CN1238338A - Dialkyl tin compound of mononuclear aromatic heterocycle hydroxamate and its synthesis - Google Patents

Abstract

A mononuclear aromatic heterocycle kind of dialkyl tin hydroxamate compound is synthesized through RCOOH + R"'OH--RCOOR"', RCOOR"'+NH2OH.HCl-KOH-RCONHOK-H+--RCONHOH, RCOOR"'+NH2OH.(1/2) H2SO4-NaOH--RCONHONa-H+-RCONHOH, R2 "SnO+2RCONR'OH--H2O+R2"Sn (OR'NOCR)2. It has low-poison, broad-spectrum and high anticancer activity when applied in low dosage (10 to the power -8 mol/L).

Description

Mononuclear aromatic heterocyclic monohydroyl hydroxamic acid dialkyl tin compound and synthesis thereof

The invention relates to an organotin compound with anticancer activity and a synthesis method thereof.

Gielen et al synthesized a series of mononuclear substituted dihydrocarbyltin benzoate compounds RR' Sn(OOCR”)₂(see the documents appl. organomet. chem.,1991,5, 497-506; 1993,7, 119-125; 1993,7, 201-206) generally has better in vitro anti-MCF-7 (breast cancer) and WiDr (colon cancer) activity than Cisplatin (Cisplatin), but the anti-cancer activity is limited because the compound is too toxic.

The aim of the invention is to develop a series of medicines at low doses (10)^-8mol/L) of the organic tin compound has broad spectrum, low toxicity and strong anticancer activity.

Synthesized mononuclear aromatic heterocyclic monohydroxy hydroxamic acid dialkyl tin compound R₂”Sn(OR’NOCR)₂20, up to hundreds being synthesized and ready to be synthesized. The structural general formula of the complex is confirmed by element analysis, infrared spectrum and nuclear magnetic resonance hydrogen spectrum:wherein, R "= Et, Bu, Ph, etc.; r' = H, Ph, etc.;

and the like, and benzene rings substituted at various positions with halogen (F, Cl, Br, I), nitro, amino, hydroxyl, alkyl, alkoxy, and the like. (some of the compounds are shown in Table 1) 1. synthetic route (the synthetic route in Table 1 is as follows:) [3]2. Preparation method (Each route of the synthetic routes was performed as follows)<1>]a：

0.2 mol of the acid RCOOH is dissolved in 2 mol of methanol CH₃OH or 1.334 mol of absolute ethanol C₂H₅In OH, cooling in an ice-water bath, slowly dripping 16.7 ml of concentrated sulfuric acid while stirring, and refluxing for 4 hours. Cooled to 30 ℃, adjusted to PH =9 with 20% sodium carbonate solution, extracted with anhydrous ether, the combined extracts, the ether extracts dried over anhydrous magnesium sulfate, shaken, and left to stand overnight. Filtering to remove desiccant, evaporating the filtrate with rotary evaporator to remove diethyl ether and excessive alcohol, and collecting the rest fraction by vacuum distillation to obtain ester RCOOR. Route of road[1]b：

0.1 mol of the acid RCOOH is dissolved in 0.8 mol of absolute ethanol C₂H₅In OH, cooling in ice water bath, slowly dripping 7 ml of concentrated sulfuric acid while stirring, and refluxing for 6 hours. Cooled to 30 ℃, adjusted to PH =9 with 20% sodium hydroxide solution, extracted with toluene, the extracts were combined, the toluene extract was dried over anhydrous magnesium sulfate, shaken and allowed to stand overnight. Filtering to remove desiccant, evaporating the filtrate with rotary evaporator to remove toluene and excessive alcohol, and collecting the rest by vacuum distillation to obtain ester RCOOR (RCOOC)₂H₅)。

TABLE 1 synthetic route assignments for the class of compounds synthesized

Note: the abbreviated ligands in the table correspond to the names as follows:

BHA = Benzohydroxamic acid, benzoyl hydroxamic acid;

SHA = Salicylhydroxamic acid;

PHBHA = p-Hydroxybenzohydroxamic acid, p-hydroxybenzoyl hydroxamic acid;

CiHA = cinnamyl hydroxamic acid;

N-PhBHA = N-Phenyl benzoic hydroxamic acid, N-phenylbenzoyl hydroxamic acid;

ONBHA = o-nitrobenzoic hydroxamic acid, o-nitrobenzoyl hydroxamic acid;

MNBHA = m-nitrobenzoylhydroxamic acid, m-nitrobenzoyl hydroxamic acid;

PNBHA = p-nitrobenzoic hydroxamic acid, p-nitrobenzoyl hydroxamic acid;

OABHA = o-Aminobenzohydroxamic acid, anthranoylhydroxamic acid;

mabba = m-Aminobenzohydroxamic acid, m-aminobenzoyl hydroxamic acid;

PABHA = p-Aminobenzohydroxamic acid, p-aminobenzoyl hydroxamic acid;

FuHA = Furan-2-carbohydroxamic acid, furoyl hydroxamic acid;

NiHA = Nicotinohydroxamic acid, nicotinoyl hydroxamic acid;

IsHA = Isonicotinohydroxamic acid, isonicotinoyl hydroxamic acid. Route [2]a:

24.32 g (0.35 mol) of hydroxylamine hydrochloride were dissolved in 140 ml of methanol by reflux, and 30.86 g (0.55 mol) of potassium hydroxide were dissolved in 100 ml of methanol by the same method. And when the solution and the solution are cooled to 30-40 ℃, pouring the rear solution into the front solution while cooling and shaking, standing for 5 minutes to ensure that the potassium chloride is completely precipitated, and filtering out the precipitate to obtain a methanol solution of hydroxylamine and potassium hydroxide. Adding 0.2 mol of the prepared ester RCOOR into the filtrate, stirring for 2 hours to obtain RCONHOK precipitate, filtering, dissolving in a small amount of water, stirring, acidifying with 2N acetic acid to pH =5, stirring for 2 hours, filtering to obtain a crude product, recrystallizing with water twice, and vacuum drying to obtain ligand hydroxamic acid RCONHOH. Route [2]b:

19.6 g (0.49 mol) of sodium hydroxide was dissolved in 80 ml of water, and slowly added to 100 ml of ice water containing 15.8 g (0.1 mol) of hydroxylamine sulfate, and 0.1 mol of ester RCOOR. RTM. was added. Stir at room temperature under nitrogen for 4 hours overnight. Acidification with 25% sulfuric acid in an ice bath resulted in precipitation. Filtering, recrystallizing twice with water, and vacuum drying to obtain ligand hydroxamic acid RCONHOH. Route [3]:

8 mmol of ligand hydroxamic acid RCONR' OH are dissolved by reflux in a mixture of 150 ml of toluene and 50 ml of absolute ethanol (or 200 ml of 3: l benzene: methanol) and 4 mmol of tin dihydrocarbyloxide R are added₂"SnO, reflux for six hours. After the reaction is finished, evaporating a half of the solvent, evaporating the rest under the evacuation state to obtain a crude product, recrystallizing by using absolute ethyl alcohol, filtering, collecting, and drying in vacuum to obtain the mononuclear aromatic heterocyclic monohydroxyhydroxamic acid dialkyl tin compound R₂”Sn(OR’NOCR)₂(when R = substituent-OH, -NH₂The benzene ring of (a), the whole reaction process needs to be protected by nitrogen).

The elemental analysis and physical property data of the ligand RCONR' OH and the complex are shown in Table 2, the infrared spectrum data are shown in Table 3, and the nuclear magnetic hydrogen spectrum data are shown in Table 4. 3. The apparatus used for the experiment: digital melting point instrument produced by Shanghai physical optical instrument factory(ii) a 240C elemental analyzer and Vario EL type elemental analyzer; shimadzu IR-435 infrared spectrometer and Perkin-Elmer-983 infrared spectrometer;¹h NMR is measured by a Bruker DRX300MHZ nuclear magnetic resonance instrument and a Bruker AM-500MHZ nuclear magnetic resonance instrument, TMS is used as an internal standard, and deuterated DMSO and deuterated chloroform areused as solvents; the molecular weight is determined by freezing point depression method, and the solvent is naphthalene.

TABLE 2 elemental analysis and physical Property data for ligands and complexes

Compound (I)	Molecular weight	Appearance of the product	mp/℃	Yield of	Elemental analysis% found value (calculated value)
								Mr	％	C	H	N
	BHA			137	White colour	128.7～129.7	65						61.52(61.31)	5.13(5.11)	10.38(10.22)
[(BHA)2SnEt2]		448	White colour					254.3～254.4	85	48.08(48.14)	4.72(4.94)	6.17(6.24)
	[(BHA)2SnBu2]			505	White colour	225.1～226.7	75						51.96(52.26)	5.89(5.94)	5.78(5.54)
[(BHA)2SnPh2]		545	White colour					123.9～125.8	83	57.70(57.23)	3.91(4.04)	5.46(5.14)
	SHA			153	White colour	177.3～179.0	60						54.75(54.90)	4.69(4.58)	8.97(9.15)
[(SHA)2SnEt2]		481	White colour					256.0～258.4	81	45.36(44.94)	4.69(4.61)	5.87(5.82)
	[(SHA)2SnBu2]			537	Light yellow	210.0 decomposition	70						49.71(49.15)	5.88(5.58)	5.49(5.21)
[(SHA)2SnPh2]		577	White colour					Decomposition > 300	88	54.57(54.06)	3.47(3.81)	4.90(4.85)
	PHBHA			153	White colour	179.2～180.8	67						55.21(54.90)	4.56(4.58)	8.93(9.15)
[(PHBHA)2SnEt2]		481	White colour					195.0 decomposition	78	45.77(44.94)	4.53(4.61)	5.29(5.82)
	[(PHBHA)2SnBu2]			537	White colour	245.6～246.4	81						48.64(49.15)	5.76(5.58)	5.10(5.21)
[(PHBHA)2SnPh2]		577	White colour					Decomposition > 300	84	53.57(54.06)	3.84(3.81)	4.97(4.85)
	CiHA			163	Shallow powder	97.4～99.3	80						66.07(66.26)	5.39(5.52)	8.70(8.59)
[(CiHA)2SnBu2]		557	Shallow powder					138.8～140.3	83	56.11(56.01)	6.18(6.10)	5.16(5.03)
	[(N-PhBHA)2SnBu2]			657	Pink colour	94.6～96.4	89						62.21(62.10)	5.70(5.78)	4.21(4.26)
ONBHA		182	White colour					148.0～149.3	25	46.81(46.15)	3.27(3.30)	14.78(15.38)
	[(ONBHA)2SnBu2]			595	Light yellow	230.1～231.6	62						43.98(44.36)	4.97(4.70)	9.13(9.40)
MNBHA		182	White colour					153.4～154.9	60	46.49(46.15)	3.27(3.30)	15.47(15.38)
	[(MNBHA)2SnBu2]-			595	Light yellow	252.3～253.8	74						43.89(44.36)	4.96(4.70)	9.08(9.40)
PNBHA		182	Light yellow					166.0～167.8	50	46.44(46.15)	3.29(3.30)	14.81(15.38)

[(PNBHA)2SnBu2]	595	Yellow colour	240.0～241.7	73	43.97(44.36)	4.89(4.70)	9.17(9.40)
								OABHA	152	Shallow powder	146.7～148.0	54	55.08(55.26)	5.21(5.26)	18.32(18.42)
[(OABHA)2SnBu2]	535	White colour	＞300	76	49.25(49.35)	5.83(5.98)	10.60(10.47)
								MABHA	152	White colour	153.4～154.8	60	55.38(55.26)	5.09(5.26)	18.38(18.42)
[(MABHA)2SnBu2]	535	White colour	＞300	71	49.27(49.35)	5.82(5.98)	10.40(10.47)
								PABHA	152	Shallow powder	169.9～170.3	85	55.23(55.26)	5.16(5.26)	18.30(18.42)
[(PABHA)2SnBu2]	535	Shallow powder	166.8～167.4	83	49.27(49.35)	5.80(5.98)	10.32(10.47)
								FuHA	127	White colour	119.9～121.6	63	47.14(47.24)	3.83(3.94)	10.89(11.02)
[(FuHA)2SnBu2]	485	White colour	174 break down	83	44.63(44.54)	5.26(5.36)	5.87(5.77)
								NiHA	138	White colour	161.4～161.9	61	52.35(52.17)	4.25(4.34)	20.18(20.29)
[(NiHA)2SnBu2]	507	White colour	189.0～191.0	74	47.38(47.34)	5.68(5.52)	10.92(11.05)
								IsHA	138	White colour	160.6～160.9	65	52.30(52.17)	4.36(4.34)	20.32(20.29)
[(IsHA)2SnBu2]	507	Rice yellow	142.2～143.7	84	47.27(47.34)	5.72(5.52)	10.86(11.05)

TABLE 3 Infrared Spectroscopy data (vcm) of ligands and complexes^-1)

Compound (I)	vNH-OH	vC=O	vN-O	vSn-C	vSn-O
						BHA	3060～2900	1650	895	----	----
[(BHA)2SnEt2]	3397	1590 1567	950 927	548	470
						[(BHA)2SnBu2]	3427	1604 1569	916	550	497
[(BHA)2SnPh2]	3415	1600	913	538	450
						SHA	3380～2700	1640	910	----	----
[(SHA)2SnEt2]	3421	1599 1577	950 921	540	459
						[(SHA)2SnBu2]	3420	1597	962 916	520	475
[(SHA)2SnPh2]	3395	1598	950 916	539	450
						PHBHA	3270～2675	1650	908	----	----
[(PHBHA)2SnEt2]	3301	1600 1569	960 916	537	430
						[(PHBHA)2SnBu2]	3298	1603	963	551	469
[(PHBHA)2SnPh2]	3309	1600	949 916	561	449
						CiHA	3230～2600	1650	980	----	----
[(CiHA)2SnBu2]	----	1572	1030	563	471
						N-PhBHA	3150～2900	1620	908	----	----
[(N-PhBHA)2SnBu2]	----	1540	920	532	492
						ONBHA	3220～3010	1650	890	----	----
[(ONBHA)2SnBu2]	3400	1590	950	----	----
						MNBHA	3350～3200	1640	890	----	----
[(MNBHA)2SnBu2]	3400	1580	920	----	----
						PNBHA	3250～2800	1650	890	----	----
[(PNBHA)2SnBu2]	----	1560	920	----	----
						OABHA	3250～2800	1650	890	----	----
[(OABHA)2SnBu2]	----	1580	910	543	471
						MABHA	3250～2780	1645	890	----	----
[(MABHA)2SnBu2]	----	1580	910	541	458
						PABHA	3250～2790	1640	890	----	----
[(PABHA)2SnBu2]	----	1600	908	573	468
						FuHA	3300～2500	1630	935	----	----
[(FuHA)2SnBu2]	----	1590	945	593	469
						NiHA	3160 2800～2500	1640	900	----	----
[(NiHA)2SnBu2]	----	1530	910	503	472
						IsHA	3200～2500	1635	900	----	----
[(IsHA)2SnBu2]	----	1530	918	601	494

4. Structural characterization (1) Infrared Spectroscopy

As can be seen from table 3: the infrared spectrum of the ligand RCONR' OH is 3380-2500 cm^-1The range of the absorption peak of the hydroxyl stretching vibration is 1620-1650 cm^-1A carbonyl absorption peak appeared. When the ligand forms a complex, the hydroxyl absorption peak disappears, which shows that the deprotonation of the hydroxyl group is coordinated with the oxygen atom thereof, and the carbonyl absorption peak is red-shifted to 1530-1604cm^-1The carbonyl oxygen is illustrated as forming a coordinate bond with metallic tin. In the complex v_N-HShifting to high frequency 3400cm^-1The coordination of nitrogen atoms in NHOH groups and tin is eliminated, and the tin belongs to hard acid, so that the soft and hard acid-base rules are met. V of partial complex_N-HDisappearance, probably because active hydrogen on a nitrogen atom in the partial ligand is transferred to an oxygen atom and exists in a form of HO-C = N-OH, and the oxygen atom is coordinated with tin after dehydrogenation, so 3300cm is not observed in the complex^-1Left and right v_N-HPeak(s). The coordinated N-O stretching vibration absorption peak generally moves to high frequency by delta v 10-55 cm^-1And the absorption strength is increased, which on the one hand excludes coordination of the nitrogen atom in the NH-OH group and on the other hand also proves that chlorine in the NH-OH coordinates with tin. Only v is observed in the IR spectrum of the complex_Sn-OAbsorption peak, no vobserved indeed_Sn-NAbsorption peak, about 415cm^-1To (3).

The above IR parameters indicate that the ligand is chelated to tin with the oxygen atom in the CO-NHOH (or HO-C = N-OH) group. Only one v is observed in the complex_Sn-CIndicating that the two hydrocarbon radicals R' are in the trans position.

TABLE 4 nuclear magnetic hydrogen spectra of ligands and complexesData (delta ppm)

Compound (I)	Ring (C)	CH3	CH2	(CH2)nSn	-NHOH(-NHO-)
						BHA	7.49～7.89(5H)	----	----	----	10.21(2H)
[(BHA)2SnEt2]	7.48～7.67(10H)	1.15(6H)	----	1.35(4H)	9.83(2H)
						[(BHA)2SnBu2]	7.38～7.75(10H)	0.83(6H)	1.35(4H)	1.46(8H)	9.72(2H)
[(BHA)2SnPh2]	7.31～7.82(10H)	7.25(10H)Ph			9.85(2H)
						SHA	6.99～7.31(4H)	----	----	----	unobserved
[(SHA)2SnEt2]	6.82～8.17(8H)	1.10(6H)	----	1.34(4H)	8.72(2H)
						[(SHA)2SnBu2]	6.78～7.59(8H)	0.84(6H)	1.32(4H)	1.46(8H)	8.81(2H)
[(SHA)2SnPh2]	6.80～8.12(8H)	7.37(10H)Ph			8.70(2H)
						PHBHA	6.97～7.82(4H)	----	----	----	9.53(1H)8.70(1H)
[(PHBHA)2SnEt2]	6.62～7.43(8H)	1.14(6H)	----	1.37(4H)	8.41(2H)
						[(PHBHA)2SnBu2]	6.75～7.58(8H)	0.81(6H)	1.30(4H)	1.48(8H)	8.45(2H)
[(PHBHA)2SnPh2]	6.84～7.81(8H)	7.32(10H)Ph			8.66(2H)
						CiHA	6.47～7.56(7H) Ph-CH=CH-	----	----	----	unobserved
[(CiHA)2SnBu2]	6.48～7.54(14H)	0.87(6H)	1.29(8H)	1.51(4H)	----
						N-PhBHA	7.16～7.63(10H)	----	----	----	10.72(1H)
[(N-PhBHA)2SnBu2]	7.25～7.29(20H)	0.94(6H)	1.46(4H)	1.78(8H)	----
						ONBHA	7.90～8.67(4H)	----	----	----	11.67(1H)9.37(1H)
[(ONBHA)2SnBu2]	7.68～7.88(8H)	1.01(6H)	1.44(4H)	1.72(8H)	----
						MNBHA	7.91～8.71(4H)	----	----	----	11.76(1H)9.51(1H)
[(MNBHA)2SnBu2]	7.80～8.73(8H)	1.00(6H)	1.45(4H)	1.73(8H)	----
						PNBHA	8.14～8.48(4H)	----	----	----	11.74(1H)9.51(1H)
[(PNBHA)2SnBu2]	8.10～8.32(8H)	0.99(6H)	1.45(4H)	1.72(8H)	----
						OABHA	6.47～7.28(4H)	----	----	----	10.16(1H)8.80(1H)
[(OABHA)2SnBu2]	6.51～7.32(8H)	0.91(6H)	1.39(4H)	1.56(8H)	----
						MABHA	6.64～7.04(4H)	----	----	----	10.94(1H)8.85(1H)
[(MABHA)2SnBu2]	6.67～7.10(8H)	0.91(6H)	1.38(4H)	1.57(8H)	----
						PABHA	6.53～7.45(4H)	----	----	----	10.71(1H)8.64(1H)
[(PABHA)2SnBu2]	6.66～7.56(8H)	0.90(6H)	1.35(4H)	1.58(8H)	----
						FuHA	6.55～7.75(3H)	----	----	----	11.10(1H)9.06(1H)
[(FuHA)2SnBu2]	6.71～7.54(6H)	0.90(6H)	1.25(4H)	1.64(8H)	----
						NiHA	7.50～8.90(4H)	----	----	----	11.39(1H)9.22(1H)
[(NiHA)2SnBu2]	7.45～9.16(8H)	0.91(6H)	1.25(4H)	1.69(8H)	----
						IsHA	7.65～8.69(4H)	----	----	----	11.50(1H)9.30(1H)
[(IsHA)2SnBu2]	7.73～8.79(8H)	0.90(6H)	1.25(4H)	1.70(8H)	----

(2): nuclear magnetic hydrogen spectrum analysis

As can be seen from table 4: the absorption peak of benzene ring or heterocyclic ring proton in all complexes is obviously changed compared with that of free ligand, which is caused by the induction effect after the ligand is coordinated with tin.

The ligand has two protons respectively in the range of 9.53-11.76 and 8.64-9.53 ppm before coordination, which are respectively assigned to OH and NH of NH-OH, but in the complex, the OH proton peak of NH-OH has disappeared, which proves that the coordination of deprotonation of hydroxyl group in NH-OH and oxygen atom is consistent with the presumption of IR spectrum. After BHA, SHA and PHBHA are coordinated, NH peak of NH-OH still exists, on one hand, coordination of nitrogen atoms in NH-OH groups is eliminated, and on the other hand, coordination of oxygen in NH-OH and tin is also proved. Some preparationsNo absorption signal appears in the proton on the nitrogen of the compound, which is probably related to the rapid exchange of active hydrogen, the active hydrogen on the nitrogen atom in the ligand is transferred to an oxygen atom and exists in the form of HO-C = N-OH, and the oxygen atom is coordinated with tin after dehydrogenation, so that no NH proton peak is observed in the complex. Therefore, the temperature of the molten metal is controlled,¹h NMR results also demonstrate the presumption that the ligand is coordinated to the tin chelate by the oxygen atom in the CO-NHOH (or HO-C = N-OH) group. In addition, hydrocarbyl R 'appears as a multiplet, also indicating that R' is in a non-linear inversion.

In combination with the above-mentioned IR,¹the compound is considered to be in a hexa-coordinated mononuclear distorted octahedral structure by parameters such as H NMR and element analysis. The anticancer activity of the compound (shown as the general formula)

The compounds are subjected to activity screening by the Beijing medical university natural medicine and bionic medicine national key laboratories (tables 5,6,7A,7B,8A and 8B) and the Shanghai pharmaceutical institute New drug screening national key laboratories (tables 9 and 10) of Chinese academy of sciences, and are found to have potent activity.

TABLE 5 median inhibitory concentration IC of the complexes on four human tumor cells₅₀(umol/L)

Compound (I)	Human nasopharyngeal carcinoma KB	Human leukemia HL-60	Human colon cancer HCT-8	Ehrlich ascites Ehrlich ascite
					[(BHA)2SnEt2]	4.21	4.20	3.05	1.21
[(BHA)2SnBu2]	1.30	1.32	0.73	0.24
					[(BHA)2SnPh2]	3.98	Is inactive	1.10	Is free ofActivity of
[(SHA)2SnEt2]	Is inactive	0.57	1.81	0.39
					[(SHA)2SnBu2]	1.52	0.31	0.25	0.11
[(SHA)2SnPh2]	Is inactive	Is inactive	Is inactive	Is inactive
					[(PHBHA)2SnEt2]	3.95	0.54	1.47	0.22
[(PHBHA)2SnBu2]	1.17	0.21	0.31	0.12
					[(PHBHA)2SnPh2]	Is inactive	Is inactive	2.63	Is inactive

TABLE 6 different concentrationsThe inhibition rate of the compound on the growth of tumor cells%

Compound (I)	P388	A549	SGC
				1.0μM 10μM 100μM	1.0μM 10μM 100μM	1.0μM 10μM 100μM
	[(BHA)2SnEt2]	74.0 98.1 100	57.1 90.8 91.8				0 70 88.9
[(BHA)2SnBu2]				91.3 100 100	64.4 93.9 92.9	12.2 90 94.4
	[(BHA)2SnPh2]	100 100 100	73.5 86.7 87.7				12.5 94.4 97.8
[(SHA)2SnEt2]				58.9 100 100	58.7 93.9 94.4	2.2 92.2 95.6
	[(SHA)2SnBu2]	100 100 100	54.1 92.9 93.9				3.3 93.3 95.6
[(PHBHA)2SnEt2]				100 100 100	61.2 94.4 93.9	7.8 87.8 91.1
	[(PHBHA)2SnBu2]	100 100 100	86.7 93.9 95.7				50 80.0 91.1

Tables 7A-7B, tables 8A-8B evaluation of results: -ineffective + less effective + + significant + + + more effective

TABLE 7 inhibition of tumor cell growth by different concentrations of the complexes%

Compound (I)	Testing Serial number	K562 *	Bel-7402*
				0.1. mu.M 1. mu.M 10. mu.M evaluation	0.1. mu.M 1. mu.M 10. mu.M evaluation
		[(BHA)2SnEt2]	11			40.03 59.70 81.02 ++	2.56 6.69 82.87 +
[(BHA)2SnBu2]	6			42.82 74.66 80.98 ++	14.38 59.30 88.19 ++
		[(BHA)2SnPh2]	3			-8.52 7.17 81.04 +	1.49 42.78 92.92 +
[(SNA)2SnEt2]	9			27.85 36.44 72.88 +	2.42 12.98 72.83 +
		[(SHA)2SnBu2]	4			33.25 65.88 80.18 ++	21.47 66.85 93.23 ++
[(SHA)2SnPh2]	1			-35.21 -6.31 9.57 -	-1.77 -2.70 2.92 -
		[(PHBHA)2SnEt2]	10			5.86 32.09 77.74 +	8.48 23.17 82.57 +
[(PHBHA)2SnBu2]	5			23.01 57.17 75.52 ++	20.11 59.78 94.26 ++
		[(PHBHA)2SnPh2]	2			2.14 3.25 48.22 -	2.57 7.95 74.96 +

Note that: human erythroleukemia K₅₆₂Adopting a tetrazolium salt (MTT) colorimetric method;

human liver cancer Bel-7402 adopts a Sulfarhodamine (SRB) protein staining method.

TABLE 7 inhibition of tumor cell growth by different concentrations of the complexes%

Testing Serial number	BGC*	KB*	HCT-8*
				0.1. mu.M 1. mu.M 10. mu.M evaluation	0.1. mu.M 1. mu.M 10. mu.M evaluation	0.1. mu.M 1. mu.M 10. mu.M evaluation
	11	-15.40 -23.46 52.13 +	-1.32 3.40 68.43 +				6.93 7.58 98.56 +
6				-28.32 22.88 82.79 +	4.35 41.21 88.09 +	16.24 52.57 95.85 ++
	3	-6.97 7.84 91.84 +	0.00 0.95 76.37 +				12.97 48.78 97.86 +
9				-19.67 -12.56 17.77 -	10.59 -2.08 38.75 -	Not determined
	4	-22.44 2.61 95.21 +	6.62 24.20 97.54 +				29.08 66.44 97.33 ++
1				-9.95 -23.70 -24.41 -	3.41 0.20 23.09 -	3.85 3.19 -0.79 -
	10	-26.30 -30.81 45.26 -	-4.16 2.84 74.29 +				Not determined
5				- 10.85 94.08 +	Not determined	Not determined
	2	-0.47 -1.66 3.55 -	-6.05 -5.67 11.91 -				6.62 15.91 91.40 +

Note that^*: human gastric cancer BGC (MTT method); human nasopharyngeal carcinoma KB (MTT method); human colon cancer HCT-8(SRB method)

TABLE 8 inhibition of tumor cell growth by different concentrations of the complexes%

TransformingCompound (I)	Testing Serial number	KB*	BGC-823
				0.1. mu.M 1. mu.M 10. mu.M evaluation	0.1. mu.M 1. mu.M 10. mu.M evaluation
		[(CiHA)2SnBu2]
[(N-PhBHA)2SnBu2]	98203710			40.99 91.10 98.26 ++	-10.19 49.32 94.48 +
		[(OABHA)2SnBu2]	990071			Not determined	18.68 95.30 98.79 ++
[(MABHA)2SnBu2]	990073			2.03 -1.83 -4.57 -	7.93 -9.58 -1.20 -
		[(PABHA)2SnBu2]	98203670			45.18 83.62 92.33 ++	12.28 44.38 82.34 +
[(FuHA)2SnBu2]	98203610			37.81 85.60 96.65 ++	15.56 77.18 97.48 ++
		[(NiHA)2SnBu2]	98203630			46.73 85.81 98.18 ++	21.56 72.78 98.18 ++
[(IsHA)2SnBu2]	98203650			36.37 90.46 98.03 ++	19.67 65.24 98.46 ++

Note that^*: human nasopharyngeal carcinoma KB (SRB method); human gastric cancer BGC-823(SRB method).

TABLE 8 inhibition of tumor cell growth by different concentrations of the complexes%

Testing Serial number	Bel-7402*	HCT-8*	HL-60*
				0.1. mu.M 1. mu.M 10. mu.M evaluation	0.1. mu.M 1. mu.M 10. mu.M evaluation	0.1. mu.M 1. mu.M 10. mu.M evaluation
98203710				-18.24 21.00 94.78 +	8.61 54.14 83.09 ++	-28.16 55.97 79.42 ++
	990071	24.75 95.99 98.87 ++	10.71 83.13 92.91 ++				10.34 93.72 95.32 ++
990073				1.32 2.84 1.77 -	0.46 0.55 2.87 -	-15.02 -4.31 8.86 -
	98203670	21.69 37.48 81.86 +	17.38 52.87 92.42 ++				3.90 61.37 76.66 ++
98203610				36.15 70.07 95.10 ++	10.18 64.98 89.51 ++	25.90 64.12 84.88 ++
	98203630	41.60 56.50 94.77 ++	11.07 58.28 89.66 ++				35.78 65.38 84.20 ++
98203650				40.48 55.06 94.32 ++	14.03 64.91 92.98 ++	1.55 52.49 71.68 ++

Note that^*: human liver cancer Bel-7402(SRB method); human colon cancer HCT-8(SRB method); human leukemia HL-60 (M)TT method).The screening method comprises the following steps: tetrazolium salt (MTT) reduction cell line: p₃₈₈Action time of mouse leukemia: and (6 h) evaluating the result: and (4) invalidation: 10^-5mol/L＜85％；

Weak effect: 10^-5mol/L is more than or equal to 85 percent or 10^-6mol/L＞50％；

The strong effect is as follows: 10^-6mol/L is more than or equal to 85 percent or 10^-7mol/L＞50％；

TABLE 9 inhibition of tumor cell growth by different concentrations (mol/L) of the complex%

Compound (I)	Sample numbering	10-4	10-5	10-6	10-7	10-8	Evaluation of
								[(CiHA)2SnBu2]	1107	94.6	96.8	97.8	97.8	97.8	High effect
[(N-PhBHA)2SnBu2]	1090	85.0	96.3	77.5	67.5	62.5	High effect
								[(ONBHA)2SnBu2]	1033	71.4	52.2	50.0	47.6	48.8	Weak effect
[(MNBHA)2SnBu2]	1035	84.5	54.4	55.6	52.2	52.2	High effect
								[(PNBHA)2SnBu2]	1037	78.9	60.0	57.8	53.3	53.3	High effect
[(OABHA)2SnBu2]	1098	92.7	98.2	98.2	98.2	96.4	High effect
								[(MABHA)2SnBu2]	1100	84.6	88.5	88.5	88.5	86.5	High effect
[(PABHA)2SnBu2]	1086	93.8	97.5	66.3	66.3	45.0	High effect
								[(FuHA)2SnBu2]	1080	85.9	88.7	93.0	91.5	91.5	High effect
[(NiHA)2SnBu2]	1082	87.3	90.1	90.1	93.0	91.5	High effect
								[(IsHA)2SnBu2]	1084	87.3	90.1	90.1	90.5	91.5	High effect

The screening method comprises the following steps: sulfbrhodamine B (SRB) protein staining method cell strain: a-549 human lung adenocarcinoma action time: and (5) evaluating the result: and (4) invalidation: 10^-5mol/L＜85％；

Weak effect: 10^-5mol/L is more than or equal to 85 percent or 10^-6mol/L＞50％；

The strong effect is as follows: 10^-6mol/L is more than or equal to 85 percent or 10^-7mol/L＞S0％；

TABLE 10 inhibition of tumor cell growth by different concentrations (mol/L) of the complex%

Compound (I)	Sample numbering	10-4	10-5	10-6	10-7	10-8	Evaluation of
								[(CiHA)2SnBu2]	1107	100.0	97.8	98.9	63.7	13.2	High effect
[(N-PhBHA)2SnBu2]	1090	94.8	92.7	93.8	92.7	89.6	High effect
								[(ONBHA)2SnBu2]	1033	90.8	89.7	85.5	85.5	85.5	High effect
[(MNBHA)2SnBu2]	1035	90.8	92.0	85.5	85.5	83.9	High effect
								[(PNBHA)2SnBu2]	1037	94.3	90.8	90.8	89.7	83.9	High effect
[(OABHA)2SnBu2]	1098	100.0	98.8	97.5	100.0	92.6	High effect
								[(MABHA)2SnBu2]	1100	95.3	95.3	97.6	74.0	9.4	High effect
[(PABHA)2SnBu2]	1086	94.8	91.7	91.7	86.5	14.6	High effect
								[(FuHA)2SnBu2]	1080	92.9	87.9	90.9	89.9	87.9	High effect
[(NiHA)2SnBu2]	1082	92.9	90.9	91.9	91.9	25.3	High effect
								[(IsHA)2SnBu2]	1084	91.9	90.9	90.9	89.9	79.8	High effect

The screening method comprises the following steps: sulfbrhodamine B (SRB) protein staining method, tetrazolium salt (MTT) reduction method cell strain: HO-8910 human ovarian cancer, SPC-A4 human lung cancer, HCT-116 human colon cancer action time: and (5) evaluating the result: and (4) invalidation: 10^-5mol/L＜85％；

Weak effect: 10^-5mol/L is more than or equal to 85 percent or 10^-6mol/L＞50％；

The strong effect is as follows: 10^-6mol/L is more than or equal to 85 percent or 10^-7mol/L＞50％；

TABLE 11 inhibition of tumor cell growth by cisplatin at various concentrations (mol/L)%

Cell line	10-4	10-5	10-6	10-7	10-8	Evaluation of
							HO-8910 SPC-A4 HCT-116	84.4 48.7 78.8	56.3 8.0 0.0	0.4 0.4 0.0	4.7 0.0 0.0	0.0 0.0 0.0	Invalidation Invalidation Invalidation

As can be seen from the test results in tables 5 to 11, the organotin compounds of the present invention have a broad spectrum, low toxicity and potent anticancer activity: (1) broad spectrum: the series of compounds can be used for treating human nasopharyngeal carcinoma KB, human gastric cancer BGC-823, and human liverCancer Bel-7402, human colon cancer HCT-8, human leukemia HL-60, human erythroleukemia K₅₆₂Ehrlich ascites, P₃₈₈Mouse leukemia, A-549 human lung adenocarcinoma tumor cells and the like have stronger inhibitory power. (2) Low toxicity: taking Wish-human amniotic cells as an example, the SRB method is used, the action time is 72 hours, and the toxic concentration of the series of compounds to normal cells is 10^-6mol/L, inhibition concentration of 10 to tumor cells^-8The difference in mol/L is two orders of magnitude. (3) The strong effect is as follows: the series of compounds P₃₈₈The inhibition rate of mouse leukemia and A-549 human lung adenocarcinoma tumor cells is strong, but the inhibition rate of the cisplatin which is widely applied to clinic at present to HO-8910 human ovarian cancer, SPC-A4 human lung cancer and HCT-116 human colon cancer is ineffective (see table 11).

Example 1:

complex [ (FuHA)]₂SnBu₂]The structure is as shown in the foregoing, whereinR' = H, R "= Bu. (1): synthetic route

[3](2): preparation Process route [1]a: (Synthesis of furoic acid methyl ester)

22.4 g (0.2 mol) of furoic acid are dissolved in 64 g (2 mol, 80 ml) of methanol CH₃In OH, cooling in an ice-water bath, slowly dripping 16.7 ml of concentrated sulfuric acid while stirring, and refluxing for 4 hours. Cooled to 30 ℃, adjusted to PH =9 with 20% sodium carbonate solution, extracted with anhydrous ether, the combined extracts, the ether extracts dried over anhydrous magnesium sulfate, shaken, and left to stand overnight. And filtering to remove a drying agent, quickly evaporating diethyl ether and excessive alcohol from orange filtrate by using a rotary evaporator, and collecting 14mL of orange fraction at 86-86.5 ℃/30mmHg by using reduced pressure distillation on the rest to obtain the methyl furoate with the yield of 62.3 percent. Route [2]a: (synthetic FuHA)

24.32 g (0.35 mol) of hydroxylamine hydrochloride were dissolved in 140 ml of methanol by reflux, and 30.86 g (0.55 mol) of potassium hydroxide were dissolved in 100 ml of methanol by the same method. To be treatedCooling the two solutions to 30-40 deg.C, shaking while cooling, adding the latter solution into the former solution, standing for 5 min to ensure the potassium chloride is completely precipitated, and filtering to remove precipitate to obtain methanol solution of hydroxylamine and potassium hydroxide. To this filtrate was added 24.66 g of the prepared methyl furoate (0.2 mol, 21 ml), stirred for 2 hours to give a white precipitate of furoyl hydroxamate potassium salt, filtered off, dissolved in a small amount of water, stirred, acidified with 2N acetic acid to PH =5, stirred for 2 hours, filtered to give a crude product, recrystallized twice with water, and vacuum dried to give white ligand furoyl hydroxamate FuHA. Route [3]: (Synthesis of Complex [ (FuHA)]₂SnBu₂])

1.0168 g (8 mmol) of FuHA ligand furoyl hydroxamate were dissolved in a mixture of 150 ml of toluene and 50 ml of absolute ethanol under reflux, and 0.996 g (4 mmol) of Bu dibutyl tin oxide was added₂SnO, refluxing for 6 hours. After the reaction is finished, evaporating half of the solvent, evaporating the rest under the vacuum condition to obtain a crude product, recrystallizing with anhydrous ethanol, filtering, collecting, and vacuum drying to obtain a white powdery complex [ (FuHA)]₂Snu₂]. (3): use of

The compound has strong inhibitory effect on human nasopharyngeal carcinoma KB, human gastric cancer BGC-823, human liver cancer Bel-7402, human colon cancer HCT-8, and human leukemia HL-60, and can be used for treating P₃₈₈The inhibition rate of mouse leukemia and A-549 human lung adenocarcinoma tumor cells is shown inHigh effect. Example 2:

complex [ (OABHA)₂SnBu₂]The structure is as shown in the specification, wherein,

r' = H, R "= Bu (1): synthetic route

[3](2): preparation Process route [1]b: (Synthesis of Ethyl anthranilate)

13.71 g (0.1 mol) of anthranilic acid is dissolved in 36.8 g (0.8 mol, 46 ml) of absolute ethyl alcohol, cooled in an ice-water bath, slowly dropped into 7 ml of concentrated sulfuric acid under stirring, and heated and refluxed for 6 hours. Cooled to 30 ℃, adjusted to PH =9 with 20% sodium hydroxide solution, extracted with toluene, the extracts were combined, the toluene extract was dried over anhydrous magnesium sulfate, shaken andallowed to stand overnight. Filtering to remove a drying agent, quickly evaporating toluene and excessive alcohol from colorless filtrate by using a rotary evaporator, and collecting 8 ml of colorless fraction at the temperature of 116-117 ℃/5mmHg by using reduced pressure distillation on the rest to obtain ethyl anthranilate with the yield of 53.2%. Route [2]b: (Synthesis OABHA)

19.6 g (0.49 mol) of sodium hydroxide was dissolved in 80 ml of water, and slowly added to 100 ml of ice water containing 15.8 g (0.1 mol) of hydroxylamine sulfate, and 16.52 g (0.1 mol) of ethyl anthranilate was added. Stir at room temperature under nitrogen for 4 hours overnight. Acidification with 25% sulfuric acid in an ice bath resulted in precipitation. Filtering, recrystallizing twice with water, and vacuum drying to obtain light pink powder OABHA. Route [3]: (Synthesis [ (OABHA)₂SnBu₂])

1.2172 g (8 mmol) of ligand anthranilic hydroxamic acid OABHA were dissolved in a mixture of 150 ml of toluene and 50 ml of absolute ethanol (or 200 ml of a mixture of 3: 1 benzene: methanol) under reflux, followed by 0.996 g (4 mmol) of dibutyltin oxide Bu₂SnO is refluxed for 6 hours under the protection of nitrogen. After the reaction is finished, evaporating half of the solvent, evaporating the rest under the condition of vacuum pumping to obtain crude product, recrystallizing with anhydrous ethanol, filtering, collecting, and vacuum drying to obtain white complex [ (OABHA)₂SnBu₂]. (3): use of

The compound has strong inhibitory effect on human nasopharyngeal carcinoma KB, human gastric cancer BGC-823, human liver cancer Bel-7402, human colon cancer HCT-8, and human leukemia HL-60, and can be used for treating P₃₈₈The inhibition rate of mouse leukemia and A-549 human lung adenocarcinoma tumor cells is strong.

Claims (2)

1. A mononuclear aromatic heterocyclic monohydroyl hydroxamic acid dialkyl tin compound is characterized in that the structural general formula is as follows:r "= Et, Bu, Ph; r 'is'H, Ph; r is a heterocyclic ring

And

and benzene rings substituted by nitro, amino, hydroxyl at different positions.

2. The method for synthesizing a tin compound according to claim 1, characterized by comprising the steps of:

0.2 mol of the acid RCOOH is dissolved in 2 mol of methanol CH₃OH or 1.334 mol of absolute ethanol C₂H₅In OH, cooling in an ice-water bath, slowly dripping 16.7 ml of concentrated sulfuric acid while stirring, and refluxing for 4 hours. Cooled to 30 ℃, adjusted to PH =9 with 20% sodium carbonate solution, extracted with anhydrous ether, the combined extracts, the ether extracts dried over anhydrous magnesium sulfate, shaken, and left to stand overnight. Filtering to remove desiccant, quickly evaporating diethyl ether and excessive alcohol from the filtrate with rotary evaporator, and collecting the rest fraction by vacuum distillation to obtain ester RCOOR; or

0.1 mol of the acid RCOOH is dissolved in 0.8 mol of absolute ethanol C₂H₅In OH, cooling in ice water bath, slowly dripping 7 ml of concentrated sulfuric acid while stirring, and refluxing for 6 hours. Cooled to 30 ℃, adjusted to PH =9 with 20% sodium hydroxide solution, extracted with toluene, the extracts were combined, the toluene extract was dried over anhydrous magnesium sulfate, shaken and allowed to stand overnight. Filtering to remove desiccant, evaporating the filtrate with rotary evaporator to remove toluene and excessive alcohol, and collecting the rest by vacuum distillation to obtain ester RCOOR (RCOOC)₂H₅)；

24.32 g (0.35 mol) of hydroxylamine hydrochloride were dissolved in 140 ml of methanol by reflux, and 30.86 g (0.55 mol) of potassium hydroxide were dissolved in 100 ml of methanol by the same method. And when the solution and the solution are cooled to 30-40 ℃, pouring the rear solution into the front solution while cooling and shaking, standing for 5 minutes to ensure that the potassium chloride is completely precipitated, and filtering out the precipitate to obtain a methanol solution of hydroxylamine and potassium hydroxide. Adding 0.2 mol of the obtained ester RCOOR into the filtrate, stirring for 2 hr to obtain RCONHOK precipitate, filtering, dissolving in small amount of waterWater, stirring, acidifying with 2N acetic acid to pH =5, stirring for 2 hours, filtering to obtain a crude product, recrystallizing with water twice, and vacuum drying to obtain ligand hydroxamic acid RCONHOH; or

19.6 g (0.49 mol) of sodium hydroxide was dissolved in 80 ml of water, and slowly added to 100 ml of ice water containing 15.8 g (0.1 mol) of hydroxylamine sulfate, and 0.1 mol of ester RCOOR. RTM. was added. Stir at room temperature under nitrogen for 4 hours overnight. Acidification with 25% sulfuric acid in an ice bath resulted in precipitation. Filtering, recrystallizing with water twice, and vacuum drying to obtain ligand hydroxamic acid RCONHOH; (3)

8 mmol of ligand hydroxamic acid RCONR' OH are dissolved in a mixture of 150 ml of toluene and 50 ml of absolute ethanol (or 200 ml of a mixture of 3: 1 benzene: methanol) by reflux, and 4 mmol of tin dihydrocarbyloxide R are added₂"SnO, reflux for six hours. After the reaction is finished, evaporating a half of the solvent, evaporating the rest under the evacuation state to obtain a crude product, recrystallizing by using absolute ethyl alcohol, filtering, collecting, and drying in vacuum to obtain the mononuclear aromatic heterocyclic monohydroxyhydroxamic acid dialkyl tin compound R₂”Sn(OR’NOCR)₂(when R = substituent-OH, -NH₂The benzene ring of (a), the whole reaction process needs to be protected by nitrogen).