbonyl-indoleamine-2,3-dioxygenase inhibitor are combined in benzotriazole-N ,N,N',N'-Tetramethyluronium hexafluorophosphate and diisopropylethylamine were coupled to generate 9-fluorenylmethoxycarbonyl-protected indoleamine-2,3-bis Oxygenase inhibitors - metabolic reprogramming agents.

Further, in the step (2), the product obtained in the step (1) is reacted overnight under the action of piperidine to remove the carbonyl group of the 9-fluorenylmethoxy group to obtain the prodrug molecule.

Further, in the step (3), (1) dissolve the prodrug molecule obtained in step (2) in an organic solvent, and slowly drop the prodrug molecule solution into the human serum albumin under the continuous action of external force. In the aqueous solution, self-assembly of human serum albumin molecules is induced, purified and filtered to obtain the human serum albumin nano-medicine based on metabolic checkpoints.

The present invention also provides the application of the human serum albumin nano-medicine based on metabolic checkpoint in tumor immunotherapy.

In summary, compared with the prior art, the present invention achieves the following technical effects:

(1) The present invention prepares a prodrug molecule by chemically coupling IDOi and a metabolic reprogramming agent, which can be enzymatically broken into two drug molecules in the tumor microenvironment, and synergistically act on tumor cells in time and space to inhibit tumor growth.

(2) The present invention uses the self-assembly technology of human serum albumin molecules induced by hydrophobic drugs to prepare albumin nano-drugs (IDNPs) of IDOi-DON, which can improve the stability of the drug, reduce the toxic and side effects of the drug, and increase the enrichment of tumor sites , improve the efficiency of tumor treatment, and have good biological safety, and overcome the toxicity challenges faced by IDOi and DON in clinical application.

(3) IDOi of the present invention inhibits the proliferation of Treg cells and DON activates CD8 ⁺ T cells to synergistically enhance anti-tumor immune response and effectively improve the effect of immunotherapy.

Description of drawings

In order to illustrate the technical solutions of the embodiments of the present invention more clearly, the accompanying drawings used in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of the present invention, and thus It should be regarded as a limitation on the scope, and those skilled in the art can also obtain other related drawings based on these drawings without creative work.

Figure 1 is the H NMR spectrum of IDOi-DON.

Figure 2 is the mass spectrum of IDOi-DON.

Figure 3 shows the particle size distribution and transmission electron microscope images of IDNPs.

Figure 4 is the growth inhibition evaluation of IDNPs on SPCA1 lung cancer cells.

Detailed ways

In order to enable those skilled in the art to better understand the solutions of the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only It is an embodiment of a part of the present invention, but not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

Indoleamine-2,3-dioxygenase (Indoleamine-2,3-dioxygenase, IDO)-mediated tryptophan metabolite is an important physiological substance for anti-tumor immunosuppression, so IDO is an important component of tumor immunotherapy. Metabolic targets (metabolic checkpoints). IDO has the ability to catalyze the degradation of tryptophan. The metabolite kynurenine mediated by IDO can not only promote angiogenesis, but also inhibit the function of immune cells, thereby escaping the body's ability to monitor and eliminate tumor cells. IDO is highly expressed in tumors and metastatic lymph nodes, and is involved in the immune tolerance mechanism of tumors. IDO inhibitors (IDO inhibitors, IDOi) represented by 1-methyltryptophan (1-MT) can inhibit the growth of tumor cells by enhancing the anti-tumor immune response. However, in preclinical studies, IDOi only showed moderate anti-tumor activity in vivo, and combined administration is limited by complex pharmacokinetics and drug-drug interactions, so it is necessary to design multiple metabolic checkpoints. A molecule that overcomes these inherently challenging issues.

Metabolic reprogramming is also a hallmark of malignant tumors. Cancer progression depends on metabolic reprogramming. Inhibiting certain metabolic pathways of tumors is also an important method for cancer treatment. Glutamine metabolism is also closely related to tumor growth and survival. Glutamine generates glutamic acid under the action of glutaminase, and then glutamic acid is converted into α-ketoglutarate and enters tricarboxylic acid (tricarboxylic acid, TCA) cycle plays an important role in anabolism. A structural analogue of glutamine, 6-diazo-5-oxo-L-norleucine (DON), can very broad-spectrum and effectively inhibit the activity of enzymes that use glutamine as a substrate, and DON inhibits Disabling glutamine metabolism will inhibit the oxidative phosphorylation of tumor cells, and at the same time promote the proliferation and activation of CD8 ⁺ T cells, and then cooperate with IDOi to ensure a strong and sustained immune response. However, the oxidative phosphorylation metabolic pathway is elevated in T cells, which can activate CD8 ⁺ T cells and enhance anti-tumor immunotherapy. However, due to its high toxicity, DON was terminated in clinical phase II trials and its application was limited. Simple drug combination will face complex pharmacokinetic problems, and drug side effects cannot be avoided. Therefore, chemically coupled IDOi and DON can target two immune metabolic pathways, inhibit tumor cell metabolism and enhance T cell activity. , a feasible method to weaken the side effects of drugs, and is expected to comprehensively improve the inhibition of tumor growth.

Nanoparticles coated with human serum albumin (HSA) can increase the bioavailability of drugs, and take advantage of the biocompatibility of HSA and the high permeability and retention (Enhanced Permeability and Retention, EPR) effect of solid tumors The passive targeting of nanomedicines increases the enrichment of nanomedicines at tumor sites and can overcome the challenges of clinical toxicity.

The rapid development of nanotechnology has made a leap in the development of targeted therapy and in vivo diagnosis of tumors. As a drug carrier, nanomaterials can improve the stability of drugs, reduce the toxic and side effects of drugs, enable precise release of drugs at tumor sites or achieve sustained and controlled release effects, and overcome the shortcomings of traditional drugs such as low bioavailability and serious adverse reactions.

Based on the above reasons, the present invention designs a protein nano-drug that encapsulates a prodrug of a metabolic reprogramming agent-immune activator. -DON prodrug molecule. The prodrug molecule IDOi-DON is obtained through amidation coupling and deprotection of IDOi and DON. The prodrug molecule prepared by chemically coupling IDOi and the metabolic reprogramming agent DON can be cleaved in response to enzymes in the tumor microenvironment and coated with natural biocompatible serum albumin. This functional protein nano-drug can be enriched in the tumor site, Inhibit the oxidative phosphorylation of tumor cells, inhibit the proliferation of Treg cells, activate CD8 ⁺ T cells, and enhance the effect of anti-tumor immunotherapy.

The nanometer prodrug based on the metabolic checkpoint of the present invention is a double-drug coupling prodrug molecule, which is a metabolic reprogramming agent-immune activator coupled prodrug molecule (IDOi-DON).

The prodrug molecule (IDOi-DON) coupled with metabolic reprogramming agent-immune activator has a structure of formula (I).

The metabolic reprogramming agent is a glutamine antagonist.

The metabolic reprogramming agent is selected from 6-diazo-5-oxo-norleucine (DON), 5-diazo-4-oxo-L-norvaline (L-DONV), assi Any one of acivicin (L-(α-S,5S)-α-amino-3-chloro-4,5-dihydro-5-isoxazoleacetic acid) and azaserine.

The immunoactivator is indoleamine-2,3-dioxygenase inhibitor (IDOi) 1-methyltryptophan.

The preparation method of the double drug coupling prodrug molecule comprises: Fmoc group protected 1-methyl tryptophan (1-MT) and isopropyl-DON are coupled through an amide bond, and the obtained coupling The molecule is detached from the Fmoc group through a hydrolysis reaction to obtain a double-drug coupling prodrug molecule, which specifically includes the following steps:

(1) Isopropyl-DON and 9-fluorenylmethoxycarbonyl-1-methyltryptophan in benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate In the presence of (HBTU) and diisopropylethylamine, coupled to generate Fmoc-protected IDOi-DON;

(2) The Fmoc-protected IDOi-DON obtained in step (1) reacted overnight under the action of piperidine to remove the Fmoc protecting group to obtain the target product IDOi-DON;

The present invention also provides a method for preparing human serum albumin nano-medicines (IDNPs) based on the Tie checkpoint-immune activator prodrug, using the hydrophobicity of IDOi-DON to induce the self-assembly of human serum albumin (HSA) molecules to prepare functional Protein nanomedicine.

The preparation method of the protein nano-medicine utilizes the hydrophobicity of the obtained IDOi-DON molecules, and can induce self-assembly of albumin molecules under the action of an external force to prepare IDNPs functional protein nano-medicines. Include the following steps:

(1) Dissolve the chemically synthesized IDOi-DON molecule in a small amount of organic solvent, and slowly drop the IDOi-DON solution into the aqueous solution of HSA under the continuous action of external force, such as ultrasound, stirring, and vortex oscillation, to induce albumin Molecular self-assembly. The organic solvents include volatile organic solvents and non-volatile organic solvents.

(2) Purify the nanoparticle solution obtained in the first step, remove free drug molecules, and obtain the functional protein nanoparticle solution. After filtering through a 220nm filter membrane, the IDNPs functional protein nanomedicine solution is obtained. The purification means include dialysis, ultrafiltration and centrifugation.

The present invention also provides an in vitro IDOi-DON albumin nano-medicine (IDNPs) anti-tumor verification experiment. In the in vitro CD19 ⁺ SPCA1 tumor model, tumor cells were co-incubated with IDNPs nanoparticles for 72 hours, and glutathione in tumor cells Under the action of peptides, IDNPs were decomposed into IDOi and DON, which synergistically killed tumor cells.

The indoleamine-2,3-dioxygenase inhibitor in the following examples is 1-methyltryptophan (1-MT) as an example, and the metabolic reprogramming agent is 6-diazo-5-oxo- L-norleucine (DON) was taken as an example to demonstrate the preparation method of human serum albumin nanomedicines (IDNPs) based on metabolic checkpoints of the present invention, due to the indoleamine-2,3-dioxygenase inhibitor There are similar structures between metabolic reprogramming agents, and those skilled in the art can deduce and use other indoleamine-2,3-dioxygenase inhibitors according to the method in the embodiment of the present invention Agents and metabolic reprogramming agents can also achieve the purpose of preparing the nanomedicine.

Example 1 Synthesis of IDOi-DON protected by 9-fluorenylmethoxycarbonyl

The specific preparation method is as follows:

Weigh 880 mg of 9-fluorenylmethoxycarbonyl-1-methyltryptophan (2.06 mmol, 1.1 equiv) and 854 mg of HBTU (2.25 mmol, 1.2 equiv) in 14 mL of dry N,N-dimethyl formamide (DMF), then sequentially added 980 μL of diisopropylethylamine (727 mg, 5.63 mmol, 3 equivalents) and isopropyl-DON (400 mg, 1.88 mmol, 1 equivalent) in dry DMF solution, under inert gas atmosphere Stirring was continued for 4 hours.

DMF was removed by rotary evaporation, 100 mL of dichloromethane (DCM) was added, and the resulting organic solution was washed with 100 mL of saturated sodium bicarbonate (NaHCO ₃ ) solution, water, 1 mol/L hydrochloric acid (HCl), water and saturated sodium chloride (NaCl) The solution was washed and dried over anhydrous magnesium sulfate. Evaporation of DCM column chromatography (separation solvent: dichloromethane/ethyl acetate = 5/1) separation and purification to obtain a light yellow solid product, which is 9-fluorenylmethoxycarbonyl-IDOi-DON, the synthetic route is shown in the figure below Show.

Example 2 Preparation of IDOi-DON by removing the carbonyl group of 9-fluorenylmethyloxy group

Weigh 9-fluorenylmethoxycarbonyl-IDOi-DON (900 mg, 2.07 mmol, 1 eq) and dissolve it in 10 mL of dry DCM, slowly add piperidine (514 μL, 5.17 mmol, 2.5 eq) dropwise, and then continue at Stir at room temperature for 4 hours.

The organic solvent was removed by rotary evaporation and vacuum pump, and purified by column chromatography (separation solvent: chloroform/methanol=30/1) to obtain the yellow product IDOi-DON. The synthetic route is shown in the figure below.

Characterize the obtained IDOi-DON, and its H NMR spectrum is shown in Figure 1, in which the peak at ~3.7ppm belongs to -CH ₃ on the indole ring, and the peak at ~1.3ppm belongs to the two isopropyl groups. _-CH3 . The mass spectrum of IDOi-DON is shown in Figure 2, wherein 414m/z belongs to the protonation peak of the prepared IDOi-DON.

The preparation of the human serum albumin nano drug (IDNPs) of embodiment 3 IDOi-DON

Specifically include the following steps:

(1) Dissolve the chemically synthesized IDOi-DON molecule in a small amount of organic solvent, and ultrasonicate the aqueous solution of human serum albumin (HSA) for 5 min using a VCX130 ultrasonic breaker, during which time the IDOi-DON solution is added dropwise to induce albumin molecules to self-sustain. Assemble.

(2) Transfer the solution obtained in step (1) to a 10kDa ultrafiltration tube, centrifuge, wash with deionized water several times to remove free drug molecules, and obtain a functional protein nanoparticle solution that is filtered through a 220nm filter membrane to obtain IDNPs functional protein nanoparticles. Drug solution.

In this example, the nanoparticles were characterized and identified. As shown in Figure 3, the transmission electron microscope image of IDNPs shows a complete spherical structure, and the surface particle size of the dynamic light scattering test result is about 100nm, which is consistent with the electron microscope result.

Example 4 IDNPs killing experiment on tumor cells

1 × 10 ⁶ cultured SPCA1 cells were seeded into a 96-well culture plate, incubated at 37°C and 5% CO ₂ for 24 h, and IDNPs solutions of different concentrations and 1 mM and 2 mM glutathione (GSH) were added in groups. ), and continued co-incubation for 72 h, the cell proliferation was detected by CCK-8 kit. As shown in Figure 4, under the action of GSH, IDNPs nanomedicine exhibited tumor cell growth inhibition, and the inhibition efficiency was proportional to the concentration of IDOi-DON in IDNPs.

The invention discloses a human serum albumin nano drug (IDNPs) based on a metabolic checkpoint, a preparation method and an application thereof. The prodrug molecule IDOi-DON obtained by coupling the indoleamine-2,3-dioxygenase inhibitor (IDOi) and the metabolic reprogramming agent DON through chemical bond amidation reaction can be enzymatically broken in the tumor microenvironment. Coating natural biocompatible serum albumin can improve the stability of the drug, reduce the side effects of the drug, and enrich in the tumor site. Through the synergistic effect of the two drugs, it can inhibit the IDO activity of tumor cells and inhibit the proliferation of Treg cells Simultaneously activate the effect of CD8 ⁺ T cells, synergistically enhance the effect of anti-tumor immunotherapy.

The above descriptions are only preferred embodiments of the present invention, and are not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention shall be included in the protection of the present invention. within range.

Claims

A human serum albumin nanomedicine based on a metabolic checkpoint, characterized in that it includes a prodrug molecular core and human serum albumin coated on the outside, and the prodrug molecule is indoleamine-2,3-dioxygenated A coupling molecule of an enzyme inhibitor and a metabolic reprogramming agent, the coupling mode is amide bond coupling.
The human serum albumin nanomedicine based on metabolic checkpoints according to claim 1, wherein the metabolic reprogramming agent is a glutamine antagonist.
The human serum albumin nanomedicine based on metabolic checkpoints according to claim 2, wherein the metabolic reprogramming agent is selected from the group consisting of 6-diazo-5-oxo-norleucine, 5-diazo -4-oxo-L-norvaline, L-(α-S,5S)-α-amino-3-chloro-4,5-dihydro-5-isoxazoleacetic acid, azaserine any kind.
The human serum albumin nanomedicine based on a metabolic checkpoint according to claim 1, wherein the indoleamine-2,3-dioxygenase inhibitor is an immune activator.
The human serum albumin nanomedicine based on a metabolic checkpoint according to claim 4, wherein the immune activator is 1-methyl tryptophan.
The human serum albumin nanomedicine based on metabolic checkpoints according to claim 1, wherein the human serum albumin coats the prodrug molecular core through hydrophobic interaction.
The human serum albumin nanomedicine based on metabolic checkpoint according to claim 1, wherein the structure of the prodrug molecular core is as shown in formula (I):

Wherein, n is any one of 0, 1, 2, 3, 4, 5, 6, 7; R 1 and R 2 are selected from H or alkyl, substituted alkyl, cycloalkyl, substituted cycloalkane radical, aryl, substituted aryl, heterocyclyl, substituted heterocyclyl, alkenyl, substituted alkenyl, cycloalkenyl, substituted cycloalkenyl, heteroaryl, substituted heteroaryl; R is H or is capable of forming an amide bond, a carbamate bond, a phosphoramidate bond, a phosphoramidate bond with the nitrogen adjacent to R3 ; R3 ' is selected from H or C1-C8 alkyl, substituted C1- C8 alkyl, or R 3 and R 3 ' together form a ring structure comprising -C(=O)-GC(=O)-, wherein G is selected from the group consisting of: C1-C8 alkylene, C1-C8 Heteroalkylene, C5-C8 cycloalkylene, C6-C12 arylene, C5-C12 heteroarylene, divalent C4-C10 heterocycle, each of which may be optionally substituted.
The preparation method of human serum albumin nano-medicine based on metabolic checkpoints according to claim 1, characterized in that it comprises the steps of:

(1) Synthesis of 9-fluorenylmethoxycarbonyl-protected (indoleamine-2,3-dioxygenase inhibitor)-metabolic reprogramming agent;

(2) Preparation of (indoleamine-2,3-dioxygenase inhibitor)-metabolic reprogramming agent prodrug molecule by removing the carbonyl group of 9-fluorenylmethoxy;

(3) Preparation of (indoleamine-2,3-dioxygenase inhibitor)-metabolic reprogramming agent human serum albumin nanomedicine.
The preparation method according to claim 8, characterized in that, in the step (1), isopropyl-metabolic reprogramming agent and 9-fluorenylmethoxycarbonyl-indoleamine-2,3-dioxygenate Coupling of enzyme inhibitors to 9-fluorenylmethoxycarbonyl in the presence of benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate and diisopropylethylamine Protected indoleamine-2,3-dioxygenase inhibitors-metabolic reprogramming agents.
The preparation method according to claim 8, characterized in that, in the step (2), the product obtained in the step (1) is reacted overnight under the action of piperidine to remove the carbonyl group of 9-fluorenylmethoxy to obtain the the prodrug molecules.
The preparation method according to claim 8, characterized in that, in the step (3), (1) dissolves the prodrug molecule obtained in the step (2) in an organic solvent, and slowly dissolves the The prodrug molecule solution is dropped into the aqueous solution of human serum albumin to induce the self-assembly of human serum albumin molecules, and then purify and filter to obtain the human serum albumin nano-medicine based on the metabolic checkpoint.
Application of the human serum albumin nano-medicine based on metabolic checkpoint according to any one of claims 1-7 in tumor immunotherapy.