CN107903267B - Azo aromatic nitrogen mustard-chloroethyl nitrosourea coupled compound, preparation method and application - Google Patents
Azo aromatic nitrogen mustard-chloroethyl nitrosourea coupled compound, preparation method and application Download PDFInfo
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- C07D473/00—Heterocyclic compounds containing purine ring systems
- C07D473/02—Heterocyclic compounds containing purine ring systems with oxygen, sulphur, or nitrogen atoms directly attached in positions 2 and 6
- C07D473/18—Heterocyclic compounds containing purine ring systems with oxygen, sulphur, or nitrogen atoms directly attached in positions 2 and 6 one oxygen and one nitrogen atom, e.g. guanine
Abstract
The present invention relates to compounds or pharmaceutically acceptable salts of the structure of formula (I):
in the compound, the azo group is taken as a hypoxia activation pharmacophore, and the nitrogen-nitrogen double bond of the azo group is broken to release aromatic nitrogen mustard and O under the condition of tumor hypoxia6-BG analogues, which targetedly act as alkylating agents and AGT inhibitors in hypoxic regions, rendering tumor cells more sensitive to alkylating agents; furthermore, the CENUs pharmacophore in the compound can be decomposed to generate chloroethyl carbonium ions, so that the DNA interstrand cross-linking is caused, and the effect of inhibiting the growth of tumor cells is achieved.
Description
Technical Field
The invention relates to the field of pharmacy, in particular to a synthesis method of an aromatic nitrogen mustard-chloroethyl nitrosourea coupling molecule targeting tumor hypoxia and application thereof in preparing an anti-tumor medicament.
Background
The double β -chloroethyl amino group in the nitrogen mustard drug is a functional group which plays a role in resisting tumors, and the group is connected with different carrier structures to form aliphatic nitrogen mustard, aromatic nitrogen mustard, amino acid nitrogen mustard, steroid nitrogen mustard, heterocyclic nitrogen mustard and the like.
Chloroethylnitrosoureas (CENUs) are commonly used clinically for the treatment of brain tumors, myelomas, malignant melanomas, malignant lymphomas and leukemias. CENUs can decompose under physiological conditions to produce chloroethyl carbonium ions, which first react with DNA guanine O6Alkylation of the positions and further reaction with complementary cytosine finally form DNA interstrand cross-links between the N1 position of guanine and the N3 position of cytosine. This DNA interstrand cross-linking hinders the process of DNA replication and transcription in tumor cells, ultimately inducing apoptosis in tumor cells. However, O6the-alkylguanine-DNA Alkyltransferase (AGT) -mediated resistance of CENUs severely hampers the further development of this class of drugs. AGT is a DNA repair protein ubiquitous in cells and capable of converting DNA guanine O6The alkyl groups such as methyl, chloroethyl, benzyl and benzyl at the position are removed, thereby repairing the alkylation damage of the DNA. Studies have shown that AGT results in O via the repair of CENUs6Chloroethylguanine and N1, O6-ethanoguanine, thereby blocking the further reaction of these two alkylated products with cytosine to form DNA inter-strand cross-links, eventually leading to CENUs being unable to effectively inhibit tumor cells and developing resistance.
In order to block AGT-mediated drug resistance and improve the chemotherapeutic effect of CENUs, the design and development of AGT inhibitors with high activity have attracted extensive attention. O is6-benzylguanine (O)6-BG) and Lomegutatrib (Patrin-2) both effective in inhibiting AGT activity and bothThe pseudo substrate of AGT acts with AGT in tumor cells, and blocks O caused by the AGT on CENUs6The repair of alkylguanines, thereby increasing the sensitivity of the tumor cells to CENUs and finally eliminating or reducing the resistance to CENUs. However, the existing AGT inhibitor has no tumor targeting, so that the AGT inhibitor inhibits the activity of AGT in tumor cells and also inhibits the activity of AGT in normal cells, and the toxic and side effects of CENUs are obviously enhanced. Therefore, the design of novel tumor-targeted CENUs or AGT inhibitors is the key to solving the above problems.
The hypoxic microenvironment is an important characteristic that most solid tumors are distinguished from normal tissues, and a new therapeutic strategy for exploring tumor hypoxia has become one of the hot spots of tumor targeted therapy. The Hypoxia Activated Prodrugs (HAPs) are a novel compound designed and synthesized based on tumor hypoxia, are nontoxic or low in toxicity to normal tissues, and can release a pharmacophore with cytotoxicity in a tumor hypoxia area, so that targeted therapy of tumors is realized. Hypoxia-activated prodrugs mainly include nitro-aromatic heterocyclic compounds, N-oxides, quinones, transition metal complexes and compounds containing azo groups. The invention designs and synthesizes a novel combined CENUs prodrug simultaneously containing an AGT inhibitor pharmacophore, an aromatic nitrogen mustard pharmacophore and a hypoxia activation group by utilizing the characteristic that tumor tissues have a hypoxia microenvironment.
Disclosure of Invention
The invention aims to provide an azo aromatic nitrogen mustard-chloroethyl nitrosourea coupled compound or a pharmaceutically acceptable salt. The compound takes azo group as hypoxia activation pharmacophore, and the nitrogen-nitrogen double bond of the azo group is broken to release aromatic nitrogen mustard and O under the condition of tumor hypoxia6-BG analogues, which targetedly act as alkylating agents and AGT inhibitors in hypoxic regions, rendering tumor cells more sensitive to alkylating agents; furthermore, the CENUs pharmacophore in the compound can be decomposed to generate chloroethyl carbonium ions, so that the DNA interstrand cross-linking is caused, and the effect of inhibiting the growth of tumor cells is achieved.
The compound or the pharmaceutically acceptable salt of the invention has a structural formula shown as (I):
r is H, NH2，(CH2)nNH2，CH3，CH2CH3，CH2CH2CH3，CH2OH， CH2CH2OH，NHCOCH3，(CH2)nNHCOCH3，N(CH3)2，(CH2)nN(CH3)2One of (1);
n is an integer of 2 to 6.
Preferably, R is H, NH2，N(CH3)2，NHCOCH3When the compound or the salt thereof is used, the antitumor activity and selectivity of the compound or the salt thereof are high.
Most preferably, when R is H and n is 2 or 3, respectively, i.e., has the following structural formula, the antitumor activity and selectivity of the compound or a salt thereof are most preferable.
Any of the above compounds can be salified with an organic or inorganic acid by means of a technique which is conventional in the art, wherein the salt is preferably hydrochloride, hydrobromide, hydroiodide, sulfate, bisulfate, phosphate, acetate, propionate, butyrate, lactate, methanesulfonate, p-toluenesulfonate, maleate, benzoate, succinate, tartrate, citrate, fumarate, taurate or gluconate, and the salified compound has the same pharmacological activity as the non-salified compound.
The second purpose of the invention is to provide a pharmaceutical composition, which comprises a therapeutically effective amount of the compound with the structure of the formula (I) or pharmaceutically acceptable salt, and at least one pharmaceutical carrier.
The medicinal carrier is a substance commonly used in the field, such as a disintegrating agent, a dispersing agent, a lubricating agent, an emulsifying agent, a stabilizing agent and the like. The pharmaceutical composition can be prepared into pharmaceutically common dosage forms such as tablets, injections, capsules and the like by adopting the conventional means in the field.
A third object of the present invention is to provide a process for the preparation of the compound or pharmaceutically acceptable salt of the present invention, wherein the reaction scheme of the process at least comprises:
(1) reacting the compound a with dihalogenated alkane to obtain a compound b;
(2) reacting the compound b with phthalimide to obtain a compound c;
(3) sequentially adding NaNO into the compound c2Reacting sulfamic acid and N, N-dihydroxyethylaniline to obtain a compound d;
(4) reacting the compound d with thionyl chloride to obtain a compound e;
(5) reacting the compound e with hydrazine hydrate to obtain a compound f;
(6) reacting the compound f with 2-chloroethyl isocyanate to obtain a compound g;
(7) and reacting the compound g with nitrosonium tetrafluoroborate to obtain a compound h.
Preferably, the method comprises the following steps:
(1) compound a with dihaloalkanes according to 1: (1-7) reacting at 40-60 ℃ under the catalysis of alkali to obtain a compound b;
(2) compound b with phthalimide according to 1: (1-4) reacting at 35-70 ℃ to obtain a compound c;
(3) according to the compound c, NaNO2The mol ratio of the sulfamic acid to the N, N-dihydroxyethylaniline is 1: (1-4): (1-5): (3-10) adding NaNO to the Compound c2In N at2Reacting at 0-10 deg.C for 0-1h under protection, adding sulfamic acid, and reacting at 0-10 deg.CReacting for 0-1h, finally adding N, N-dihydroxyethylaniline, and reacting for 1-4h at the temperature of 0-10 ℃ to obtain a compound d;
(4) compound d with SOCl2According to the following steps of 1: (1-7) carrying out reflux reaction at the temperature of 20-40 ℃ to obtain a compound e;
(5) reacting the compound e with hydrazine hydrate at the temperature of 25-60 ℃ to obtain a compound f;
(6) compound f with 2-chloroethyl isocyanate according to 1: (1-8) reacting at 0-10 ℃ to obtain a compound g;
(7) compound g was reacted with nitrosonium tetrafluoroborate according to 1: (1-5) reacting at 0-10 ℃ to obtain a compound h.
Preferably, in step (1), the dihaloalkane is a dibromoalkane or a dichloroalkane; dibromoalkanes are preferred. The dibromoalkane has higher reaction activity, can ensure the smooth reaction, and obtains higher yield and purer products.
Further preferred is:
the specific operation of the step (1): dissolving the compound a, adding catalyst inorganic base into the solution, dropwise adding 1, 2-dibromo alkane into the solution, and reacting at the temperature of 40-60 ℃ to obtain a compound b;
wherein, the mol ratio of the compound a, the inorganic base and the 1, 2-dibromo alkane is preferably 1: (1-6): (1-7); the inorganic base is preferably anhydrous potassium carbonate; the solvent for dissolving the compound a is preferably acetone or N, N-dimethylformamide; the step (1) further comprises a step of purifying the compound b, specifically concentrating the reaction solution, and separating and purifying the concentrated solution by adopting a column chromatography, wherein preferably a stationary phase of the column chromatography is silica gel, a mobile phase of the column chromatography is petroleum ether and ethyl acetate, and further preferably the compound b is purified by gradient elution according to a ratio that the petroleum ether/ethyl acetate (v/v) ratio is gradually changed from 1:2 to 1: 4.
The specific operation of the step (2) is as follows: compound b with phthalimide according to 1: (1-4) reacting at 40-70 ℃ to obtain a compound c. Wherein, the reaction solvent used in the step (2) is preferably acetonitrile or N, N-dimethylformamide; the step (2) further comprises a step of purifying the compound c, and specifically comprises the steps of adding ethyl acetate and water (v: v ═ 1:1) into the reaction solution for extraction, collecting an organic phase, washing with a saturated aqueous sodium chloride solution, drying, and distilling to remove the solvent.
The specific operation of the step (3) is as follows: according to the compound c, NaNO2The mol ratio of the sulfamic acid to the N, N-dihydroxyethylaniline is 1: (1-4): (1-5): (3-10), NaNO was added to the Compound c2，N2Under protection, reacting for 10-30 min at 0-10 ℃, then adding sulfamic acid for reacting for 10-20 min at 0-10 ℃, and then reacting with N, N-dihydroxyethylaniline for 1.5-2.5 h at 0-10 ℃ to obtain a compound d
The specific operation of step (3) is preferably: dissolving the compound c, and dropwise adding and dissolving NaNO at 0 ℃ under the protection of inert gas2Reacting for 10-30 min, then dropwise adding a solution for dissolving sulfamic acid, reacting for 10-20 min, then dropwise adding a solution containing N-N dihydroxyethylaniline into the reaction solution, and carrying out ice bath at 0 ℃ for 3h to obtain a compound d.
Wherein, the solvent for dissolving the compound c is preferably a mixed solution of acetonitrile, dichloromethane, trifluoroacetic acid and water; and (3) further comprising a step of purifying the compound d, specifically concentrating the reaction solution, and separating and purifying the concentrated solution by adopting a column chromatography, wherein preferably a stationary phase of the column chromatography is silica gel, a mobile phase of the column chromatography is acetone and dichloromethane, and further preferably gradient elution is carried out according to a ratio that acetone/dichloromethane (v/v) is gradually changed to 1:2 to purify the compound e.
The specific operation of the step (4) is as follows: according to the compounds d, SOCl2In a molar ratio of 1: (1-7) refluxing for 1.5-2.5 h at 20-40 ℃ to obtain a compound e. Among them, the solvent for dissolving the compound d is preferably dichloromethane. And (4) purifying the compound e, specifically concentrating the reaction solution, and separating and purifying the concentrated solution by adopting a column chromatography, wherein preferably a stationary phase of the column chromatography is silica gel, a mobile phase of the column chromatography is methanol and dichloromethane, and further preferably the compound e is purified by gradient elution according to a ratio that methanol/dichloromethane (v/v) is gradually changed from 1:2 to 1: 4.
The specific operation of the step (5) is as follows: reacting the compound e with hydrazine hydrate at 25-60 ℃ to obtain a compound f, wherein the molar volume ratio of the compound e to 70% of hydrazine hydrate is (1-5): 1 (mmol: mL). Firstly, dissolving a compound e, and then reacting the compound e with hydrazine hydrate, wherein a solvent for dissolving the compound e is preferably DMF or methanol; the step (4) further includes a step of purifying the compound f, specifically, the compound f is obtained by adding dichloromethane and water (v: v ═ 1:1) to the reaction solution to perform extraction, collecting an organic phase, washing with a saturated aqueous sodium chloride solution, drying, and distilling off the solvent.
The specific operation of the step (6) is as follows: compound f with 2-chloroethyl isocyanate according to 1: (1-8) at 0-10 ℃ to obtain a compound g. Among them, dichloromethane is preferable as the reaction solvent, and after completion of the reaction, the solvent is distilled off to obtain a compound g.
The specific operation of the step (7) is as follows: compound g, glacial acetic acid, nitrosonium tetrafluoroborate according to 1: (1-5): and (1-5) reacting under ice bath condition to obtain a compound h.
The compound h in the invention is a compound with a structure shown in a formula (I).
Among them, the reaction solvent of step (7) is preferably acetonitrile or acetone. And (6) further comprising a step of purifying the compound h, specifically, after the reaction is finished, adding ethyl acetate and water (v: v ═ 1:1) into the reaction solution for extraction, washing an organic phase, drying and concentrating, and separating the concentrated solution by adopting column chromatography, wherein the column chromatography preferably adopts silica gel as a stationary phase and petroleum ether and ethyl acetate as mobile phases, and further preferably adopts a gradient that the petroleum ether/ethyl acetate (v/v) gradient is gradually changed from 1:1 to 1:5 to obtain the compound h.
The fourth purpose of the invention is to protect the application of the compound, the pharmaceutically acceptable salt or the composition in preparing the antitumor drugs.
Preferably, the tumor is one or more of brain tumor, myeloma, malignant melanoma, malignant lymphoma, lung cancer, breast cancer, gastric cancer, colon cancer, prostate tumor, leukemia and lymph cancer;
more preferably one or more of brain tumor, leukemia, lung cancer, colon cancer, and lymph cancer cell.
The compound or pharmaceutically acceptable salt in the general formula is a novel aromatic nitrogen mustard combined chloroethyl nitrosourea with drug resistance and tumor cell targeting, and the compound in the general formula (I) is subjected to an in-vitro anti-tumor screening test. The results show that the compound in the general formula (I) has obvious inhibition effects on various tumor cell lines such as L1210 mouse leukemia cells, HT29 human colon cancer cells, A549 human lung cells, HUT102 human T lymphoma cells, human glioma cells SF763, SF126 and the like under the hypoxia condition, while the inhibition effects on the tumor cells by the compound in the general formula (I) are not obvious under the aerobic condition. Therefore, the compound in the general formula (I) has good targeting property and capability of killing tumor cells, and can be used for targeted tumor chemotherapy drugs. The compound can release aromatic nitrogen mustard and O by breaking nitrogen-nitrogen double bond of azo group under tumor hypoxia condition6-BG analogues, which targetedly act as alkylating agents and AGT inhibitors in hypoxic regions, rendering tumor cells more sensitive to alkylating agents; the compound is also a derivative of chloroethylnitrosourea, and is decomposed to generate chloroethyl carbonium ions, so that cross-linking among DNA strands is generated, and the effect of inhibiting the growth of tumor cells is exerted.
The compounds of the present invention have three advantages: firstly, the azobenzene structure enables the compound to have low oxygen selectivity; second, the hypoxic region releases O6-BG analogues confer resistance to the compound targeting tumor hypoxia; thirdly, the combined action of CENUs and aromatic nitrogen mustard double alkylating agent functional groups makes the antitumor activity of the compound higher.
Detailed Description
The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention. The medicines involved in the following examples are not specifically described, and are commercially available; the operations involved, unless otherwise specified, are those conventional in the art.
The structural formulae of compound 1 and compound 2 referred to in the following examples are as follows:
the following examples are intended to illustrate the invention but are not intended to limit the scope of the invention.
Example 1: (E) synthesis of (E) -3- (2- (6- (benzyloxy) -2- ((4- (bis (2-chloroethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) ethyl) -1- (2-chloroethyl) -1-nitrosourea (Compound 1) N9-bromoethyl-O6Synthesis of-benzylguanine
Weighing O6Adding benzyl guanine (2.05g,8.5mmol) and anhydrous potassium carbonate (3.35g, 24.3mmol) into a 100mL round-bottom flask, adding 50mL acetone, slowly heating to 52 ℃, dropwise adding 1, 2-dibromoethane (2.89mL,33mmol), continuing to react for 72h after dropwise adding, filtering the reaction solution, collecting the filtrate, distilling at 40 ℃ under reduced pressure to remove the solvent, separating and purifying by silica gel column chromatography, using petroleum ether and ethyl acetate as eluent, gradually increasing the volume ratio of the petroleum ether to the ethyl acetate from 1:2 to 1:4 by gradient elution, and drying at 40 ℃ in vacuum to obtain a white solid N9-bromoethyl-O6Benzylguanine (2.05g,5.9mmol), yield 69%.
UVλ:48,283nm；
IR (KBr pellet) v/cm-1:3467.1(-(CH2)2-N-H)；3318.8(N-H)；2957.3 (-CH2-)；1633.6(C＝C)；1249.9(C-O-C)；1065.7(C-N)；687.8(C-Br)；
1H NMR(400MHz,CDCl3)δ:3.77(t,2H,CH2)；4.21(t,2H,CH2)； 5.18(s,2H,CH2)；6.95(s,2H,NH2)；7.35-7.49(m,5H,C6H5)；8.13(s,1H, CH)；
ESI-MS:m/z348(M+H)+。
2) N9- (2- (N-phthalimido) ethyl) -O6-benzylguanine
The solid obtained was N9-bromoethyl-O6Benzyl guanine (2.05g,5.9mmol), 15mL anhydrousDissolving N, N-dimethylformamide, adding potassium phthalimide solid (3.42g,18.5mmol), heating to 65 deg.C, stirring for reaction for 6h, extracting with ethyl acetate and deionized water, washing the organic phase with saturated sodium chloride aqueous solution, drying with anhydrous sodium sulfate overnight, and vacuum drying at 35 deg.C to obtain white solid N9- (2- (N-phthalimido) ethyl) -O6Benzylguanine (2.07g,5mmol), yield 85%.
UVλ:228,284nm；
IR (KBr pellet) v/cm-1:3492.4(-(CH2)2-N-H；2955.9(-CH2-)；1723.9 (C＝O)；1621.9(C＝C)；1256.9(C-O-C)；1069.1(C-N)；
1H NMR(400MHz,CDCl3)δ:4.14(t,2H,CH2)；4.65(t,2H,CH2)； 5.12(s,2H,CH2)；6.95(s,2H,NH2)；7.35-7.46(m,5H,C6H5)；7.81-7.89 (m,4H,C6H4)；8.06(s,1H,CH)；
ESI-MS:m/z415(M+H)+。
3) Synthesis of (E) -2- (2- (6- (benzyloxy) -2- ((4- (bis (2-hydroxyethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) ethyl) isoindoline-1, 3-dione
First, 25mL of a mixture (5mL of acetonitrile, 20mL of dichloromethane, 50uL of trifluoroacetic acid, and 25uL of water) was prepared. At 0 ℃ N2Under protection of N9- (2- (N-phthalimido) ethyl) -O in a downward stirring manner6To a 21mL mixture of (2.07g,5mmol) of (E) -benzylguanine, NaNO was added2(0.69g, 10mmol), and reacted for 20 min. Then, 4mL of a mixture containing sulfamic acid (0.92g, 9.5mmol) was injected and reacted for 15 min. A further 10mL of acetonitrile containing N-phenyldiethanolamine (6.08g, 36mmol) was injected and reacted at 0 ℃ for 2 h. Extracting with dichloromethane and water, mixing organic layers, and extracting with anhydrous Na2SO4Drying overnight, distilling at 30 deg.C under reduced pressure to remove solvent, separating and purifying with silica gel column chromatography, eluting with acetone and dichloromethane, gradually changing the volume ratio of acetone/dichloromethane from 1:19 to 1:2 by gradient elution, and vacuum drying at 30 deg.C to obtain white solid (E) -2- (2- (6- (benzyloxy) -2- ((4- (bis (2-hydroxyethyl) amino) phenyl) diazene)Yl) -9H-purin-9-yl) ethyl) isoindoline-1, 3-dione (1.94g,3.2mmol), 64% yield.
UVλ:224,275nm；
IR (KBr pellet) v/cm-1:3630.1(-OH)；3489.4(N-(CH2)2-)；3327.9 (N-H)；3174.9(C-H)；1797.4(C＝O)；1753.4(N＝N)；1645.1(C＝C)；265.5 (C-O-C)；1086.7(C-N)；
1H NMR(400MHz,CDCl3)δ:3.59(s,1H,OH)；3.69(s,1H,OH)； 3.72(t,2H,CH2)；3.73(t,2H,CH2)；4.15(t,2H,CH2)；4.22(t,2H,CH2)；4.23 (t,2H,CH2)；4.63(t,2H,CH2)；5.19(s,2H,CH2)；6.72-7.76(m,4H,C6H4)； 7.32-7.48(m,5H,C6H5)；7.85-7.89(m,4H,C6H4)；8.11(s,1H,CH)；
ESI-MS:m/z607(M+H)+。
4) Synthesis of (E) -2- (2- (6- (benzyloxy) -2- ((4- (bis (2-chloroethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) ethyl) isoindoline-1, 3-dione
To a solution of 1.94g (3.2mmol) of (E) -2- (2- (6- (benzyloxy) -2- ((4- (bis (2-hydroxyethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) ethyl) isoindoline-1, 3-dione in 20mL dichloromethane was added 1.4mL (20mmol) SOCl2The reaction was carried out at 30 ℃ for 1.5 hours under reflux. Extracting with dichloromethane and water, mixing organic layers, and extracting with anhydrous Na2SO4Drying overnight at 30 deg.C under reduced pressure to remove solvent, separating and purifying by silica gel column chromatography using methanol and dichloromethane as eluent, gradually changing the volume ratio of methanol/dichloromethane from 1:2 to 1:4 by gradient elution, and vacuum drying at 30 deg.C to obtain white solid (E) -2- (2- (6- (benzyloxy) -2- ((4- (bis (2-chloroethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) ethyl) isoindoline-1, 3-dione (1.35g,2.1mmol) with 66% yield.
UVλ:221,272nm；
IR (KBr pellet) v/cm-1:3489.4(N-(CH2)2-)；3327.9(N-H)；3174.9 (C-H)；1797.4(C＝O)；1748.4(N＝N)；1645.1(C＝C)；1382.4(N-O)； 1265.5(C-O-C)；1086.7(C-N)；775.4(C-Cl)；
1H NMR(400MHz,CDCl3)δ:3.58(t,2H,CH2)；3.61(t,2H,CH2)； 3.65(t,2H,CH2)；3.66(t,2H,CH2)；4.11(t,2H,CH2)；4.63(t,2H,CH2)；5.16 (s,2H,CH2)；6.78-7.74(m,4H,C6H4)；7.39-7.47(m,5H,C6H5)；7.85-7.88 (m,4H,C6H4)；8.05(s,1H,CH)；
ESI-MS:m/z643(M+H)+。
5) Synthesis of (E) -4- ((9- (2-aminoethyl) -6- (benzyloxy) -9H-purin-2-yl) diazenyl) -N, N-bis (2-chloroethyl) aniline
The resulting solid (E) -2- (2- (6- (benzyloxy) -2- ((4- (bis (2-chloroethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) ethyl) isoindoline-1, 3-dione (1.35g,2.1mmol) was dissolved in 10mL of DMF (N, N-dimethylformamide) and 2.4mL of hydrazine hydrate was added, the reaction was stirred at 32 ℃ for 4H to hydrazinolysis, extracted with dichloromethane and deionized water, the organic phase was washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate overnight, dried at 30 ℃ in vacuo to give (E) -4- ((9- (2-aminoethyl) -6- (benzyloxy) -9H-purin-2-yl) diazenyl) -N, n-bis (2-chloroethyl) aniline (0.82g,1.6mmol), yield 76%.
UVλ:226,276nm；
IR (KBr pellet) v/cm-1:3457.3(N-(CH2)2-)；3327.9(N-H)；3174.9 (C-H)；1789.5(C＝O)；1747.4(N＝N)；1692.6(C＝C)；1426.4(N-O)； 1288.7(C-O-C)；1097.4(C-N)；772.3(C-Cl)；
1H NMR(400MHz,CDCl3)δ:3.14(t,2H,CH2)；3.61(t,2H,CH2)； 3.63(t,2H,CH2)；4.58(t,2H,CH2)；5.14(s,2H,NH2)；6.71-7.74 (m,4H,C6H4)；7.35-7.42(m,5H,C6H5)；8.05(s,1H,CH)；
ESI-MS:m/z513(M+H)+。
6) Synthesis of (E) -2- (2- (6- (benzyloxy) -2- ((4- (bis (2-hydroxyethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) ethyl) isoindoline-1, 3-dione
The resulting solid (E) -4- ((9- (2-aminoethyl) -6- (benzyloxy) -9H-purin-2-yl) diazenyl) -N, N-bis (2-chloroethyl) aniline (0.82g,1.6mmol) was dissolved in 10mL of dichloromethane, 5mL of dichloromethane containing 2-chloroethyl isocyanate (0.68mL,8mmol) was added dropwise, the reaction was carried out in an ice bath at 0 ℃ for 6H, and after completion of the reaction, (E) -2- (2- (6- (benzyloxy) -2- ((4- (bis (2-hydroxyethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) ethyl) isoindoline-1, 3-dione (0.56g,0.9mmol) was obtained by distillation under reduced pressure at 30 ℃, the yield was 57%.
UVλ:224,270nm；
IR (KBr pellet) v/cm-1:3345.7(N-H)；3096.2(C-H)；1772.1(C＝O)； 1759.4(N＝N)；1657.9(C＝C)；1365.4(N-O)；1234.5(C-O-C)； 1120.2(C-N)；769.7(C-Cl)；
1H NMR(400MHz,CDCl3)δ:3.45(t,2H,CH2)；3.62(t,2H,CH2)； 3.64(t,2H,CH2)；4.56(t,2H,CH2)；5.16(s,2H,CH2)；6.2(t,1H,NH)； 6.76-7.74(m,4H,C6H4)；7.38-7.47(m,5H,C6H5)；8.06(s,1H,CH)；
ESI-MS:m/z618(M+H)+。
7) Synthesis of (E) -3- (2- (6- (benzyloxy) -2- ((4- (bis (2-chloroethyl) amino) phenyl) diazenyl) -9H-purine-9-chloroethylnitrosourea (Compound 1)
Dissolving the obtained solid (E) -2- (2- (6- (benzyloxy) -2- ((4- (bis (2-hydroxyethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) ethyl) isoindoline-1, 3-dione (0.56g,0.9mmol) in 8mL acetonitrile, adding 3mL acetonitrile containing glacial acetic acid (11.2. mu.L, 0.2mmol), adding nitrosonium tetrafluoroborate (0.48g,4mmol) under ice bath conditions, reacting for 4H under ice bath conditions, extracting with ethyl acetate and deionized water after completion of the reaction, washing the organic phase with saturated aqueous sodium chloride solution, drying with anhydrous sodium sulfate, distilling off the solvent under reduced pressure, separating and purifying with silica gel column chromatography, wherein the eluent is petroleum ether and ethyl acetate, and gradually increasing the volume ratio of petroleum ether to ethyl acetate from 1:1 to 1:4 by gradient elution, vacuum drying at 30 deg.C gave (E) -3- (2- (6- (benzyloxy) -2- ((4- (bis (2-chloroethyl) amino) phenyl) diazenyl) -9H-purine-9-chloroethylnitrosourea (0.37g,0.58mmol) in 63% yield.
UVλ:225,274nm；
IR (KBr pellet) v/cm-1:3365.6(N-H)；3024.5(C-H)；1765.1(C＝O)； 1755.4(N＝N)；1685.8(C＝C)；1574.5(N＝O)；1372.6(N-O)；1293.4 (C-O-C)；1132.1(C-N)；1065.7(N-N)；772.8(C-Cl)；
1H NMR(400MHz,CDCl3)δ:3.45(t,2H,CH2)；3.62(t,2H,CH2)； 3.63(t,2H,CH2)；4.56(t,2H,CH2)；5.16(s,2H,CH2)；6.3(t,1H,NH)； 6.72-7.77(m,4H,C6H4)；7.35-7.49(m,5H,C6H5)；8.05(s,1H,CH)；
ESI-MS:m/z647(M+H)+。
Example 2: (E) synthesis of (E) -3- (2- (6- (benzyloxy) -2- ((4- (bis (2-chloroethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) ethyl) -1- (2-chloroethyl) -1-nitrosourea (Compound 1) N9-bromoethyl-O6Synthesis of-benzylguanine
Weighing O6Adding benzylguanine (2.12g,8.8mmol) and anhydrous potassium carbonate (3.36g, 24.32mmol) into a 100mL round-bottom flask, adding 50mL acetone, slowly heating to 52 ℃, dropwise adding 1, 2-dibromoethane (3.02mL,34.5mmol), continuing to react for 75 hours after finishing dropping, filtering the reaction solution, collecting the filtrate, distilling the spin-dried solvent at 40 ℃ under reduced pressure, separating and purifying by silica gel column chromatography, gradually increasing the volume ratio of petroleum ether to ethyl acetate from 1:2 to 1:4 by gradient elution, and drying in vacuum at 40 ℃ to obtain a white solid N9-bromoethyl-O6Benzylguanine (2.16g,6.22mmol), yield 71%.
UVλ:248,283nm；
IR (KBr pellet) v/cm-1:3468.2(-(CH2)2-N-H)；3325.1(N-H)；2963.1 (-CH2-)；1638.2(C＝C)；1255.2(C-O-C)；1064.9(C-N)；682.1(C-Br)；
1H NMR(400MHz,CDCl3)δ:3.71(t,2H,CH2)；4.22(t,2H,CH2)； 5.25(s,2H,CH2)；6.91(s,2H,NH2)；7.35-7.45(m,5H,C6H5)；8.19 (s,1H,CH)；
ESI-MS:m/z348(M+H)+。
2) N9- (2- (N-phthalimido) ethyl) -O6-benzylguanine
The solid obtained was N9-bromoethyl-O6Benzyl guanine (2.16g,6.22mmol), dissolving in 15mL of anhydrous N, N-dimethylformamide, adding solid potassium phthalimide (3.46g,18.7mmol), heating to 68 ℃, stirring for reaction for 6.5h, extracting with ethyl acetate and deionized water, washing the organic phase with saturated aqueous sodium chloride solution, drying over night with anhydrous sodium sulfate, and vacuum drying at 35 ℃ to obtain white solid N9- (2- (N-phthalimido) ethyl) -O6Benzylguanine (2.24g,5.42mmol), yield 87%.
UVλ:228,284nm；
IR (KBr pellet) v/cm-1:3498.3(-(CH2)2-N-H)；2957.2(-CH2-)；1725.2 (C＝O)；1622.3(C＝C)；1258.4(C-O-C)；1069.1(C-N)；
1H NMR(400MHz,CDCl3)δ:4.15(t,2H,CH2)；4.66(t,2H,CH2)； 5.17(s,2H,CH2)；6.97(s,2H,NH2)；7.37-7.42(m,5H,C6H5)；7.81-7.85 (m,4H,C6H4)；8.12(s,1H,CH)；
ESI-MS:m/z415(M+H)+。
3) Synthesis of (E) -2- (2- (6- (benzyloxy) -2- ((4- (bis (2-hydroxyethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) ethyl) isoindoline-1, 3-dione
First, 25mL of a mixture (5mL of acetonitrile, 20mL of dichloromethane, 50uL of trifluoroacetic acid, and 25uL of water) was prepared. At 0 ℃ N2Under protection of N9- (2- (N-phthalimido) ethyl) -O in a downward stirring manner6To a 21mL mixture of (2.24g,5.42mmol) of benzylguanine was added NaNO2(0.76g, 11mmol), and reacted for 20 min. Reinjection of ammonia4mL of a mixture of sulfamic acid (1.07g, 11mmol) was reacted for 15 min. A further 10mL of acetonitrile containing N-phenyldiethanolamine (6.39g, 38mmol) was injected and reacted at 0 ℃ for 3 h. Extracting with dichloromethane and water, mixing organic layers, and extracting with anhydrous Na2SO4Drying overnight, distilling off the solvent under reduced pressure at 30 ℃ and separating and purifying by silica gel column chromatography eluting with acetone and dichloromethane with gradient elution and a volume ratio of acetone/dichloromethane gradually changing from 1:19 to 1:2, drying under vacuum at 30 ℃ to obtain (E) -2- (2- (6- (benzyloxy) -2- ((4- (bis (2-hydroxyethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) ethyl) isoindoline-1, 3-dione (2.12g,3.5mmol) as a white solid with a yield of 65%.
UVλ:224,275nm；
IR (KBr pellet) v/cm-1:3639.1(-OH)；3489.4(N-(CH2)2-)；3332.4 (N-H)；3171.5(C-H)；1791.6(C＝O)；1756.2(N＝N)；1648.2(C＝C)； 1265.2(C-O-C)；1089.2(C-N)；
1H NMR(400MHz,CDCl3)δ:3.55(s,1H,OH)；3.64(s,1H,OH)； 3.72(t,2H,CH2)；3.76(t,2H,CH2)；4.16(t,2H,CH2)；4.23(t,2H,CH2)；4.29 (t,2H,CH2)；4.61(t,2H,CH2)；5.20(s,2H,CH2)；6.74-7.78(m,4H,C6H4)； 7.32-7.45(m,5H,C6H5)；7.84-7.89(m,4H,C6H4)；8.12(s,1H,CH)；
ESI-MS:m/z607(M+H)+。
4) Synthesis of (E) -2- (2- (6- (benzyloxy) -2- ((4- (bis (2-chloroethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) ethyl) isoindoline-1, 3-dione
To a solution of 2.12g (3.5mmol) of (E) -2- (2- (6- (benzyloxy) -2- ((4- (bis (2-hydroxyethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) ethyl) isoindoline-1, 3-dione in 22mL dichloromethane was added 1.54mL (22mmol) SOCl2And reacted at 33 ℃ for 2 hours under reflux. Extracting with dichloromethane and water, mixing organic layers, and extracting with anhydrous Na2SO4Drying overnight, distilling off solvent at 30 deg.C under reduced pressure, and separating with silica gel column chromatographyThe eluent was methanol and dichloromethane and the volume ratio methanol/dichloromethane was gradually changed from 1:2 to 1:4 by gradient elution and dried under vacuum at 30 ℃ to give (E) -2- (2- (6- (benzyloxy) -2- ((4- (bis (2-chloroethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) ethyl) isoindoline-1, 3-dione (1.54g,2.4mmol) as a white solid in 69% yield.
UVλ:221,272nm；
IR (KBr pellet) v/cm-1:3495.8(N-(CH2)2-)；3336.4(N-H)；3175.4 (C-H)；1789.8(C＝O)；1752.3(N＝N)；1649.2(C＝C)；1385.1(N-O)； 1268.1(C-O-C)；1089.1(C-N)；772.4(C-Cl)；
1H NMR(400MHz,CDCl3)δ:3.55(t,2H,CH2)；3.64(t,2H,CH2)； 3.66(t,2H,CH2)；3.69(t,2H,CH2)；4.11(t,2H,CH2)；4.62(t,2H,CH2)；5.18 (s,2H,CH2)；6.75-7.74(m,4H,C6H4)；7.38-7.41(m,5H,C6H5)；7.81-7.86 (m,4H,C6H4)；8.12(s,1H,CH)；
ESI-MS:m/z643(M+H)+。
5) Synthesis of (E) -4- ((9- (2-aminoethyl) -6- (benzyloxy) -9H-purin-2-yl) diazenyl) -N, N-bis (2-chloroethyl) aniline
The resulting solid (E) -2- (2- (6- (benzyloxy) -2- ((4- (bis (2-chloroethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) ethyl) isoindoline-1, 3-dione (1.52g,2.4mmol) was dissolved in 12mL of DMF (N, N-dimethylformamide) and 2.6mL of hydrazine hydrate was added, the reaction was allowed to hydrazinolyze with stirring at 35 ℃ for 4.5H, extracted with dichloromethane and deionized water, the organic phase was washed with saturated aqueous sodium chloride, dried over anhydrous sodium sulfate overnight, and dried in vacuo at 30 ℃ to give (E) -4- ((9- (2-aminoethyl) -6- (benzyloxy) -9H-purin-2-yl) diazenyl) -N, n-bis (2-chloroethyl) aniline (0.95g,1.85mmol), 78% yield.
UVλ:226,276nm；
IR (KBr pellet) v/cm-1:3462.2(N-(CH2)2-)；3331.2(N-H)；3176.2 (C-H)；1792.4(C＝O)；1749.4(N＝N)；1693.6(C＝C)；1428.1(N-O)； 1293.1(C-O-C)；1098.1(C-N)；772.5(C-Cl)；
1H NMR(400MHz,CDCl3)δ:3.15(t,2H,CH2)；3.62(t,2H,CH2)； 3.68(t,2H,CH2)；4.59(t,2H,CH2)；5.15(s,2H,NH2)；6.72-7.75 (m,4H,C6H4)；7.36-7.42(m,5H,C6H5)；8.08(s,1H,CH)；
ESI-MS:m/z513(M+H)+。
6) Synthesis of (E) -2- (2- (6- (benzyloxy) -2- ((4- (bis (2-hydroxyethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) ethyl) isoindoline-1, 3-dione
The resulting solid (E) -4- ((9- (2-aminoethyl) -6- (benzyloxy) -9H-purin-2-yl) diazenyl) -N, N-bis (2-chloroethyl) aniline (0.95g,1.85mmol) was dissolved in 12mL of dichloromethane, 5mL of dichloromethane containing 2-chloroethyl isocyanate (0.85mL,10mmol) was added dropwise, the reaction was carried out in an ice bath at 0 ℃ for 6.5H, and after completion of the reaction, (E) -2- (2- (6- (benzyloxy) -2- ((4- (bis (2-hydroxyethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) ethyl) isoindoline-1, 3-dione (0.68g,1.1mmol) was obtained by distillation under reduced pressure at 30 ℃, the yield was 59%.
UVλ:224,270nm；
IR (KBr pellet) v/cm-1:3348.7(N-H)；3094.2(C-H)；1775.2(C＝O)； 1759.4(N＝N)；1658.4(C＝C)；1368.3(N-O)；1235.6(C-O-C)；1120.2 (C-N)；770.2(C-Cl)；
1H NMR(400MHz,CDCl3)δ:3.46(t,2H,CH2)；3.65(t,2H,CH2)； 3.69(t,2H,CH2)；4.56(t,2H,CH2)；5.16(s,2H,CH2)；6.23(t,1H,NH)； 6.72-7.72(m,4H,C6H4)；7.35-7.45(m,5H,C6H5)；8.07(s,1H,CH)；
ESI-MS:m/z618(M+H)+。
7) Synthesis of (E) -3- (2- (6- (benzyloxy) -2- ((4- (bis (2-chloroethyl) amino) phenyl) diazenyl) -9H-purine-9-chloroethylnitrosourea (Compound 1)
Dissolving the obtained solid (E) -2- (2- (6- (benzyloxy) -2- ((4- (bis (2-hydroxyethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) ethyl) isoindoline-1, 3-dione (0.68g,1.1mmol) in 10mL acetonitrile, adding 3mL acetonitrile containing glacial acetic acid (22.4. mu.L, 0.4mmol), adding nitrosonium tetrafluoroborate (0.53g,4.5mmol) under ice bath conditions, reacting for 5H under ice bath conditions, extracting with ethyl acetate and deionized water after completion of the reaction, washing the organic phase with saturated aqueous sodium chloride solution, drying with anhydrous sodium sulfate, distilling off the solvent under reduced pressure, separating and purifying with silica gel column chromatography, eluting with petroleum ether and ethyl acetate by gradient elution, wherein the volume ratio of petroleum ether to ethyl acetate is gradually increased from 1:1 to 1:4, vacuum drying at 30 ℃ gave (E) -3- (2- (6- (benzyloxy) -2- ((4- (bis (2-chloroethyl) amino) phenyl) diazenyl) -9H-purine-9-chloroethylnitrosourea (0.48g,0.75mmol) in 67% yield.
UVλ:225,274nm；
IR (KBr pellet) v/cm-1:3368.2(N-H)；3027.2(C-H)；1768.3(C＝O)； 1758.1(N＝N)；1686.1(C＝C)；1574.2(N＝O)；1375.1(N-O)；1294.2 (C-O-C)；1138.1(C-N)；1072.2(N-N)；776.2(C-Cl)；
1H NMR(400MHz,CDCl3)δ:3.44(t,2H,CH2)；3.60(t,2H,CH2)； 3.65(t,2H,CH2)；4.58(t,2H,CH2)；5.14(s,2H,CH2)；6.34(t,1H,NH)； 6.72-7.72(m,4H,C6H4)；7.36-7.50(m,5H,C6H5)；8.12(s,1H,CH)；
ESI-MS:m/z647(M+H)+。
Example 3: (E) synthesis of (E) -3- (3- (6- (benzyloxy) -2- ((4-bis (2-chloroethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) propyl) -1- (2-chloroethyl) -1-nitrosourea (Compound 2)
1) N9-Bromopropyl-O6Synthesis of-benzylguanine
Weighing O6-benzylguanine (1.45g,6mmol) and anhydrous potassium carbonate (2.76g,20mmol) are added into a 50mL round bottom flask, 50mL acetone is added, the temperature is slowly raised to 55 ℃,1, 3 dibromopropane (3.03mL,30mmol) is dropwise added, the reaction is continued for 65 hours after the dropwise addition is finished, the reaction solution is filtered, the filtrate is collected, the solvent is removed by distillation under reduced pressure at 45 ℃, and then silicon is usedSeparating and purifying by gel column chromatography, eluting with petroleum ether and ethyl acetate, gradient eluting, gradually increasing petroleum ether/ethyl acetate volume ratio from 1:2 to 1:4, and vacuum drying at 40 deg.C to obtain white solid N9-bromopropyl-O6Benzylguanine (1.41g,3.9mmol), yield 65%.
UVλ:250,284nm；
IR (KBr pellet) upsilon/cm-1:3452.4(N-(CH2)3-)；3312.6(N-H)；2947.9 (C-H)；1634.7(C＝C)；1258.9(C-O-C)；1052.7(C-N)；733.9(C-Br)；
1H NMR(400MHz,CDCl3)δ:2.25(m,2H,CH2)；3.55(t,2H,CH2)； 4.08(t,2H,CH2)；5.15(s,2H,CH2)；6.92(s,2H,NH2)；7.38-7.45 (m,5H,C6H5)；8.11(s,1H,CH)；
ESI-MS:m/z362(M+H)+。
2) N9- (3- (N-phthalimido) propyl) -O6Synthesis of-benzylguanine
The solid obtained was N9-bromopropyl-O6-benzylguanine (1.41g,3.9mmol), dissolving in 15mL of anhydrous N, N-dimethylformamide, adding solid potassium phthalimide (1.48g,8mmol), heating to 65 ℃, stirring for reaction for 6h, extracting with ethyl acetate and deionized water, washing the organic phase with saturated aqueous sodium chloride solution, drying over night with anhydrous sodium sulfate, and vacuum drying at 40 ℃ to obtain white solid N9- (3- (N-phthalimido) propyl) -O6Benzylguanine (1.24g,2.9mmol), yield 74%.
UVλ:225,286nm；
IR (KBr pellet) v/cm-1:3482.7(N-(CH2)3-)；3341.1(N-H)；2975.4 (C-H)；1783.9(C＝O)；1631.7(C＝C)；1272.6(C-O-C)；1087.7(C-N)；
1H NMR(400MHz,CDCl3)δ:2.67(m,2H,CH2)；4.05(t,2H,CH2)； 4.56(t,2H,CH2)；5.16(s,2H,CH2)；6.99(s,2H,NH2)；7.38-7.47 (m,5H,C6H5)；7.85-7.84(m,4H,C6H4)；8.09(s,1H,CH)；
ESI-MS:m/z429(M+H)+。
3) Synthesis of (E) -2- (3- (6- (benzyloxy) -2- ((4- (bis (2-hydroxyethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) propyl) isoindoline-1, 3-dione
First, 25mL of a mixture (5mL of acetonitrile, 20mL of dichloromethane, 50uL of trifluoroacetic acid, and 25uL of water) was prepared. At 0 ℃ N2Protected and stirred N9- (3- (N-phthalimido) propyl) -O6To 20mL of a mixture of (1.24g,2.9mmol) of (E) -benzylguanine was added NaNO2(0.76g, 11mmol), and reacted for 25 min. Then, 5mL of a mixture containing sulfamic acid (582mg, 6mmol) was injected and reacted for 20 min. A further 10mL of acetonitrile containing N-phenyldiethanolamine (2g, 12mmol) was injected and reacted at 0 ℃ for 2.5 h. Extracting with dichloromethane and water, mixing organic layers, and extracting with anhydrous Na2SO4Drying overnight, distilling off the solvent under reduced pressure at 30 ℃ and separating and purifying by silica gel column chromatography using acetone and dichloromethane as eluent, gradually changing the volume ratio of acetone/dichloromethane from 1:20 to 1:2 by gradient elution, and drying under vacuum at 30 ℃ to obtain white solid (E) -2- (3- (6- (benzyloxy) -2- ((4- (bis (2-hydroxyethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) propyl) isoindoline-1, 3-dione (1.12g,1.8mmol) with a yield of 62%.
UVλ:226,271nm；
IR (KBr pellet) v/cm-1:3642.1(-OH)；3492.5(N-(CH2)3-)；3362.7 (N-H)；3214.2(C-H)；1841.5(C＝O)；1752.6(N＝N)；1652.7(C＝C)； 1391.5(N-O)；1245.4(C-O-C)；1061.6(C-N)；
1H NMR(400MHz,CDCl3)δ:2.67(m,2H,CH2)；3.61(s,1H,OH)； 3.62(s,1H,OH)；3.73(t,2H,CH2)；4.04(t,2H,CH2)；4.20(t,2H,CH2)；4.56 (t,2H,CH2)；5.18(s,2H,CH2)；6.71-7.75(m,4H,C6H4)；7.35-7.49 (m,5H,C6H5)；7.85-7.89(m,4H,C6H4)；8.08(s,1H,CH)；
ESI-MS:m/z621(M+H)+。
4) Synthesis of (E) -2- (3- (6- (benzyloxy) -2- ((4- (bis (2-chloroethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) propyl) isoindoline-1, 3-dione
To a solution of 1.12g (1.8mmol) of (E) -2- (3- (6- (benzyloxy) -2- ((4- (bis (2-hydroxyethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) propyl) isoindoline-1, 3-dione in 20ml dichloromethane was added 420. mu.L (6.00mmol) SOCl2The reaction was carried out at 30 ℃ for 1.5 hours under reflux. Extracting with dichloromethane and water, mixing organic layers, and extracting with anhydrous Na2SO4Drying overnight at 30 ℃ under reduced pressure to remove the solvent, separating and purifying by silica gel column chromatography using methanol and dichloromethane as eluent, gradually changing the volume ratio of methanol/dichloromethane from 1:1 to 1:4 by gradient elution, and drying under vacuum at 30 ℃ to obtain (E) -2- (3- (6- (benzyloxy) -2- ((4- (bis (2-chloroethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) propyl) isoindoline-1, 3-dione (0.79g,1.2mmol) as a white solid with a yield of 67%.
UVλ:223,272nm；
IR (KBr pellet) v/cm-1:3492.5(N-(CH2)3-)；3362.7(N-H)；3214.2 (C-H)；1841.5(C＝O)；1746.4(N＝N)；1652.7(C＝C)；1391.5(N-O)； 1245.4(C-O-C)；1061.6(C-N)；775.4(C-Cl)；
1H NMR(400MHz,CDCl3)δ:2.65(m,2H,CH2)；3.62(t,2H,CH2)； 3.67(t,2H,CH2)；4.04(t,2H,CH2)；4.57(t,2H,CH2)；5.12(s,2H,CH2)； 6.76-7.74(m,4H,C6H4)；7.32-7.47(m,5H,C6H5)；7.85-7.89(m,4H,C6H4)； 8.05(s,1H,CH)；
ESI-MS:m/z657(M+H)+。
5) Synthesis of (E) -4- ((9- (3-aminopropyl) -6- (benzyloxy) -9H-purin-2-yl) diazenyl) -N, N-bis (2-chloroethyl) aniline
The resulting solid (E) -2- (3- (6- (benzyloxy) -2- ((4- (bis (2-chloroethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) propyl) isoindoline-1, 3-dione (0.79g,1.2mmol) was dissolved in 15mL of DMF and 2.1mL of hydrazine hydrate was added, stirred at 35 ℃ for 5H to allow hydrazinolysis to occur, extracted with dichloromethane and deionized water, the organic phase was washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate overnight, dried under vacuum at 35 ℃ to give (E) -4- ((9- (3-aminopropyl) -6- (benzyloxy) -9H-purin-2-yl) diazenyl) -N, N-bis (2-chloroethyl) aniline (0.47g,0.9mmol), yield 74%.
UVλ:227,278nm；
IR (KBr pellet) v/cm-1:3467.9(N-(CH2)3-)；3347.9(N-H)；3014.3 (C-H)；1802.9(C＝O)；1757.4(N＝N)；1682.1(C＝C)；1417.8(N-O)； 1259.5(C-O-C)；1072.7(C-N)；752.4(C-Cl)；
1H NMR(400MHz,CDCl3)δ:2.54(m,2H,CH2)；2.68(t,2H,CH2)； 3.61(t,2H,CH2)；3.63(t,2H,CH2)；4.05(t,2H,CH2)；5.17(s,2H,NH2)；5.18 (s,2H,CH2)；6.75-7.74(m,4H,C6H4)；7.32-7.49(m,5H,C6H5)；8.12 (s,1H,CH)；
ESI-MS:m/z527(M+H)+。
6) Synthesis of (E) -1- (3- (6- (benzyloxy) -2- ((4- (bis (2-chloroethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) propyl) -3- (2-chloroethyl) urea
The resulting solid (E) -4- ((9- (3-aminopropyl) -6- (benzyloxy) -9H-purin-2-yl) diazenyl) -N, N-bis (2-chloroethyl) aniline (0.47g,0.9mmol) was dissolved in 15mL of dichloromethane, 5mL of dichloromethane containing 2-chloroethyl isocyanate (0.4mL,5mmol) was added dropwise, the reaction was carried out in an ice bath at 0 ℃ for 5 hours, and after completion of the reaction, (E) -1- (3- (6- (benzyloxy) -2- ((4- (bis (2-chloroethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) propyl) -3- (2-chloroethyl) urea (0.38 g) was obtained by distillation under reduced pressure at 28 ℃,0.6mmol), yield 67%.
UVλ:226,274nm；
IR (KBr pellet) v/cm-1:3357.4(N-H)；2997.1(C-H)；1789.3(C＝O)； 1765.4(N＝N)；1676.5(C＝C)；1357.6(N-O)；1268.4(C-O-C)；1146.7 (C-N)；779.8(C-Cl)；
1H NMR(400MHz,CDCl3)δ:2.62(m,2H,CH2)；3.42(t,2H,CH2)； 3.65(t,2H,CH2)；3.66(t,2H,CH2)；3.44(t,2H,CH2)；3.64(t,2H,CH2)； 4.04(t,2H,CH2)；5.16(s,2H,CH2)；6.5(s,1H,NH)；6.76-7.75(m,4H,C6H4)； 7.38-7.47(m,5H,C6H5)；8.11(s,1H,CH)；
ESI-MS:m/z632(M+H)+。
7) Synthesis of (E) -3- (3- (6- (benzyloxy) -2- ((4- (bis (2-chloroethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) propyl) -1- (2-chloroethyl) -1-nitrosourea
Dissolving the obtained solid (E) -1- (3- (6- (benzyloxy) -2- ((4- (bis (2-chloroethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) propyl) -3- (2-chloroethyl) urea (0.38g,0.6mmol) in 6mL acetonitrile, adding 6mL acetonitrile containing glacial acetic acid (5.6. mu.L, 0.1mmol), adding nitrosonium tetrafluoroborate (0.27g,2.3mmol) under ice bath condition, stirring for 5H under ice bath condition, extracting with ethyl acetate and deionized water after reaction, washing the organic phase with saturated sodium chloride aqueous solution, drying with anhydrous sodium sulfate, distilling under reduced pressure to remove the solvent, separating and purifying with silica gel column chromatography, eluting with petroleum ether and ethyl acetate, performing gradient elution, the volume ratio of petroleum ether/ethyl acetate was gradually increased from 1:2 to 1:5, and vacuum drying was carried out at 28 ℃ to give (E) -3- (3- (6- (benzyloxy) -2- ((4- (bis (2-chloroethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) propyl) -1- (2-chloroethyl) -1-nitrosourea (0.23g,0.35mmol) in 58% yield.
UVλ:225,274nm；
IR (KBr pellet) v/cm-1:3389.6(N-H)；3042.5(C-H)；1797.3(C＝O)； 1753.4(N＝N)；1674.5(C＝C)；1589.3(N＝O)；1391.4(N-O)；1281.8 (C-O-C)；1145.9(C-N)；1078.6(N-N)；772.2(C-Cl)；
1H NMR(400MHz,CDCl3)δ:2.58(m,2H,CH2)；3.38(t,2H,CH2)； 3.44(t,2H,CH2)；3.61(t,2H,CH2)；3.63(t,2H,CH2)；3.64(t,2H,CH2)；4.04 (t,2H,CH2)；5.26(s,2H,CH2)；6.1(s,1H,NH)；6.76-7.74(m,4H,C6H4)； 7.38-7.47(m,5H,C6H5)；8.05(s,1H,CH)；
ESI-MS:m/z661(M+H)+。
Example 4: (E) synthesis of (E) -3- (3- (6- (benzyloxy) -2- ((4- (bis (2-chloroethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) propyl) -1- (2-chloroethyl) -1-nitrosourea (Compound 2) N9-bromopropyl-O6Synthesis of-benzylguanine
Weighing O6Adding benzylguanine (1.49g,6.2mmol) and anhydrous potassium carbonate (3.17g,23mmol) into a 50mL round-bottom flask, adding 50mL acetone, slowly heating to 55 ℃, dropwise adding 1, 3-dibromopropane (3.33mL,30mmol), continuing to react for 72 hours after the dropwise addition is finished, filtering the reaction solution, collecting the filtrate, removing the solvent by reduced pressure distillation at 45 ℃, separating and purifying by silica gel column chromatography, using petroleum ether and ethyl acetate as eluent, gradually increasing the volume ratio of the petroleum ether to the ethyl acetate from 1:2 to 1:4 by gradient elution, and drying in vacuum at 40 ℃ to obtain a white solid N9-bromopropyl-O6Benzylguanine (1.48g,4.1mmol), 66% yield.
UVλ:250,284nm；
IR (KBr pellet) v/cm-1:3456.4(N-(CH2)3-)；3318.2(N-H)；2950.2 (C-H)；1642.2(C＝C)；1251.2(C-O-C)；1055.9(C-N)；742.2(C-Br)；
1H NMR(400MHz,CDCl3)δ:2.22(m,2H,CH2)；3.54(t,2H,CH2)； 4.09(t,2H,CH2)；5.16(s,2H,CH2)；6.97(s,2H,NH2)；7.35-7.41 (m,5H,C6H5)；8.14(s,1H,CH)；
ESI-MS:m/z362(M+H)+。
2) N9- (3- (N-phthalimido) propyl) -O6Synthesis of-benzylguanine
The solid obtained was N9-bromopropyl-O6-benzylguanine (1.48g,4.1mmol), dissolving in 15mL of anhydrous N, N-dimethylformamide, adding solid potassium phthalimide (1.66g,9mmol), heating to 65 deg.C, stirring for reaction for 6h, extracting with ethyl acetate and deionized water, collecting the organic phaseWashing with saturated aqueous sodium chloride solution, drying over night with anhydrous sodium sulfate, and vacuum drying at 40 deg.C to obtain white solid N9- (3- (N-phthalimido) propyl) -O6Benzylguanine (1.34g,3.12mmol), yield 76%.
UVλ:225,286nm；
IR (KBr pellet) v/cm-1:3478.2(N-(CH2)3-)；3339.2(N-H)；2971.4 (C-H)；1785.4(C＝O)；1634.2(C＝C)；1283.2(C-O-C)；1092.2(C-N)；
1H NMR(400MHz,CDCl3)δ:2.65(m,2H,CH2)；4.08(t,2H,CH2)； 4.59(t,2H,CH2)；5.17(s,2H,CH2)；6.72(s,2H,NH2)；7.36-7.45 (m,5H,C6H5)；7.81-7.86(m,4H,C6H4)；8.11(s,1H,CH)；
ESI-MS:m/z429(M+H)+。
3) Synthesis of (E) -2- (3- (6- (benzyloxy) -2- ((4- (bis (2-hydroxyethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) propyl) isoindoline-1, 3-dione
First, 25mL of a mixture (5mL of acetonitrile, 20mL of dichloromethane, 50uL of trifluoroacetic acid, and 25uL of water) was prepared. At 0 ℃ N2Protected and stirred N9- (3- (N-phthalimido) propyl) -O6To 20mL of a mixture of (1.34g,3.12mmol) of (E) -benzylguanine was added NaNO2(0.75g, 10.92mmol), and reacted for 25 min. Then, 5mL of a mixed solution containing sulfamic acid (601mg, 6.2mmol) was injected and reacted for 20 min. A further 10mL of acetonitrile containing N-phenyldiethanolamine (2.15g, 12.8mmol) was injected and reacted at 0 ℃ for 2.5 h. Extracting with dichloromethane and water, mixing organic layers, and extracting with anhydrous Na2SO4Drying overnight at 30 deg.C under reduced pressure to remove solvent, separating and purifying by silica gel column chromatography using acetone and dichloromethane as eluent, gradient elution was performed, the volume ratio of acetone/dichloromethane was gradually changed from 1:20 to 1:2, and vacuum drying at 30 deg.C gave (E) -2- (3- (6- (benzyloxy) -2- ((4- (bis (2-hydroxyethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) propyl) isoindoline-1, 3-dione (1.24g,2mmol) as a white solid in 64% yield.
UVλ:226,271nm；
IR (KBr pellet) v/cm-1:3649.1(-OH)；3496.1(N-(CH2)3-)；3358.3 (N-H)；3217.3(C-H)；1847.1(C＝O)；1755.1(N＝N)；1659.1(C＝C)； 1390.2(N-O)；1246.2(C-O-C)；1065.2(C-N)；
1H NMR(400MHz,CDCl3)δ:2.68(m,2H,CH2)；3.60(s,1H,OH)； 3.65(s,1H,OH)；3.75(t,2H,CH2)；4.07(t,2H,CH2)；4.23(t,2H,CH2)；4.58 (t,2H,CH2)；5.17(s,2H,CH2)；6.68-7.74(m,4H,C6H4)；7.35-7.45 (m,5H,C6H5)；7.81-7.89(m,4H,C6H4)；8.14(s,1H,CH)；
ESI-MS:m/z621(M+H)+。
4) Synthesis of (E) -2- (3- (6- (benzyloxy) -2- ((4- (bis (2-chloroethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) propyl) isoindoline-1, 3-dione
To a solution of 1.24g (2mmol) of (E) -2- (3- (6- (benzyloxy) -2- ((4- (bis (2-hydroxyethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) propyl) isoindoline-1, 3-dione in 20mL dichloromethane was added SOCl2(560. mu.L, 8.00mmol) under reflux at 30 ℃ for 1.5 hours. Extracting with dichloromethane and water, mixing organic layers, and extracting with anhydrous Na2SO4Drying overnight, distilling off the solvent under reduced pressure at 30 ℃ and separating and purifying by silica gel column chromatography using methanol and dichloromethane as eluent, gradually changing the volume ratio of methanol/dichloromethane from 1:1 to 1:4 by gradient elution, and drying under vacuum at 30 ℃ to obtain white solid (E) -2- (3- (6- (benzyloxy) -2- ((4- (bis (2-chloroethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) propyl) isoindoline-1, 3-dione (0.92g,1.4mmol) with a yield of 70%.
UVλ:223,272nm；
IR (KBr pellet) v/cm-1:3495.2(N-(CH2)3-)；3365.2(N-H)；3218.1 (C-H)；1845.2(C＝O)；1756.3(N＝N)；1662.3(C＝C)；1385.5(N-O)； 1253.9(C-O-C)；1065.2(C-N)；781.6(C-Cl)；
1H NMR(400MHz,CDCl3)δ:2.66(m,2H,CH2)；3.64(t,2H,CH2)； 3.62(t,2H,CH2)；4.09(t,2H,CH2)；4.55(t,2H,CH2)；5.13(s,2H,CH2)； 6.75-7.74(m,4H,C6H4)；7.32-7.45(m,5H,C6H5)；7.85-7.89(m,4H,C6H4)；； 8.13(s,1H,CH)；
ESI-MS:m/z657(M+H)+。
5) (E) -4- ((9- (3-aminopropyl) -6- (benzyloxy) -9H-purin-2-yl) diazenyl) -N,
synthesis of N-bis (2-chloroethyl) aniline
The resulting solid (E) -2- (3- (6- (benzyloxy) -2- ((4- (bis (2-chloroethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) propyl) isoindoline-1, 3-dione (0.92g,1.4mmol) was dissolved in 15mL of DMF and 2.5mL of hydrazine hydrate was added, the reaction was stirred at 35 ℃ for 6H to allow hydrazinolysis, extraction was performed with dichloromethane and deionized water, the organic phase was washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate overnight, dried under vacuum at 35 ℃ to give (E) -4- ((9- (3-aminopropyl) -6- (benzyloxy) -9H-purin-2-yl) diazenyl) -N, N-bis (2-chloroethyl) aniline (0.58g,1.1mmol), yield 79%.
UVλ:227,278nm；
IR (KBr pellet) v/cm-1:3473.4(N-(CH2)3-)；3352.6(N-H)；3011.2 (C-H)；1803.4(C＝O)；1751.1(N＝N)；1678.6(C＝C)；1419.2(N-O)； 1242.5(C-O-C)；1075.72(C-N)；753.4(C-Cl)；
1H NMR(400MHz,CDCl3)δ:2.56(m,2H,CH2)；2.72(t,2H,CH2)； 3.63(t,2H,CH2)；3.68(t,2H,CH2)；4.06(t,2H,CH2)；5.12(s,2H,NH2)；5.16 (s,2H,CH2)；6.75-7.76(m,4H,C6H4)；7.32-7.45(m,5H,C6H5)；8.15 (s,1H,CH)；
ESI-MS:m/z527(M+H)+。
6) Synthesis of (E) -1- (3- (6- (benzyloxy) -2- ((4- (bis (2-chloroethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) propyl) -3- (2-chloroethyl) urea
The resulting solid (E) -4- ((9- (3-aminopropyl) -6- (benzyloxy) -9H-purin-2-yl) diazenyl) -N, N-bis (2-chloroethyl) aniline (0.58g,1.1mmol) was dissolved in 15mL of dichloromethane, 6mL of dichloromethane containing 2-chloroethyl isocyanate (0.48mL,6mmol) was added dropwise, the reaction was carried out in an ice bath at 0 ℃ for 6 hours, and after completion of the reaction, (E) -1- (3- (6- (benzyloxy) -2- ((4- (bis (2-chloroethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) propyl) -3- (2-chloroethyl) urea (0.47 g) was obtained by distillation under reduced pressure at 28 ℃,0.75mmol), yield 68%.
UVλ:226,274nm；
IR (KBr pellet) v/cm-1:3362.2(N-H)；2985.3(C-H)；1792.3(C＝O)； 1773.4(N＝N)；1761.5(C＝C)；1363.2(N-O)；1262.4(C-O-C)；1145.2 (C-N)；779.8(C-Cl)；
1H NMR(400MHz,CDCl3)δ:2.61(m,2H,CH2)；3.45(t,2H,CH2)； 3.62(t,2H,CH2)；3.69(t,2H,CH2)；4.04(t,2H,CH2)；5.17(s,2H,CH2)；6.5 (s,1H,NH)；6.76-7.79(m,4H,C6H4)；7.38-7.45(m,5H,C6H5)；8.14(s,1H,CH)；
ESI-MS:m/z632(M+H)+。
7) Synthesis of (E) -3- (3- (6- (benzyloxy) -2- ((4- (bis (2-chloroethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) propyl) -1- (2-chloroethyl) -1-nitrosourea
Dissolving the obtained solid (E) -1- (3- (6- (benzyloxy) -2- ((4- (bis (2-chloroethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) propyl) -3- (2-chloroethyl) urea (0.47g,0.75mmol) in 5mL acetonitrile, adding 5mL acetonitrile containing glacial acetic acid (11.2. mu.L, 0.2mmol), adding nitrosonium tetrafluoroborate (0.33g,2.8mmol) under ice bath condition, stirring for 5H under ice bath condition, extracting with ethyl acetate and deionized water after reaction, washing the organic phase with saturated sodium chloride aqueous solution, drying with anhydrous sodium sulfate, distilling under reduced pressure to remove the solvent, separating and purifying with silica gel column chromatography, eluting with petroleum ether and ethyl acetate, performing gradient elution, the volume ratio of petroleum ether/ethyl acetate was gradually increased from 1:2 to 1:5, and vacuum drying was carried out at 28 ℃ to give (E) -3- (3- (6- (benzyloxy) -2- ((4- (bis (2-chloroethyl) amino) phenyl) diazenyl) -9H-purin-9-yl) propyl) -1- (2-chloroethyl) -1-nitrosourea (0.3g,0.45mmol) in 61% yield.
UVλ:225,274nm；
IR (KBr pellet) v/cm-1:3383.6(N-H)；3041.2(C-H)；1788.2(C＝O)； 1756.5(N＝N)；1680.2(C＝C)；1593.1(N＝O)；1392.4(N-O)；1289.2 (C-O-C)；1146.9(C-N)；1075.6(N-N)；771.9(C-Cl)；
1H NMR(400MHz,CDCl3)δ:2.57(m,2H,CH2)；3.36(t,2H,CH2)； 3.43(t,2H,CH2)；3.60(t,2H,CH2)；3.63(t,2H,CH2)；3.69(t,2H,CH2)；4.02 (t,2H,CH2)；5.25(s,2H,CH2)；6.13(s,1H,NH)；6.72-7.77(m,4H,C6H4)； 7.33-7.41(m,5H,C6H5)；8.13(s,1H,CH)；
ESI-MS:m/z661(M+H)+。
The novel chloroethyl nitrosourea compound prepared by the invention has the following evaluation on the anti-tumor activity: experimental example 1: evaluation of antitumor Activity
1. Experimental materials and instruments
Test compounds: compound 1 and compound 2 prepared in the above preparation examples;
cell line: l1210 mouse leukemia cell, HT29 human colon cancer cell, A549 human lung cancer cell, HUT102 human T lymphoma cell, human glioma cell SF763, SF126
2. Experimental methods
The six tumor cells were inoculated into a 96-well plate at 1000/well and 5% CO at 37 deg.C2After 24 hours of incubation, 5 replicates of each group were replaced with carmustine (positive control), compound 1 and compound 2 at serial concentrations (1 μ M, 5 μ M, 10 μ M, 50 μ M, 100 μ M, 200 μ M, 400 μ M and 1000 μ M) and the control group was set. The above groups were exposed to aerobic and hypoxic conditions for 48 hours, respectively. Then, 10. mu.L of CCK-8 solution was added to each well and allowed to act for 4 hours. Finally, the absorbance value at 450nm was measured, and the cell activity was calculated by the following formulaCalculating by regression analysis to obtain median inhibition rate IC50。
Cell survival rate (%) ═ aMedicine adding device–ABlank group)/(AControl group–ABlank group)×100
AMedicine adding deviceAbsorbance of wells with cells, CCK8 solution, and drug solution;
Ablank groupAbsorbance for wells with medium and CCK8 solution without cells;
control a is the absorbance of wells with cells, CCK8 solution, and no drug solution.
3. The experimental results are as follows: see Table 1
TABLE 1 half inhibition rate (IC) of tumor cells50,μM)
Table 1 the results show that:
IC of Compounds 1 and 2 on 6 tumor cells in normoxic Environment50The values were similar to those of the positive control group (carmustine group), indicating that the tumor cell inhibitory activity of compounds 1 and 2 was not much different from that of the carmustine group under normoxic conditions.
IC of Compounds 1 and 2 on 6 tumor cells in hypoxic Environment50The values were significantly lower than the positive control group (carmustine group). This indicates that compounds 1 and 2 are capable of liberating O on the one hand by breaking the azo bond under low oxygen conditions6-benzylguanine analogues as AGT inhibitors, thereby blocking AGT mediated resistance of tumor cells and rendering cancer cells more susceptible to nitrosourea alkylation; on the other hand, the compounds 1 and 2 are subjected to the breakage of azo bonds under the hypoxic environment, so that aromatic nitrogen mustard is released and is used as another alkylating agent to attack the N7 position of guanine, and the DNA of cancer cells generates intrachain or interchain crosslinking. Thereby leading the antitumor activity of the compounds 1 and 2 to be obviously improved.
IC comparing Compounds 1 and 2 in normoxic and hypoxic environments50Values, it can be seen that the tumor cell inhibition of Compounds 1 and 2 is lower in hypoxic than in normoxic environmentsThe activity is obviously improved, and the compounds 1 and 2 have low oxygen environment selectivity. Therefore, the compounds 1 and 2 can selectively act on tumor cells in a hypoxic state, and avoid damage to normal cells in an normoxic state, so that the aim of targeting on the tumor cells is fulfilled.
The experimental result shows that the compound provided by the invention has higher tumor inhibition activity than the prior chloroethylnitrosourea under the low oxygen condition; meanwhile, the compounds can specifically act on tumor cells and can be used for tumor targeted therapy.
Although the invention has been described in detail hereinabove by way of general description, specific embodiments and experiments, it will be apparent to those skilled in the art that many modifications and improvements can be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (12)
1. A compound or pharmaceutically acceptable salt having the structure of formula (I):
r is H, NH2，(CH2)nNH2，CH3，CH2CH3，CH2CH2CH3，CH2OH，CH2CH2OH，NHCOCH3，(CH2)nNHCOCH3，N(CH3)2，(CH2)nN(CH3)2One of (1);
n is an integer of 2 to 6.
2. The compound or pharmaceutically acceptable salt of claim 1, wherein R is H, NH2，N(CH3)2Or NHCOCH3。
3. The compound or pharmaceutically acceptable salt according to claim 1, wherein the compound is selected from one of the compounds of the following structures:
4. the compound or pharmaceutically acceptable salt according to any one of claims 1 to 3, wherein the pharmaceutically acceptable salt is a hydrochloride, hydrobromide, hydroiodide, sulfate, bisulfate, phosphate, acetate, propionate, butyrate, lactate, methanesulfonate, p-toluenesulfonate, maleate, benzoate, succinate, tartrate, citrate, fumarate, taurate or gluconate salt.
5. A pharmaceutical composition characterized by: the pharmaceutical composition comprises an effective amount of a compound or pharmaceutically acceptable salt of any one of claims 1 to 4, and at least one pharmaceutically acceptable carrier.
6. The pharmaceutical composition of claim 5, wherein the pharmaceutical composition is an injection, a tablet or a capsule.
7. A process for the preparation of a compound or pharmaceutically acceptable salt according to any one of claims 1 to 4, wherein the reaction sequence comprises:
(1) reacting the compound a with dibromoalkane or dichloroalkane to obtain a compound b;
(2) reacting the compound b with potassium phthalimide to obtain a compound c;
(3) sequentially adding NaNO into the compound c2Reacting sulfamic acid and N, N-dihydroxyethylaniline to obtain a compound d;
(4) reacting the compound d with thionyl chloride to obtain a compound e;
(5) reacting the compound e with hydrazine hydrate to obtain a compound f;
(6) reacting the compound f with 2-chloroethyl isocyanate to obtain a compound g;
(7) reacting the compound g with nitrosonium tetrafluoroborate to obtain a compound h;
the R or n is selected from the R or n of any one of claims 1 to 4, the R1Is Cl or Br.
8. The method according to claim 7, characterized in that it comprises the steps of:
(1) compound a is reacted with dibromoalkane or dichloroalkane according to a ratio of 1: (1-7) reacting at 40-60 ℃ under the catalysis of alkali to obtain a compound b;
(2) compound b was reacted with potassium phthalimide according to 1: (1-4) reacting at 35-70 ℃ to obtain a compound c;
(3) according to the compound c, NaNO2The mol ratio of the sulfamic acid to the N, N-dihydroxyethylaniline is 1: (1-4): (1-5): (3-10) adding NaNO to the Compound c2In N at2Reacting for 0-1h at 0-10 ℃ under protection, adding sulfamic acid, reacting for 0-1h at 0-10 ℃, finally adding N, N-dihydroxyethylaniline, and reacting for 1-4h at 0-10 ℃ to obtain a compound d;
(4) compound d with SOCl2According to the following steps of 1: (1-7) carrying out reflux reaction at the temperature of 20-40 ℃ to obtain a compound e;
(5) reacting the compound e with hydrazine hydrate at the temperature of 25-60 ℃ to obtain a compound f;
(6) compound f with 2-chloroethyl isocyanate according to 1: (1-8) reacting at 0-10 ℃ to obtain a compound g;
(7) compound g was reacted with nitrosonium tetrafluoroborate according to 1: (1-5) reacting at 0-10 ℃ to obtain a compound h.
9. The method according to claim 8, wherein in the step (1), the compound a is reacted with dibromoalkane.
10. The compound or pharmaceutically acceptable salt according to any one of claims 1 to 4, and the pharmaceutical composition according to claim 5, for use in the preparation of an antitumor drug.
11. Use according to claim 10, characterized in that: the tumor is one or more of brain tumor, myeloma, malignant melanoma, malignant lymphoma, lung cancer, breast cancer, gastric cancer, colon cancer, prostate tumor, leukemia, and lymph cancer.
12. The use according to claim 11, wherein the tumor is one or more of brain tumor, leukemia, lung cancer, colon cancer, and lymphoma.
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| CN104031047A (en) * | 2014-05-28 | 2014-09-10 | 北京工业大学 | Novel chloroethyl nitrosourea with anticancer activity and synthesizing method thereof |
| CN104031048A (en) * | 2014-05-28 | 2014-09-10 | 北京工业大学 | Novel beta-chloroethylnitrosourea compounds, and synthesis method and application thereof |
| CN105503874A (en) * | 2015-12-29 | 2016-04-20 | 北京工业大学 | Low-oxygen activated joint chloroethane nitrourea compound and preparation method and application |
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| CN104031048A (en) * | 2014-05-28 | 2014-09-10 | 北京工业大学 | Novel beta-chloroethylnitrosourea compounds, and synthesis method and application thereof |
| CN105503874A (en) * | 2015-12-29 | 2016-04-20 | 北京工业大学 | Low-oxygen activated joint chloroethane nitrourea compound and preparation method and application |
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