WO2011082519A1

WO2011082519A1 - Geldanamycin derivatives, preparation methods and uses thereof

Info

Publication number: WO2011082519A1
Application number: PCT/CN2010/002196
Authority: WO
Inventors: 林国强; 冯陈国
Original assignee: 杭州华东医药集团生物工程研究所有限公司; 上海任洲生化科技有限公司
Priority date: 2010-01-05
Filing date: 2010-12-29
Publication date: 2011-07-14
Also published as: CN102115460A; CN102115460B

Abstract

Geldanamycin derivatives, preparation methods and uses thereof in the manufacture of medicaments of cell killing agents, cell proliferation inhibitors and antitumor agents are disclosed.

Description

Geldanamycin derivative, preparation method and use thereof

The present invention relates to a novel class of geldanamycin derivatives, a process for preparing such compounds, and the use of such compounds for the preparation of cell killing activity cell killing agents, cell proliferation inhibitors and antitumor drugs. Background technique

Geldanamycin was first discovered in 1970 from the Streptomyces product [C. DeBoer, et al.; Geldanamycin, a new antibiotic: J. Antibiot" 1970, 23(9), 442-447], and thereafter, from Many similar compounds have been discovered in microbial products or by artificial synthesis and have been found to have a variety of biological activities. For example, [M. Muroi, et α/·; The structures of macbecin I and II: Tetrahedron, 1981, 37, Pp. 1123-1131], literature [RC Schnur, et al.; Inhibition of the oncogene product ^lS5 ^erbB " ² in vitro and in vivo by geldanamycin and dihydrogeldanamycin derivatives: J. Med. Che , 1995, 38, 3806-3812 】, literature [Μ· Bendin, et al. Geldanamycin, an inhibitor of the chaperone activity of HS90, induces MAPK-independent cell cycle arrest: Int. J. Cancer, 2004, 109, 643-652], literature [Z.- Q. Tian et air, Synthesis and biological activities of novel 17-aminogeldanamycin derivatives: Bioorg. Med. Chem" 2004, 12, 5317-5329] and literature [J.-YL Brazidec, et aL; Synthesis and biological evaluation of a new Class of geldanam Ycin derivatives as potent inhibitors of Hsp90: J. Med. Chenu, 2004, 47, 3865-3873] have described natural or synthetic geldanamycins and their biological activities. More associated with heat shock protein 90. Heat shock protein 90 is the most active molecular chaperone in the cell, many signaling pathways are dependent on heat shock eggs White 90, and its expression in tumor cells is 2 to 10 times higher than that of normal cells, and may play an important regulatory role in tumor cell growth and survival. Geldanamycins can specifically bind to heat shock protein 90 and inhibit their function, leading to the degradation of various oncogene products and cell cycle regulatory proteins, thereby showing various biological activities such as anticancer, and therefore, such compounds It has received great attention and in-depth research by cancer researchers. Among them, 17-allylamine-17-demethoxygeldanamycin (17-AAG) is undergoing clinical trials for the treatment of tumors in the United States.

The geldanamycin derivative of the present invention belongs to an ansamycin antibiotic, and structurally, all of the geldanamycin compounds which have been described so far have not been observed to have the same structure as the compound of the present invention. Similar compounds. Summary of the invention

The present invention aims to provide a geldanamycin derivative having cell killing activity, cell proliferation inhibiting activity and antitumor activity.

The inventors have found through hard work that a class of geldanamycin derivatives having cell killing activity, cell proliferation inhibitory activity and antitumor activity are selected from the following formulas (1) to (14):

Formula 1 )

-3-

Formula (7)

Sa US OAV

Formula (13)

In the present invention, for the convenience of description, the structure described in the following structural formula (15) is defined as GA, that is,

' X 0

(15)

The modifications described in the present invention are all carried out at the 17 position in the formula (15). The present invention also provides a method of synthesizing the geldanamycin derivatives represented by the formulae (1) to (14).

The separation and purification includes conventional methods of separation and purification using natural products well known to those skilled in the art, such as liquid-liquid extraction, column chromatography, thin layer chromatography, and recrystallization.

The present invention also provides a method of inhibiting cell growth, inhibiting cell proliferation, or treating a tumor, comprising administering to a subject a therapeutically effective amount of any geldanamycin derivative represented by the chemical formulas (1) to (14). The step of the substance or its salt. The term "pharmaceutically acceptable salt" in the present invention may be a pharmaceutically acceptable inorganic or organic salt. The compound having a basic group in the formula I of the present invention may form a pharmaceutically acceptable salt with a mineral acid, such as a sulfate or a hydrochloric acid. Salts, hydrobromides, phosphates; also form pharmaceutically acceptable salts with organic acids, such as acetates, oxalates, citrates, gluconates, succinates, tartrates, p-toluenesulfonates , mesylate, benzoate, lactate, maleate, etc. The compound having an acidic group in the formula I of the present invention may form a pharmaceutically acceptable salt with an alkali metal or an alkaline earth metal, preferably but not limited to a sodium salt. , potassium, magnesium or calcium salts.

The compounds of formula I of the present invention can be combined with various pharmaceutically acceptable carriers, excipients or adjuvants to form antitumor agents for the treatment of tumors.

The compounds of the invention may be administered alone or in the form of a pharmaceutical composition. The route of administration may be oral, parenteral or topical. The pharmaceutical composition can be formulated into various suitable dosage forms depending on the route of administration.

The pharmaceutical composition of the compound of the present invention can be administered in any of the following ways: oral, spray inhalation, rectal administration, nasal administration, buccal administration, topical administration, parenteral administration, such as subcutaneous, intravenous, intramuscular, intraperitoneal, sheath Inside, intraventricular, intrasternal and intracranial injection or input, or by means of an explant reservoir. Among them, oral, intraperitoneal or intravenous administration is preferred.

When administered orally, the compounds of the invention may be formulated in any orally acceptable form including, but not limited to, tablets, capsules, aqueous solutions or aqueous suspensions. Among them, the carrier used for the tablet generally comprises lactose and corn starch, and a lubricant such as magnesium stearate may also be added. The diluent used in the capsule preparation generally comprises lactose and dried corn starch. Aqueous suspension formulations are usually prepared by admixing the active ingredient with a suitable emulsifier or suspension. Optionally, some of the above oral preparation forms may also contain some sweeteners, fragrances or colorants.

When the skin is applied topically, the compounds of the invention may be formulated in the form of a suitable ointment, lotion or cream preparation in which the active ingredient is suspended or dissolved in one or more carriers. Carriers for ointment preparations include, but are not limited to: mineral oil, liquid petroleum jelly, White petrolatum, propylene glycol, polyethylene oxide, polypropylene oxide, emulsifying wax and water; detergents or creams can be used, including but not limited to: mineral oil, sorbitan monostearate, Tween 60, ten Hexadecane wax, hexadecene aromatic alcohol, 2-octyldodecanol, benzyl alcohol and water.

The compounds of the present invention can also be administered in the form of a sterile injectable preparation, including sterile aqueous or oily suspension or sterile injection solutions. Among them, carriers and solvents which can be used include water, Ringer's solution and isotonic sodium chloride solution. Alternatively, sterilized, fixed oils may be employed as a solvent or suspension medium such as a monoglyceride or a diglyceride.

It should also be noted that the dosage and method of use of the compounds of the invention depend on a number of factors, including the age, weight, sex, natural health, nutritional status of the compound, the strength of the compound, the time of administration, the rate of metabolism, the severity of the condition, and the diagnosis and treatment. Subjective judgment of the physician. The preferred dosage is between 0.01 and 100 mg/kg body weight per day.

The compounds of the formula (1) - (14) of the present invention can also be used as a low molecular biological probe for inhibiting cell cycle in life science research. When the compound represented by the formula (1) - (14) is used in life science research, it is soluble in methanol or aqueous sterol, and is also soluble in an aqueous solution of dimethyl sulfoxide. detailed description

The following examples are intended to further illustrate the invention but are not to be construed as limiting. Test Materials: Nuclear magnetic resonance was measured by a Varian EM-360A, EM-390 or Bruker AMX-300 instrument. Mass spectra were determined by a Finnigan 4021, HP5989A, Finnigan FTMS-2000 instrument. Flash column chromatography was performed on silica gel (300-400 mesh).

TLC was assayed with HSGF254 high efficiency plates using UV lamp 254 and 365 nm wavelength or 5% phosphomolybdate ethanol solution. The completion endpoint of the reaction was determined by TLC assay. The reagents used in the experiments were all commercial reagents and were used without purification. The amine fragments used in the experiment were partially purchased through commercialization, and could not be purchased by the following two synthetic routes. Get it,

Comparative example 1

In parallel comparison with the 17 modified geldanamycin derivatives entering the clinical study, Applicants synthesized the two comparative compounds 1 and 2 shown in Table 1 for comparison purposes: RZC01 and RZC02. Synthetic methods reference (Shen, Y., Xie, Q., Norberg, Μ·, et al. Bioorg. Med. Chem. 2005, 13, 4960.).

Table 1

Comparative compound name structural formula molecular weight

1

RZC02, N A 617

z NH-GA

2

RZC03 ^{3⁄43⁄43⁄4/} ^ H-GA 568

Example 1 Add dopamine hydrochloride (1.35 g) to geldanamycin (2 g), then add dichloromethane (100 mL), triethylamine (1.5 mL), methanol (33 mL), stir at room temperature reaction. The reaction formula is as follows:

The reaction system was initially orange-red and gradually darkened to black. The reaction was completed for 2.5 days. The solvent was evaporated under reduced pressure and purified (EtOAcjjjjjjjjjjjj The product was subjected to analytical testing to obtain the following spectral data:

^{_{! H NMR (CDC1 3, 300}} MHz) δ 0.94 (d, J = 5.7 Hz, 3H); 1.00 (d, J = 6.6 Hz, 3H); 1.62-1.84 (obscure, m, 8H); 2.02 (s, 3H); 2.32-2.48 (m, IH); 2.58-2.90 (m, 4H); 3.28 (s, 3H); 3.37 (s, 3H); 3.35-3.47 (m 2H), 3.48-3.82 (m, 3H ); 4.33 (d, br, IH); 4.54 (s, br, IH); 4.90 (s, br, 2H); 5.19 (s, IH); 5.82-5.98 (m, 2H); 6.16 (br, s , IH); 6.38-6.47 (m, IH); 6.52-6.64 (m, 2H); 6.68 -6.75 (m, 2H); 6.83 (br, d, J = 8.1 Hz, IH); 6.95 (br, d , J = 11.7 Hz, IH); 7.22 (s, IH); 9.17 (s, 1H).

¹³ C NMR ( acetone- , 75 MHz ) δ 12.46, 12.90, 13.14, 23.22, 29.07, 33.13, 34.70, 35.95, 47.77, 56.68, 56.74, 73.20, 81.59, 82.20, 82.27, 108.46, 109.26, 116.26, 116.60, 120.97 , 126.91, 128.80, 130.64, 133.44, 134.27, 134.87, 137.71, 138.69, 144.70, 145.96, 146.11, 157.12, 169.39, 180.79, 184.82.

ESI (m/z): 704.2 (M + Na ⁺ ).

It is proved that the product of the formula (1) is obtained: -z\ -

°9L'£Sl '8SO8I '€Κ89Τ -9V9SI 'SSl 'ZL't l -LVi -ZL'Sil -Setil -S9'££l -ZL'Z£l '6L'6l\ 'W^Zl 88· 9ΖΪ '099^1 i£8SIl ^69*801 ^S*80l -6£'IS 'tl'lS '99'ZL -OI LS -69'9S -60'LP -ΖΟ'ξϊ -6£'t£ -SZ'Z£ -OS'SZ -LV Z i98 Z -SL'Zl -1911 -L£'Zl 9 (ZHPM SL '£1303 ) WL 3 _EI 8 (HI 's) LV6 'ZH L'S = f 'Ρ SZ'L KHZ '^Η Γ8 = f 'P) LO'L HT 'ZH pu = r 'jq ⁴ p) HZ 'ZH = r 'P)T8*9 KHI 'ZH in = r 8S*9 Km 6 £ 9-τε·9 KHI 'S) ei*9 KHZ; ^6 s-i8 s HI 'S) 6i s KHZ; '« 'jq) β ρ -(HI 'S 'jq) KHI 'ZH 66 = r 'p) i£ t '∞) es'e-^s'e KHI '« os*e-ore HC 'S) 9ε·ε

's) Are HZ '« ε6 -ζ8*ζ -(HZ IS -Z9 '« 8^ -9ε

KH£ 'S) ZO'Z H9 'ajnosqo) £8 I"T9 TH£ 'ZH 6*9 = f 'Ρ) 001 - ( H£ 'ZH 99 = f ' ) S609 ( z brain 00€ ' ^C 13QD ) H _t

' ^ . (%8l 'Sui £ ) Z08ZH 3⁄4 '^g

( Ί"ΐ S*0 ) ^ ' (Ί«ΙΖΪ) 3⁄4 ' (im OS)

f-Tfl l ί ^

96TZ00/0T0ZN3/X3d 6TSZ80/H0Z: OAV ESI (m/z): 688.3 (M + Na).

It is proved that the product of the formula (2) is obtained:

Formula (2)

Example 3

Geldanamycin (1 g) was added, [(2-amino)-ethylthiomethylfuran (0.5 mL) was added, dichloromethane (100 mL) was added, and the reaction was stirred at room temperature, and the system was an orange-red suspension. The reaction formula is as follows:

The reaction was stopped for 8 days. The solvent was evaporated under reduced pressure and purified by column chromatography (eluent: methylene chloride / decyl alcohol = 40:1), and then purified by preparative TLC to give a purple solid, which was named RZ803 (150 mg, 12%). The product was subjected to analytical testing to obtain the following spectral data:

^{_{! H NMR (CDC1 3, 300}} MHz) δ 0.90-1.10 (m, 6H); 1.62-1.90 (obscure, m, 3H); 1.80 (s, 3H); 2.03 (s, 3H); 2.26-2.37 (m , 1H); 2.62-2.87 (m, 4H); 3.72 (d, J = 2.1 Hz, 3H); 3.37 (d, J = 0.9 Hz, 3H); 3.42-3.70 (m, 4H); 3.74 (s, 3H); 4.26-4.40 (m, 1H); 4.94 (br, s, 2H); 5.19 (br, s, 1H); 5.81-5.96 (m, 2H); 6.22 (br, s, 1H); 6.50- 6.65 (m, 2H); 6.94 (br, d, J = 11.4 Hz, lH); 7.29 (d, J = 2.4 Hz, 1H); 7.38 (s, 1H); 9.14 (s, 1H).

¹³ C NMR ( CDCI 3, 75 MHz ) δ 12.34; 12.60; 12.78; 22.95; 27.86; 28.55; 31.10; 32.26; 34.41; 35.00; 43.92; 56.71; 57.10; 72.55; 81.14; 81.37; 81.61; 107.94; 108.90; 109.09; 110.61; 126.55; 126.90; 132.76; 133.69; 134.94; 135.79; 141.03; 142.45; 144.68; 150.86; 156.03; 168.38; 180.78;

ESI (m/z): 708.1 (M + Na+). It is proved that the product of the formula (3) is obtained:

Formula ( ₃ )

Example 4

Geldanamycin (567 mg), 2,2-diphenyl-1-propylamine (496 mg), dichloromethane (50 mL), methanol (12 mL), triethylamine (0.45 mL), stir at room temperature

The reaction was stopped for 6 days. The solvent was evaporated under reduced pressure and purified by silica gel column chromatography (eluent: methylene chloride/methanol = 20:1). The product was subjected to analytical testing to obtain spectral data as follows:

lB匪R ( CDC1 ₃ , 300 MHz ) δ 0.96 (d, J = 6.3 Hz, 3H); 1.01 (d, J = 6.9 Hz, 3H); 1.62-1.95 (m, 9H); 2.02 (s, 3H) 2.32-2.48 (m, IH) 2.63-2.80 (m, 2H); 3.26 (s, 3H); 3.37 (s, 3H); 3.42-3.64 (m, 2H) 3.96-4.05 (m, IH); 4.21 -4.46 (m, 3H); 4.90 (s, br, 2H); 5.19 (s, IH) 5.81-5.94 (m, 2H); 6.19 (s, br, IH); 6.58 (t, J = 11.4 Hz, IH); 6.94 (d, br, J = 11.7 Hz, IH); 7.15-7.41 (m, 11H); 9.14 (s, 1H).

¹ C NMR ( CDC1 ₃ , 75 MHz ) δ 12.41; 12.59; 12.79; 23.07; 27.03 28.47; 32.27; 34.67; 35.07; 47.55; 55.37; 56.69; 57.11; 72.54; 81.12 81.41; 81.60; 108.78; 126.55; 126.87; 127.01; 127.23; 127.24 128.75; 132.71; 133.67; 134.93; 135.76; 141.10; 145.18; 145.34 145.35; 156.01; 168.38; 180.54;

ESI (m/z): 726.3 (M + Na+).

It is proved that the product of the formula (4) is obtained: NH-GA

P formula ( 4 )

Example 5

Geldanamycin (561 mg), adding 3-(diethoxymethylsilyl)-1-propanamine (840 mg), adding dichloromethane (50 mL), stirring at room temperature, the reaction is as follows:

^GA +

The reaction was stopped for 3 days. The solvent was evaporated under reduced pressure and purified mjjjjlilililililililililililililili The product was subjected to analytical testing to obtain the following spectral data:

^{_{! H NMR (CDC1 3, 300}} MHz) δ 0.15 (s, 3H); 0.63-0.74 (m, 2H); 0.97 (d, J = 4.8 Hz, 3H); 1.00 (d, J = 5.1 Hz, 3H) ; 1.23 (t, J = 5.3 Hz, 6H); 1.67-1.84 (m, 8H); 2.03 (s, 3H), 2.37-2.48 (m, IH); 2.62-2.80 (m, 2H); 3.27 (s , 3H); 3.37 (s, 3H); 3.42-3.62 (m, 4H); 3,78 (q, J = 5.4 Hz, 4H); 4.28-4.34 (br, d, J = 4.3 Hz, IH); 4.39 (br, s, IH); 4.89 (s, br, IH); 5.19 (s, IH); 5.82-5.95 (m, 2H); 6.34-6.42 (m, IH); 6.59 (t, J = 8.7 Hz, IH); 6.96 (d, br, J = 8.7 Hz, IH); 7.28 (s, IH); 9.20 (s, 1H).

¹³ C NMR ( CDCI 3, 75 MHz ) δ -4.99; 11.19; 12.30; 12.60; 12.74; 18.39; 22.85; 23.52; 28.44; 32.25; 34.38; 35.01; 48.30; 56.69; 57.07; 58.25; 72.56; 81.19; 81.45; 108.24; 108.62; 126.56; 126.85; 132.68; 133.79; 134.96; 135.73; 141.43; 144.84; 156.06; 168.39; 180.49;

ESI (m/z): 742.3 (M + Na+).

It is proved that the product of the formula (5) is obtained:

OEt

^Si^ NH'GA formula ( $ ) Example 6

Geldanamycin (561 mg) plus 2-(diallylamino)ethylamine (200 mg), plus

The reaction was stopped for 2 days. The solvent was evaporated under reduced pressure and purified by silica gel column chromatography (eluent: EtOAc/EtOAc: EtOAc: EtOAc) The product was subjected to analytical testing to obtain the following spectral data:

^{_{! H NMR (CDC1 3, 300}} MHz) δ 0.93-1.05 (m, 6H); 1.66-1.92 (obscure, m, 6H); 2.03 (s, 3H); 2.35-2.47 (m, IH); 2.62-2.82 (m, 4H); 3.15 (d, J = 6.6 Hz, 4H); 3.27 (s, 3H); 3.37 (s, 3H); 3.40-3.45 (m, 2H); 3.46-3.68 (m, 2H); 4.32 (d, J = 9.9 Hz, IH); 4.53 (br, s, IH); 4.98 (s, br, 2H); 5.15-5.21 (m, 4H); 5.24 (s, IH); 5.78-5.96 ( m, 4H); 6.59 (t, J = 11.3 Hz, IH); 6.96 (d, J = 11.1 Ηζ, ΙΗ); 7.16 (s, br, IH); 7.27 (s, br, IH); 9.21 (s , 1H).

¹³ C NMR ( CDCI 3, 75 MHz ) δ 12.57; 12.62, 12.71, 22.95, 28.39, 32.28, 34.44, 35.12, 42.66, 50.75, 56.53, 56.70, 57.08, 72.54, 81.24, 81.52, 81.70, 108.40, 108.83, 118.22, 126.60, 126.81, 132.67, 133.88, 134.91, 135.03, 135.65, 141.27, 145.37, 156.03, 168.40, 180.13, 184.03.

ESI (m/z): 668.9 (M + H+).

It is proved that the product of the formula (6) is obtained:

Formula (6) Example 7

Geldanamycin (561 mg), plus 2-aminoindan (340 mg), plus dichloromethane (5 Add methanol (12 mL), add triethylamine (0.5 mL), the reaction formula is as follows:

The reaction was stirred at room temperature for 5.5 days, and the reaction was stopped. The solvent was evaporated under reduced pressure and purified by silica gel column chromatography (eluent: methylene chloride/methanol = 40:1). After re-dissolved in ethyl acetate and petroleum ether to give pure crystalline dropwise emphasis purple solid, named analysis tests for RZ816 (180 mg, 27%) 0 The product was the following spectral data:

^{_{! H NMR (GDC1 3, 300}} MHz) δ 0.92-1.06 (m, 6H); 1.57-1.67 (m, 3H); 1.74-1.86 (obsure, m, 4H); 2.03 (s, 3H); 2.33-2.46 (m, IH); 2.68-2.82 (m, 2H); 2.86-3.04 (m, 2H); 3.20-3.30 (m, 4H); 3.38 (s, 3H); 3.42-3.54 (m, 2H); -3.64 (m, IH); 4.26-4.38 (m, 2H); 4.85 (br, s, 2H); 5.20 (s, IH); 5.81-5.97 (m, 2H); 6.47-6.64 (m, 2H) ; 6.69 (d, J = 11.7 Hz, lH); 7.18-7.32 (m, 5H); 9.17 (s, 1H).

¹³ C NMR ( CDCI 3, 75 MHz ) δ 12.30; 12.57; 12.74; 22.86; 28.72; 32.24; 34.52; 34.93; 40.82; 41.32; 54.95; 56.67; 57.03; 72.56; 81.15; 81.36; 81.54; 108.61; 124.77; 126.50; 126.88; 127.15; 127.23; 132.77; 133.67; 134.90; 135.78; 139.56; 139.66; 141.18; 144.17; 156.15; 168.32; 180.66;

ESI (m/z): 684.4 (M + Na+).

Prove that the product shown in structure (7) is obtained:

Example 8

Geldanamycin (561 mg), adding 2-(2-naphthyloxy)ethylamine (700 m), adding dichloromethane (50 mL), stirring at room temperature, the reaction is as follows:

The reaction was stopped for 4 days. The solvent was evaporated under reduced pressure and purified (EtOAcjjjjjjjjjjjj The product was subjected to analytical testing to obtain the following spectral data:

¹ H NMR ( CDC1 ₃ , 300 MHz ) δ 0.94-1.08 (m, 6H); 1.66-1.88 (obscure, m, 6H); 2.03 (s, 3H); 2.35-2.52 (m, IH); 2.68-2.82 (m, 2H); 3.27 (s, 3H); 3.37 (s, 3H); 3.42-3.50 (m, IH); 3.54-3.64 (m, IH); 3.90-4.14 (m, 2H); 4.21-4.38 (m, 4H); 4.91 (s, br, 2H); 5.20 (s, IH); 5.81-5.95 (m, 2H); 6.59 (t, J = 11.6 Hz, IH); 6.62-6.71 (m, IH 6.95 (d, J = 11.1 Ηζ, ΙΗ); 7.14-7.22 (m, 2H); 7.29 (s, IH); 7.32-7.42 (m, IH); 7.41-7.50 (m, IH); 7.70- 7.82 (m, 3H); 9.13 (s, 1H).

¹³ C NMR ( CDCI 3, 75 MHz ) δ 12.35; 12.57; 12.76; 22.98; 28.56; 32.25; 34.36; 35.01; 45.01; 56.69; 57.05; 65.84; 72.53; 81.13; 81.56; 106.97; 108.91; 109.22; 118.53; 126.51; 126.56; 126.74; 126.88; 127.66; 129.26; 129.71; 132.79; 133.63; 134.25; 134.91; 135.77; 140.92; 144.94; 155.90; 156.12; 168.32; 180.76;

ESI (m/z): 738.4 (M + Na ⁺ ).

It is proved that the product of the formula (8) is obtained:

Example 9

Geldanamycin (561 mg), 2-(2,4-dichlorophenoxy)ethylamine (900 mg) was added, and dichloromethane (50 mL) was added. The reaction formula is as follows:

The color of the system is red. The reaction was stopped for 7 days. The solvent was evaporated under reduced pressure and purified (EtOAcjjjjjlilili The product was subjected to analytical testing to obtain the following spectral data:

^{_{! H NMR (CDC1 3, 300}} MHz) δ 0.94-1.06 (m, 6H); 1.66-1.86 (obscure, m, 6H); 2.30-2.44 (m, IH); 2.68-2.82 (m, 2H); 3.28 (s, 3H); 3.37 (s, 3H); 3.40-3.50 (m, IH); 3.52-3.64 (m, lH); 3.92-4.10 (m, 2H);

4.16- 4.26 (m, 3H); 4.32 (d, J = 9.6 Hz, IH); 4.86 (s, br, 2H); 5.20 (s, br, IH); 5.81-5.94 (m, 2H); 6.53- 6.64 (m, 2H); 6.85-7.00 (m, 2H);

7.17- 7.42 (m, 3H); 9.13 (s, 1H).

¹³ C NMR ( CDCI 3, 75 MHz ) δ 12.36; 12.56; 12.77; 22.96; 28.63; 32.26; 34.33; 35.00; 44.73; 56.69; 57.07; 67.89; 72.54; 81.11; 81.32; 81.56; 109.03; 109.68; 115.13; 126.51; 126.92; 127.16; 127.70; 130.26; 132.82; 133.58; 134.92; 135.81; 140.77; 145.02; 152.40; 156,10; 168.33; 180.85;

ESI (m/z): 756.4 (M + Na+).

Prove that the product shown in knot (9) is obtained:

Example 10

Geldanamycin (0.56 g ). Add 2-{[ ⁴ -(1, ² ,3-tris)butyl]phenoxy}ethylamine ( ^0.42 g), add dichloromethane (50 (mL), stir the reaction at room temperature. The reaction formula is as follows:

Stop the reaction. Evaporate the solvent under reduced pressure and purify by silica gel column chromatography Agent: Ethyl acetate/petroleum ether = 1 : 1 → dichloromethane / methanol = 30: 1 ) to give a purple solid, which was named RZ 820 (350 mg, 44%). The product was subjected to analytical testing to obtain the following spectral data:

^{! H NMR (CDC13, 300 MHz} ) δ 1.01 (d, J = 6.6 Hz, 6H); 1.55-1.68 (m, 2H); 1.70-1.84 (m, obscure, 4H); 1.86-2.00 (m, 4H) 2.03 (s, 3H); 2.34-2.47 (m, IH); 2.60 (t, J = 7.4 Hz, 2H); 2.66-2.80 (m, 2H); 3.27 (s, 3H); 3.37 (s, 3H) ); 3.42-3.50 (m, IH); 3.54-3.64 (m, IH); 3.80-4.05 (m, 2H); 4.10-4.21 (m, 2H); 4.25-4.35 (m, 2H); 4.40 (t , J = 7.1 Hz, 2H); 4.97 (s, br, 2H); 5.19 (s, IH); 5.81-5.94 (m, 2H); 6.53-6.66 (m, 2H); 6.84 (d, J = 8.4 Hz, 2H); 6.96 (d, J = 8.7 Hz, IH); 7.08 (d, J = 8.7 Hz, 2H); 7.27-7.32 (m, IH); 7.52 (s, IH); 7.70 (s, IH ); 9.16 (s, 1H).

¹³ C NMR ( CDC13, 75 MHz ) δ 12.39, 12.52, 12.74, 22.95, 28.24, 28.64, 29.65, 32.33, 34.19, 34.34, 35.10, 45.07, 49.94, 56.69, 57.07, 66.09, 72.63, 81.16, 81.43, 81.59, 108.91, 109.31, 114.75, 123.09, 126.46, 126.98, 129.42, 132.83, 113.62, 133.75, 134.65, 134.93, 135.86, 141.01, 145.09, 156.07, 156.38, 168.33, 180.84, 183,85.

ESI (m/z): 789.6 (M + H +); 811.5 (M + Na+).

Proof of the product:

Example 11

Geldanamycin (1 g), plus 1-tert-butoxy-reactive 3-(2-aminomethyl)piperidine (0.57 _g ), with dichloro:

The color of the system is orange-red. The reaction was carried out for 1.5 days and the color of the system was dark brown. Supplement 1 - Uncle Butoxycarbonyl-3-(2-aminomethyl)piperidine (0.20 g) was heated to reflux and allowed to react for 5 days to stop the reaction. The solvent was evaporated under reduced pressure and purified (EtOAcjjjjjjjjjjjj The product was a mixture of two epimers in a ratio of 1.5:1 (identified by HPLC). The product was subjected to analytical testing to obtain the following spectral data:

^X H NMR ( CDC13, 300 MHz ) δ 0.92-1.05 (m, 6H); 1.20-1.60 (m, obscure, 13H); 1.62-2.10 (m, obscure, 10H); 2.30-2.46 (m, IH); 2.62-2.80 (m, 2H); 2.81-2.95 (m, IH); 3.00-3.14 (m, IH); 3.27 (s, 3H); 3.37 (s, 3H); 3.40-3.52 (m, 2H); 3.54-3.63 (m, IH); 4.31 (d, J = 9.6 Hz, 2H); 5.05 (s, br, 2H); 5.18 (s, IH); 5.81-5.94 (m, 2H); 6.38 (s, Br, IH); 6.59 (t, J = 11.4 Hz, IH); 6.96 (d, J = 11.7 Hz, IH); 7.29 (s, IH); 9.17 (s, lH).

ESI (m/z): 765.7 (M +Na+).

Prove that the product shown in structure (11) is obtained:

Example 12

Geldanamycin (18) was added with 1-(2-decyloxy)-4-(2-aminoethyl)pyrazine (628 mg), and the mixture was stirred at room temperature. The reaction formula is as follows:

The reaction was stopped for 3 days. The solvent was evaporated under reduced pressure and purified (EtOAcjjjjjjjjjjjj The product was analyzed and tested to obtain the following wave transmission data:

^{_{! H NMR (CDC1 3, 300}} MHz) δ) 0.95-1.05 (m, 6H); 1.68-1.86 (m, 6H); 2.03 (s, 3H); 2.37-2.50 (m, IH); 2.65-2.82 ( m, 8H); 3.13 (br, s, 4H); 3.27 (s, 3H); 3.37 (s, 3H); 3.42-3.65 (m, 3H); 3.68-3.82 (m, IH); 3.87 (s, 3H); 4.32 (d, J = 10.2 Hz , IH); 4.50 (s, br, IH); 4.87 (s, br, 2H); 5.19 (s, IH); 5.81-5.96 (m, 2H); 6.59 (t, J = 11.3 Hz, IH); 6.85-7.06 (m, 5H); 7.19 (s, br, IH); 7.27 (s, IH); 9.21 (s, 1H).

¹³ C NMR ( CDC1 ₃ , 75 MHz ) δ 12.27; 12.57; 12.72; 23.07; 28.41 32.25; 34.41; 35.08; 41.75; 50.58; 52.66; 55.30; 55.81; 56.68; 57.04 72.50; 81.19; 81.44; 81.62; 108.67; 111.09; 118.29; 120.97 123.04; 126.56; 126.81; 132.68; 133.78; 134.97; 135.65; 141.02 141.15; 145.44; 152.16; 156.09; 168.38; 180.11;

ESI (m/z): 764.6 (M + H+); 786.6 (M + Na+).

Prove that the product is shown:

Formula (12)

Example 13

Geldanamycin (1 g), 2-[bis(2-decyloxy)amino]ethylamine (472 mg) was added, and dichloromethane (100 mL) was added, and the mixture was stirred at room temperature. The reaction formula is as follows:

The reaction was stopped for 5 days. The solvent was evaporated under reduced pressure and purified by chromatography (dield: EtOAc/EtOAc (EtOAc: EtOAc) The product was subjected to analytical testing to obtain the following spectral data:

lH NMR (CDCI3, 300 MHz) δ 0.92-1.04 (m, 6H); 1.62-1.84 (m, 6H); 2.03 (s, 3H); 2.36-2.50 (m, IH); 2.62-2.82 (m, 6H) 2.85-2.94 (m, 2H); 3.27 (s, 3H); 3.34 (s, 6H); 3.37 (s, 3H); 3.40-3.53 (m, 6H); 3.54-3.73 (m, 2H); 4.32 (d, J = 10.2 Ηζ, ΙΗ); 4.58 (s, br, IH); 4.83 (s br, 2H); 5.19 (s, IH); 5.80-5.98 (m , 2H); 6.59 (t, J = 11.3 Hz, IH) 6.96 (d, J = 11.4 Hz, lH); 7.20 (s, br, IH); 7.25 (s, IH); 9.22 (s, 1H).

¹³ C NMR ( CDC1 ₃ , 75 MHz ) δ 12.25; 12.59; 12.72; 22.93; 28.37 32.26; 34.35; 35.08; 43.09; 53.27; 53.68; 56.69; 57.06; 58.86; 71.45 72.50; 81.24; 81.52; 81.68; 108.50; 126.61; 126.76; 132.63 133.90; 135.02; 135.59; 141.21; 145.69; 156.08; 168.41; 179.83 184.11.

ESI (m/z): 705.4 (M + H+); 727.5 (M + Na+).

It is proved that the product of the formula (13) is obtained:

OMe

N, GA

MeO

Π式 (13)

Example 14

Geldanamycin (lg), [1-mercapto-2-(3,4-dimethoxy)phenyl]aminoethylamine (638 mg), dichloromethane (100 mL), room temperature Stir the reaction. The reaction formula is as follows:

The reaction was stopped for 2 days. The solvent was evaporated under reduced pressure and purified by silica gel column chromatography (eluent: EtOAc:EtOAc:EtOAc: Wave data:

^J H NMR ( CDCI 3, 300 MHz ) δ 0.96 (d, J = 6.0 Hz, 3H); 1.00 (d, J = 6.9 Hz, 3H); 1.72 (s, 3H); 1.75-1.88 (m, 4H); 2.03 (s, 3H); 2.33 (s, 3H); 2.35-2.44 (m, IH); 2.61-2.82 (m, 6H); 3.27 (s, 3H); 3.37 (s, 3H); 3.48-3.51 ( m, 2H); 3.53-3.56 (m, 2H); 3.85 (s, 3H); 3.87 (s, 3H); 4.32 (d, J = 9.9 Hz, 1H); 4.50 (s, br, 1H); 4.87 (s, br, 2H); 5.19 (s, 1H); 5.80-5.88 (m, 2H); 6.59 (t, J = 11.3 Hz, 1H); 6.64-6.81 (m, 3H); 6.97 (d, J = 11.4 Hz, 1H); 7.08 (s, br, 1H); 7.27 (s, 1H); 9.20 (s, 1H) .

¹³ C NMR ( CDC1 ₃ , 75 MHz ) δ 12.27; 12.57; 12.72; 22,94; 28.38; 32.25; 33.53; 34.35; 35.05; 41.32; 42.55; 55.28; 55.79; 55.86; 56.66; 57.04; 59.32; 72.51; 81.47; 81.62; 108.49; 108.80; 111.25; 111.88; 120.49; 126.56; 126.79; 132.51; 132.67; 133.80; 134.98; 135.64; 141.17; 145.35; 147.34; 148.81; 156.10; 168.39; 180.04;

ESI (m/z): 767.5 (M + H ⁺ ); 789.5 (M + Na+).

It is proved that the product of the formula (14) is obtained:

Formula (14)

Example 15 Detection of Biological Activity of Geld Derivatives

In order to carry out biological activity verification on each of the derivatives synthesized in the above Examples 1 - 14, the applicant used five kinds of cell forests to detect the growth inhibition rate of each derivative. Specifically, cell lines are used: Α549, MCF-7, AGS, HepG2 and A2780

Medium: HG-DMEM + 10% NBS medium (MCF-7 HCT116 and HepG2 cell lines); RPMI 1640 + 10% FBS medium (A2780, A549 and AGS cell lines)

Drug Dilution: All drugs were dissolved in DMSO and formulated into a 10 mg/ml stock solution, which was prepared by dilution with medium before administration.

ΙΟμΜ method for detecting growth inhibition rate

The cells were cultured on a flat-bottom 96-well plate; after 24 h, a final concentration of ΙΟμΜ was added. After 72 h, add 20 μl of MTT solution at a concentration of 5 mg/ml (dissolved in PBS, sterilized by filtration, stored at 4 ° C in the dark); incubate at 37 ° C for 4 h; aspirate MTT from 孑L with a lance Be careful not to touch the cells; add ΙΟΟμΙ 100% DMSO to each well; vortex for 15 min at room temperature; 570 nm wavelength reading (Thermo Multiskan Spectrum).

The results were calculated according to the formula (ODc-ODt) / ODc x 100%, where ODc is the control group OD ₅₇ . Value, ODt is the administration group OD ₅₇ . Values, specific inhibition results shown in Table 2. Table 2 _0: Growth geldanamycin derivative of 5 different cell lines inhibition assay results

In the table, A549, MCF-7, AGS, A2780 and HEPG-2 represent lung adenocarcinoma, breast cancer, gastric cancer, ovarian cancer and liver cancer cell lines, respectively, and represent the Geldardella shown in the chemical formulas (1) - (14). A derivative. The slightly larger black italic in the table is a positive result. The above results indicate that the compound of the present invention has excellent cell killing activity and cell growth inhibitory activity, and generally has excellent anticancer activity in vitro, especially for gastric cancer.

(AGS), ovarian cancer (A2780) and liver cancer (HEPG-2). Compared with the drug molecule RZC02 and RZC03, the compounds RZ823 and RZ824 have higher anticancer activity and broad-spectrum anticancer activity, which can inhibit multiple tumor cells at the same time. Although specific embodiments of the invention have been described in detail, those skilled in the art will understand. Various modifications and substitutions may be made to those details in light of the teachings of the invention, which are within the scope of the invention. The full scope of the invention is indicated by the appended claims and any equivalents thereof.

Claims

Rights request

A geldanamycin derivative selected from at least one of the compounds represented by the following formulas (1) to (14) or a salt thereof:

Formula (2)

-28-

Formula (8) /o//〇96lisosl>d sssonoiΜ

Εε --

O

U

O NH2

Formula (14)

2. A pharmaceutical composition, comprising at least the formula (1) - a compound represented by one of its salts or (I ⁴⁾ as an active ingredient.

3. The pharmaceutical composition of claim 2 further comprising a pharmaceutically acceptable excipient.

4. The pharmaceutical composition of claim 2 which further comprises other tumor therapeutic agents.

A cytostatic agent which contains at least one of the compounds represented by the chemical formulas (1) to (14) or a salt thereof as an active ingredient.

A cell proliferation inhibitor comprising at least one of a compound represented by the chemical formulas (1) to (14) or a salt thereof as an active ingredient.

7. Use of any of the geldanamycin derivatives or salts thereof of the formula (1) to (14) for the preparation of a medicament for treating a tumor.

8. Use of any of the geldanamycin derivatives or salts thereof of the formula (1) to (14) for the preparation of a killing agent for cell killing.

9. Use of any of the geldanamycin derivatives or salts thereof of the formula (1) to (14) for the preparation of a cell proliferation inhibitor.

A method for inhibiting cell growth, inhibiting cell proliferation or treating a tumor, which comprises administering to a subject a therapeutically effective amount of any geldanamycin derivative represented by the chemical formulas (1) to (14) or The steps of the salt.