Cyclohexene compounds and use thereof

Abstract

A cyclohexene compound and use thereof, which relates to the technical field of pharmaceutical chemistry is provided. The cyclohexene compound is the cyclohexene compound having a structural formula I or a pharmaceutically acceptable salt, an ester group prodrug or a stereoisomer thereof. It is verified by activity assay for influenza NA that the cyclohexene compound has a great inhibiting activity against both wild-typed and drug-resistant influenza viruses, and the inhibiting activity IC.sub.50 of some compounds against wild-typed and drug-resistant influenza viruses is lower than 5 nM, showing significant inhibiting effect on influenza viruses. This shows that the cyclohexene compound or a pharmaceutically acceptable salt or a stereoisomer thereof has superior application prospect on preparing anti-influenza drugs, thereby providing a new drug choice for treating influenza clinically.

Claims

1. A cyclohexene compound having a structural formula II or a pharmaceutically acceptable salt, a stereoisomer thereof: ##STR00086## wherein, m is selected from 0 or 1; R is selected from H; R.sub.1 is selected from amino, or guanidyl; R.sub.2 is selected from C.sub.1-C.sub.4 alkyl; R.sub.3 and R.sub.4 are independently selected from H; R.sub.5 is absent, or R.sub.5 is selected from H, C.sub.1-C.sub.4 alkyl, substituted C.sub.1-C.sub.4 alkyl, C.sub.3-C.sub.6 cycloalkyl, substituted C.sub.3-C.sub.6 cycloalkyl, C.sub.3-C.sub.6 heterocyclyl, or substituted C.sub.3-C.sub.6 heterocyclyl; n is selected from 2; X is selected from O, NHCOO, or OCONR.sub.9R.sub.10; and R.sub.9 and R.sub.10 are independently selected from H, C.sub.1-C.sub.4 alkyl, substituted C.sub.1-C.sub.4 alkyl, C.sub.3-C.sub.6 cycloalkyl, or substituted C.sub.3-C.sub.6 cycloalkyl, or R.sub.9 and R.sub.10 together with the atom to which they are attached form a C.sub.3-C.sub.6 heterocyclic ring containing O and NH.

2. The cyclohexene compound or a pharmaceutically acceptable salt, or a stereoisomer thereof according to claim 1, wherein the cyclohexene compound is a compound having formula III: ##STR00087##

3. The cyclohexene compound or a pharmaceutically acceptable salt, or a stereoisomer thereof according to claim 1, wherein, the substituted C.sub.1-C.sub.4 alkyl is selected from hydroxyethyl, methoxyethyl, ethoxyethyl, or cyclopropylmethyl; and the substituted C.sub.3-C.sub.6 heterocyclyl is selected from morpholine-4-carbonyloxy4.

4. The cyclohexene compound or a pharmaceutically acceptable salt, or a stereoisomer thereof according to claim 1, wherein the cyclohexene compound is a compound selected from one of the following compounds: ##STR00088## ##STR00089## ##STR00090##

5. A pharmaceutical composition, comprising the cyclohexene compound or a pharmaceutically acceptable salt, or a stereoisomer thereof according to claim 1, and a pharmaceutically acceptable excipient or carrier.

6. The cyclohexene compound or a pharmaceutically acceptable salt, or a stereoisomer thereof according to claim 2, wherein, the substituted C.sub.1-C.sub.4 alkyl is selected from hydroxyethyl, methoxyethyl, ethoxyethyl, or cyclopropylmethyl; and the substituted C.sub.3-C.sub.6 heterocyclyl is selected from morpholine-4-carbonyloxy.

Description

DETAILED DESCRIPTION

(1) The term alkyl herein refers to saturated hydrocarbyl or unsaturated chained alkyl, chained alkyl refers to linear or branched alkyl. For example, C.sub.1C.sub.4 alkyl refers to saturated or unsaturated, linear or branched alkyl having 14 carbon atoms, wherein examples of saturated linear alkyl comprise but are not limited to ethyl, n-propyl and so on; examples of saturated branched alkyl comprise but are not limited to isopropyl group, tertiary butyl and so on; examples of unsaturated linear alkyl comprise but are not limited to ethenyl, propenyl and so on, and examples of unsaturated branched alkyl comprise but are not limited to 2-methylpropenyl and so on; cycloalkyl refers to alkyl having cyclic structure, for example, C.sub.3C.sub.6 cycloalkyl refers to saturated or unsaturated cyclic alkyl having 36 carbon atoms, wherein examples of saturated cycloalkyl comprise but are not limited to cyclopropyl, cyclobutyl, cyclopropyl substituted with methyl, and examples of unsaturated cycloalkyl comprise but are not limited to cyclopentene and so on. The term heterocyclic group refers to a group which is monocyclic, bicyclic or tricyclic, wherein one or more carbon atoms on the ring are independently and optionally substituted by heteroatoms selected from N, O, P, S and so on, such as, pyrrolidyl, tetrahydrofuryl, dihydrofuryl, tetrahydrothienyl, tetrahydropyranyl, dihydropyranyl, tetrahydrothiapyranyl, piperidyl, morpholinyl, thiomorpholinyl, thioxane group, thiazolidine group, oxazolidine group,or piperazinyl.

(2) The term substituted means that hydrogen group in a specific structure is replaced by a specified substituent.

(3) The present disclosure comprises the compound of formula I-IV in a free form and also comprises a pharmaceutically acceptable salt and stereoisomer thereof. The pharmaceutically acceptable salt of the present disclosure can be synthesized from the compounds comprising alkaline or acidic parts of the present disclosure through common chemical methods. In general, the salt of alkaline compound is synthesized through ion exchange chromatography or through a reaction between free alkaline and inorganic or organic acid in a form of desirable salt, which are in chemical calculated amount or excess amount in suitable solvents or combinations thereof. Similarly, the salt of acidic compound is prepared through a reaction with a suitable inorganic or organic alkaline.

(4) Therefore, the pharmacologically acceptable salt of the present disclosure includes conventional non-toxic salt of the present disclosure synthesized by the reaction between the alkaline compounds of the present disclosure and inorganic or organic acids. For example, the conventional non-toxic salt includes the salt prepared from inorganic acids, such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, or nitric acid, and also includes the salt prepared from organic acids, such as acetic acid, propionic acid, succinic acid, glycolic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, maleic acid, hydroxy maleic acid, benzene acetic acid, glutamic acid, benzoic acid, salicylic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid, hydroxyethanesulphonic acid, or trifluoroacetic acid.

(5) If the compound of the present disclosure is acidic, the appropriate pharmaceutically acceptable salt means a salt prepared from a pharmaceutically acceptable non-toxic alkaline comprising inorganic or organic alkaline. The salt prepared from inorganic alkaline includes ammonium salt, calcium salt, lithium salt, magnesium salt, potassium salt, sodium salt, zinc salt, etc., and ammonium salt, calcium salt, magnesium salt, potassium salt and sodium salt. The salt prepared from organic non-toxic alkaline includes primary amine, secondary amine, tertiary amine, (quaternary ammonium) salt, and substituted amines comprising natural substituted amines, cyclic amines and alkaline ion exchange resin such as arginine, choline, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethyl piperidine, glucosamine, histidine, isopropylamine, lysine, methylglucosamine, morpholine, piperazine, piperidine polyamine resin, triethylamine, trimethylamine, tripropylamine, or tromethamine.

(6) In addition to the conventional methods disclosed in the literatures or exemplified in the experimental procedures, the compound of the present disclosure can be prepared by the method described in the following synthetic approaches. The compound and synthetic method described in the present disclosure can be better understood in combination with the following synthesis schemes. The synthesis schemes describe methods for preparing compounds of the present disclosure, and the methods are merely illustrative descriptions for purpose of explanation without limitation of the scope of the disclosure.

(7) The method for preparing a starting material ethyl (3R, 4R, 5S)-4-acetamido-5-azido-3-acetoxy-1-cyclohexene-1-carboxylate in the following examples is seen from J. Am. Chem. Soc., 1997(119): 681-690.

EXAMPLE 1

(8) a) Preparation of ethyl (3R,4S,5S)-4-acetamido-5-azido-3-((3S)-3-(2-methoxylethoxyl)piperidin)-1-cyclohexene-1-carboxylate.

(9) ##STR00005##

(10) According to the above equation, 20 mmol of ethyl (3R,4R,5S)-4-acetamido-5-azido-3-acetoxyl-1-cyclohexene-1-carboxylate and 1 mmol of tetrakis (triphenylphosphine) palladium were added into a dry twin-neck flask, and after the air in the system was replaced with nitrogen gas twice, 40 mL of redistilled DMF (dimethylformamide) was added with an injector and well stirred. 40 mmol of DIPEA (N,N-Diisopropylethylamine) was then added and stirred, after it is cooled to 0 C., the solution (40 mL) of (3S)-3-(2-methoxylethoxyl)piperidin trifluoroacetate in DMF was slowly dripped into. The mixture was stirred at 0 C. for 20 minutes after the dripping, then it was placed in an oil bath to react at 70 C. for 1 hour. Gradually cooled it to 0 C. once the TLC (DCM:MeOH=10:1) indicated the reaction was completed. Water in the same volume was slowly dripped into, and after the mixture was stirred well, it was extracted with EA (ethyl acetate) and evaporated to dryness. DMF is removed by using an oil pump, and a brown viscous object was obtained. After an elution through column purification (MeOH: DCM=1:50), 3.8 g yellow-white powder solid was obtained, with a yield of 46.4%.

(11) The characterization analysis of the product:

(12) .sup.1HNMR(400 MHz, CDCl.sub.3):ppm6.880(s,1H,NH),5.736-5.717(dd,1H,2-CH),4.221-4.187(q,2H,CH.sub.2CH.sub.3),3.895-3.868(dd,1H,4-CH),3.855-3.780(dd,1H,3-CH),3.754-3.487(t,4H, OCH.sub.2CH.sub.2O),3. 381(s,3H.sub.2OCH.sub.3),2.907-2.851(m,1H,NCH.sub.2CHOCH.sub.2),2.815-2.790(d,1H,NCH.sub.2CHOCH.sub.2),2.731-2.705(t,1H,NCH.sub.2CH.sub.2),2.519-2.496(t,1H,NCH.sub.2CH.sub.2),2.293-2.271(d,1H,NCH.sub.2CHOCH.sub.2),2.244-2.207(dd,1H,5-CH),2.037-1.956(dd,1H,6-CH.sub.2),1.933(s,3H,COCH.sub.3),1.797-1.714(dd,1H,6-CH.sub.2),1.704-1.685(m,1H,NCH.sub.2CH.sub.2CH.sub.2),1.393-1.368(m,2H,NCH.sub.2CH.sub.2CH.sub.2),1.315-1.300(m,1H,NCH.sub.2CH.sub.2CH.sub.2),1.298-1.280(t,3H,CH.sub.2CH.sub.3).

(13) ESI-MS m/z: 410.3(M+H).sup.+.

(14) b) Preparation of ethyl (3R,4R,5S)-4-acetamido-5-amino-3-((3S)-3-(2-methoxylethoxyl)piperidin)-1-cyclohexene-1-carboxylate.

(15) ##STR00006##

(16) According to the above equation, ethyl (3R,4S,5S)-4-acetamido-5-azido-3-((3S)-3-(2-methoxylethoxyl)piperidin) -1-cyclohexene-1-carboxylate was added into a single-neck flask, and Raney Ni in catalytic amount and 40 mL of absolute ethyl alcohol were then added and well stirred. The air in the system was replaced with hydrogen gas, the mixture was continually stirred at room temperature for 2 hours. After the TLC (DCM:MeOH=10:1) indicated the reaction was completed, Raney Ni was filtered out with diatomite. The solvent was then evaporated, purified by column (DCM:MeOH=5:1), and a white foam solid was obtained.

(17) The characterization analysis of the product:

(18) .sup.1H-NMR(400 MHz,CDCl.sub.3):ppm6.883(s,1H,NH),5.574-5.513(dd,1H,2-CH), 4.197-4.171(q,2H,CH.sub.2CH.sub.3),3.839-3.816(dd,1H,4-CH),3.793-3.790(dd,1H,3-CH),3.644-3.591(t,4H,OCH.sub.2CH.sub.2O),3.555-3.508(dd,1H,5-CH),3.369(s,3H, OCH.sub.3),3.251-3.227(m,1H,NCH.sub.2CHOCH2),2.896-2.890(d,1H,NCH.sub.2CHOCH.sub.2),2.884-2.871(t,1H,NCH.sub.2CH.sub.2),2.858-2.850(t,1H,NCH.sub.2CH.sub.2),2.837-2.796(d,1H,NCH.sub.2CHOCH.sub.2),2.700-2.673(dd,1H,6-CH.sub.2),2.515-2.468(dd,1H,6-CH.sub.2),2.174-2.123(m,3H,NCH.sub.2CH.sub.2CH.sub.2, NCH.sub.2CH.sub.2CH.sub.2), 1.988(s,3H,COCH.sub.3), 1.792-1.690(m,1H,NCH.sub.2CH.sub.2CH.sub.2), 1.314-1.295(t,3H,CH.sub.2CH.sub.3).

(19) ESI-MS m/z: 384.2(M+H).sup.+.

(20) c) Preparation of ethyl (3R,4R,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3S)-3-(2-methoxylethoxyl)piperidin)-1-cyclohexene-1-carboxylate.

(21) ##STR00007##

(22) According to the above equation, 0.5 mmol of ethyl (3R,4R,5S)-4-acetamido-5-amino-3-((3S)-3-(2-methoxylethoxyl)piperidin)-1-cyclohexene-1-carboxylate was added into a single-neck flask, and 2 mL of DCM and 0.75 mmol of TEA were then added and stirred at room temperature till completely dissolved. 0.75 mmol of Boc.sub.2O was dripped into and stirred at room temperature for 30 minutes. After the TLC (DCM:MeOH=20:1) indicated the reaction was completed, small amount of water was added for extraction with DCM, and anhydrous sodium sulfate was used for drying before the solvent being evaporated to dryness. After being purified by column (DCM:MeOH=20:1), a white solid was obtained. ESI-MS m/z for the product: 484.2(M+H).sup.+.

(23) d) Preparation of (3R,4R,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3S)-3-(2-methoxyl ethoxyl)piperidin)-1-cyclohexene-1-carboxylic acid.

(24) ##STR00008##

(25) According to the above equation, 160 mg of ethyl (3R,4R,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3S)-3-(2-methoxylethoxyl)piperidin)-1-cyclohexene-1-carboxylate was added into a single-neck flask, and 5.5 ml of 1,4-dioxane, 0.5 ml of water and 0.5 ml of 1N KOH aqueous solution were then added and stirred overnight at room temperature. After the TLC (DCM:MeOH=20:1) indicated that the reaction was completed, the solvent was evaporated to dryness, and dried by using an oil pump. Methanol was used for dissolving, and pH was regulated to 5 with acid resin. After being filtered, evaporated to dryness, and purified by the column (DCM:MeOH=5:1), a white solid was obtained.

(26) The characterization analysis of the product:

(27) .sup.1H-NMR(400 MHz,MeOD):ppm6.777(dd,1H,2-CH),4.058-4.007(dd,1H,4-CH), 3.684-3.671(dd,1H,5-CH),3.660-6.652(dd,1H,3-CH),3.628-3.620(t,4H.sub.2OCH.sub.2CH.sub.2O),3.360(s,3H,OCH.sub.3),2.976-2.951(m,1H,NCH.sub.2CHOCH.sub.2),2.926-2.902(d,1H,NCH.sub.2CHOCH.sub.2),2.763-2.708(t,1H,NCH.sub.2CH.sub.2),2.622-2.610(t,1H,NCH.sub.2CH.sub.2),4.452-2.490(d,1H,NCH.sub.2CHOCH.sub.2),2.246-2.176(dd,1H,6-CH.sub.2),1.970(s,3H,COCH.sub.3),1.950-1.900(dd, 1H,6-CH.sub.2),1.790-1.775(m,1H,NCH.sub.2CH.sub.2CH.sub.2),1.595-1.525(m,1H,NCH.sub.2CH.sub.2CH.sub.2), 1.442(s,9H,O(CH.sub.3).sub.3),1.361-1.292(m,2H, NCH.sub.2CH.sub.2CH.sub.2,NCH.sub.2CH.sub.2CH.sub.2).

(28) ESI-MS m/z: 456.2(M+H).sup.+.

(29) e) Preparation of (3R,4R,5S)-4-acetamido-5-amino-3-((3S)-3-(2-methoxylethoxyl)piperidin)-1-cyclohexene-1-carboxylic acid trifluoroacetate.

(30) ##STR00009##

(31) According to the above equation, (3R,4R,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3S)-3-(2-methoxylethoxyl)piperidin)-1-cyclohexene-1-carboxylic acid was added into a single-neck flask, and 4 mL of DCM and 1 mL of TFA were then added and stirred at room temperature for 1 hour. After the TLC (DCM:MeOH=5:1) indicated that the reaction was completed, the solvent was evaporated to dryness and the residual TFA was removed by using an oil pump. Diethyl ether was added, and filtered out when a solid appears, and a white powder is obtained.

(32) The characterization analysis of the product:

(33) .sup.1H-NMR(400 MHz,D.sub.2O):ppm6.938(dd,1H,2-CH),4.696-4.644(dd,1H,4-CH), 4.457-4.434(dd,1H,3-CH),3.989-3.980(dd,1H,5-CH),3.628-3.620(t,2H,OCH.sub.2CH.sub.2O),3.634-3.630(m,1H,NCH.sub.2CHOCH.sub.2),3.588-3.572(t,2H, OCH.sub.2CH.sub.2O),3.554-3.536(m,4H,NCH.sub.2CHOCH.sub.2,NCH.sub.2CH.sub.2),3.396(s,3H,OCH.sub.3), 3.112-3.068(dd,2H,6-CH.sub.2),2.637-2.266(m,1H, NCH.sub.2CH.sub.2CH.sub.2),2.200-2.168(m,2H,NCH.sub.2CH.sub.2CH.sub.2,NCH.sub.2CH.sub.2CH.sub.2),2.139(s,3H,COCH.sub.3),1.865-1.831(m,1H,NCH.sub.2CH.sub.2CH.sub.2).

(34) ESI-MS m/z: 356.2(M+H).sup.+.

EXAMPLE 2

(35) a) Preparation of ethyl (3R,4S,5S)-4-acetamido-5-azido-3-((3R)-3-(2-methoxylethoxyl)piperidin)-1-cyclohexene-1-carboxylate.

(36) ##STR00010##

(37) Ethyl (3R,4S,5S)-4-acetamido-5-azido-3-((3R)-3-(2-methoxylethoxyl)piperidin)-1-cyclohexene-1-carboxylate was prepared with reference to the Example 1a).

(38) The characterization analysis of the product:

(39) .sup.1-H-NMR(400 MHz,CDCl.sub.3):ppm6.837(s,1H,NH),5.715-5.694(dd,1H,2-CH), 4.201-4.184(q,2H,CH.sub.2CH.sub.3),3.925-3.877(dd,1H,4-CH),3.781-3.715(dd,1H,3-CH),3.604-3.530(t,2H,OCH.sub.2CH.sub.2O), 3.502-3.491(m,1H,NCH.sub.2CHOCH.sub.2), 3.490-3.451(t,2H,OCH.sub.2CH.sub.2O), 3.351 (s,3H,OCH.sub.3),3.282-3.239(d,1H,NCH.sub.2CHOCH.sub.2),3.016-2.991(t,1H,NCH.sub.2CH.sub.2),2.893-2. 837(t,1H,NCH.sub.2CH.sub.2),2.529-2.516(m,2H,NCH.sub.2CHOCH.sub.2,5-CH),2.352-2.285(dd,1H,6-CH.sub.2),2.248-2.202(m,1H,NCH.sub.2CH.sub.2CH.sub.2),2.182-2.177(dd,1H,6-CH.sub.2),2.061(s,3H,COCH.sub.3),1.959-1.935(m,1H,NCH.sub.2CH.sub.2CH.sub.2),1.715-1.760(m,1H,NCH.sub.2CH.sub.2CH.sub.2),1.532-1.423(m,1H,NCH.sub.2CH.sub.2CH.sub.2),1.352-1.330(t,3H,CH.sub.2CH.sub.3).

(40) ESI-MS m/z: 410.3 (M+H).sup.+.

(41) b) Preparation of ethyl (3R,4R,5S)-4-acetamido-5-amino-3-((3R)-3-(2-methoxylethoxyl)piperidin)-1-cyclohexene-1-carboxylate.

(42) ##STR00011##

(43) Ethyl (3R,4R,5S)-4-acetamido-5-amino-3-((3R)-3-(2-methoxylethoxyl)piperidin)-1-cyclohexene-1-carboxylate was prepared with reference to Example 1b).

(44) The characterization analysis of the product:

(45) .sup.1-H-NMR(400 MHz,CDCl.sub.3):ppm6.837(s,1H,NH),5.555-5.534(dd,1H,2-CH), 4.206-4.126(q,2H,CH.sub.2CH.sub.3),3.892-3.818(dd,1H,4-CH),3.655-3.559(t,2H,OCH.sub.2CH.sub.2O),3.520-3.495(t,2H,OCH.sub.2CH.sub.2O),3.452-2.430(dd,1H,3-CH),3.395-3.356(s,3H,OCH.sub.3),3.295-3.250(dd,1H,5-CH),3.249-3.210(m,1H,NCH.sub.2CHOCH.sub.2),3.021-2.999(d,1H,NCH.sub.2CHOCH.sub.2),2.925-2.883(t,1H,NCH.sub.2CH.sub.2),2.870-2.857(t,1H,NCH.sub.2CH.sub.2),2.615-2.570(d,1H,NCH.sub.2CHOCH.sub.2),2.544-2.517(dd,1H,6-CH.sub.2),2.165-2.155(dd,1H,6-CH.sub.2),2.142-2.118(m,1H,NCH.sub.2CH.sub.2CH.sub.2),2.044(s,3H,COCH.sub.3),1.949-1.920(m,1H,NCH.sub.2CH.sub.2CH.sub.2),1.714-1.704(m,1H,NCH.sub.2CH.sub.2CH.sub.2),1.525-1.400(m,1H,NCH.sub.2CH.sub.2CH.sub.2),1.314-1.195(t,3H,CH.sub.2CH.sub.3).

(46) ESI-MS m/z: 384.2(M+H).sup.+.

(47) c) Preparation of ethyl (3R,4R,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3R)-3-(2-methoxylethoxyl)piperidin)-1-cyclohexene-1-carboxylate.

(48) ##STR00012##

(49) Ethyl (3R,4R,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3R)-3-(2-methoxylethoxyl)piperidin)-1-cyclohexene-1-carboxylate was prepared with reference to Example 1 c). ESI-MS m/z for the product: 484.2(M+H).sup.+.

(50) d) Preparation of (3R,4R,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3R)-3-(2-methoxylethoxyl)piperidin)-1-cyclohexene-1-carboxylic acid.

(51) ##STR00013##

(52) (3R,4R,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3R)-3-(2-methoxylethoxyl)piperidin)-1-cyclohexene-1-carboxylic acid was prepared with reference to Example 1 d).

(53) The characterization analysis of the product:

(54) .sup.1H-NMR(400 MHz,MeOD):ppm6.751(dd,1H,2-CHH),5.491(s,1H,NH)4.070-4.019(dd,1H,4-CH),3.693-3.680(dd,1H,5-CH),3.632-3.621(t,2H,OCH.sub.2CH.sub.2O),3.526-3.516(t,2H,OCH.sub.2CH.sub.2O),3.480-3.460(dd,1H,3-CH),3.357(s,3H,OCH.sub.3),3.181-3.156(m,1H,NCH.sub.2CHOCH.sub.2),2.796-2.760(d,1H,NCH.sub.2CHOCH.sub.2),2.750-2.689(m,2H,NCH.sub.2CH.sub.2,NCH.sub.2CHOCH.sub.2),4.485-2.396(dd,1H,6-CH.sub.2),2.249-2.176(t,1H,NCH.sub.2CH.sub.2),1.976(s,3H,COCH.sub.3),1.903-1. 883 (dd,1H,6-CH.sub.2),1.850-1.796(m,1H,NCH.sub.2CH.sub.2CH.sub.2),1.555-1.540(m,2H, NCH.sub.2CH.sub.2CH.sub.2,NCH.sub.2CH.sub.2CH.sub.2),1.444(s,9H,C(CH.sub.3).sub.3), 1.380-1.372(m,1H,NCH.sub.2CH.sub.2CH.sub.2).

(55) ESI-MS m/z: 456.2(M+H).sup.+.

(56) e) Preparation of (3R,4R,5S)-4-acetamido-5-amino-3-((3R)-3-(2-methoxylethoxyl)piperidin)-1-cyclohexene-1-carboxylic acid trifluoroacetate.

(57) ##STR00014##

(58) (3R,4R,5S)-4-acetamido-5-amino-3-((3R)-3-(2-methoxylethoxyl)piperidin)-1-cyclohexene-1-carboxylic acid trifluoroacetate was prepared with reference to Example 1 e).

(59) The characterization analysis of the product:

(60) .sup.1H-NMR(400 MHz,D.sub.2O):ppm6.942(dd,1H,2-CH),4.689-4.642(dd,1H,4-CH), 4.463-4.445(dd,1H,3-CH),3.995-3.899(dd,1H,5-CH),3.678-3.645(t,2H,OCH.sub.2CH.sub.2O),3.692-3.663(m,1H,NCH.sub.2CHOCH.sub.2),3.602-3.588(t,2H,OCH.sub.2CH.sub.2O),3.562-3.532(m,4H,NCH.sub.2CHOCH.sub.2,NCH.sub.2CH.sub.2),3.401(s,3H,OCH.sub.3),3.121-3.108(dd,2H,6-CH.sub.2),2.645-2.285(m,1H,NCH.sub.2CH.sub.2CH.sub.2),2.301-2.178(m,2H,NCH.sub.2CH.sub.2CH.sub.2,NCH.sub.2CH.sub.2CH.sub.2),2.146(s,3H,COCH.sub.3),1.876-1.850(m,1H, NCH.sub.2CH.sub.2CH.sub.2).

(61) ESI-MS m/z: 356.2(M+H).sup.+.

EXAMPLE 3

(62) a) Preparation of ethyl (3R,4S,5S)-4-acetamido-5-azido-3-((3S)-3-(2-hydroxyethoxyl)piperidin)-1-cyclohexene-1-carboxylate.

(63) ##STR00015##

(64) Ethyl (3R,4S,5S)-4-acetamido-5-azido-3-((3S)-3-(2-hydroxyethoxyl)piperidin)-1-cyclohexene-1-carboxylate was prepared with reference to t Example 1 a).

(65) The characterization analysis of the product:

(66) .sup.1H-NMR(400 MHz,CDCl.sub.3):ppm6.869(s,1H,NH),5.561-5.539(dd,1H,2-CH), 4.255-4.193(q,2H,COOCH.sub.2CH.sub.3),4.014-3.940(dd,1H,4-CH),3.834-3.193(m,2H,OCH.sub.2CH.sub.2OH),3.702-3.635(m,2H,OCH.sub.2CH.sub.2OH),3.419-3.395(dd,1H,3-CH),2.958-2.943(m,1H,NCH.sub.2CH),2.914-2. 900(m,1H,NCH.sub.2CH),2.787-2.750(m,1H,NCH.sub.2CH.sub.2),2.711-2.661(m,1H,NCH.sub.2CH.sub.2),2.585-2.261(m,1H,NCH.sub.2CH),2.554-2.534(m,1H,5-CH),2.332-2.262(m,1H,6-CH.sub.2),2.071(s,3H,COCH.sub.3),1.846-1.811(m,2H,NCH.sub.2CH.sub.2CH.sub.2),1.507-1.466(m,2H, NCH.sub.2CH.sub.2CH.sub.2),1.329-1.295(t,3H, COOCH.sub.2CH.sub.3).

(67) ESI-MS m/z: 396.2 (M+H).sup.+.

(68) b) Preparation of ethyl (3R,4S,5S)-4-acetamido-5-amino-3-((3S)-3-(2-hydroxyethoxyl)piperidin)-1-cyclohexene-1-carboxylate.

(69) ##STR00016##

(70) Ethyl (3R,4S,5S)-4-acetamido-5-amino-3-((3S)-3-(2-hydroxyethoxyl)piperidin)-1-cyclohexene-1-carboxylate was prepared with reference to Example 1 b). The characterization analysis of the product: ESI-MS m/z: 370.2 (M+H).sup.+.

(71) c) Preparation of ethyl (3R,4R,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3S)-3-(2-hydroxyethoxyl)piperidin)-1-cyclohexene-1-carboxylate.

(72) ##STR00017##

(73) Ethyl (3R,4R,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3S)-3-(2-hydroxyethoxyl)piperidin)-1-cyclohexene-1-carboxylate was prepared with reference to Example 1 c). The characterization analysis of the product: ESI-MS m/z for the product: 470.2 (M+H).sup.+.

(74) d) Preparation of (3R,4R,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3S)-3-(2-hydroxyethoxyl)piperidin)-1-cyclohexene-1-carboxylic acid.

(75) ##STR00018##

(76) (3R,4R,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3S)-3-(2-hydroxyethoxyl)piperidin)-1-cyclohexene-1-carboxylic acid was prepared with reference to Example 1 d). The characterization analysis of the product: 442.2 (M+H).sup.+.

(77) e) Preparation of (3R,4R,5S)-4-acetamido-5-amino-3-((3S)-3-(2-hydroxyethoxyl)piperidin)-1-cyclohexene-1-carboxylic acid trifluoroacetate.

(78) ##STR00019##

(79) (3R,4R,5S)-4-acetamido-5-amino-3-((3S)-3-(2-hydroxyethoxyl)piperidin)-1-cyclohexene-1-carboxylic acid trifluoroacetate was prepared with reference to Example 1 e).

(80) The characterization analysis of the product:

(81) .sup.1H-NMR(400 MHz, CDCl):ppm6.993(dd,1H,2-CH),4.401-4.354(dd,1H,4-CH), 4.027-3.981(dd, 1H,3-CH),3.612-3.462(m,2H,OCH.sub.2CH.sub.2OH),3.352-3.343(m,2H,OCH.sub.2CH.sub.2OH),3.289-3.165(m,1H,5-CH),3.046-3.019(m,1H,NCH.sub.2CH),2.952-2.898(m,2H,NCH.sub.2CH,NCH.sub.2CH.sub.2),2.856-2.804(m,1H,NCH.sub.2CH.sub.2),2.763-2.649(m,1H,NCH.sub.2CH),2.465-2.381(m,2H,6-CH.sub.2),2.082(s,3H,COCH.sub.3),1.861-1.831(m,1H,NCH.sub.2CH.sub.2CH.sub.2),1.726-1.705(m,1H,NCH.sub.2CH.sub.2CH.sub.2),1.616-1.559(m,1H,NCH.sub.2CH.sub.2CH.sub.2),1.312-1.286(m,1H,NCH.sub.2CH.sub.2CH.sub.2).

(82) ESI-MS m/z: 342.2 (M+H).sup.+.

EXAMPLE 4

(83) a) Preparation of ethyl (3R,4S,5S)-4-acetamido-5-azido-3-((3S)-3-(2-ethoxylethoxyl)piperidin)-1-cyclohexene-1-carboxylate.

(84) ##STR00020##

(85) Ethyl (3R,4S,5S)-4-acetamido-5-azido-3-((3S)-3-(2-ethoxylethoxyl)piperidin)-1-cyclohexene-1-carboxylate was prepared with reference to Example 1 a). The characterization analysis of the product: ESI-MS m/z: 448.2 (M+H).sup.+.

(86) b) Preparation of ethyl (3R,4S,5S)-4-acetamido-5-amino-3-((3S)-3-(2-ethoxylethoxyl)piperidin)-1-cyclohexene-1-carboxylate.

(87) ##STR00021##

(88) Ethyl (3R,4S,5S)-4-acetamido-5-amino-3-((3S)-3-(2-ethoxylethoxyl)piperidin)-1-cyclohexene-1-carboxylate was prepared with reference to Example 1 b). The characterization analysis of the product: ESI-MS m/z: 398.3 (M+H).sup.+.

(89) c) Preparation of ethyl (3R,4S,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3S)-3-(2-ethoxylethoxyl)piperidin)-1-cyclohexene-1-carboxylate.

(90) ##STR00022##

(91) Ethyl (3R,4S,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3S)-3-(2-ethoxylethoxyl)piperidin)-1-cyclohexene-1-carboxylate was prepared with reference to Example 1 c). The characterization analysis of the product: ESI-MS m/z: 522.2 (M+H).sup.+.

(92) d) Preparation of (3R,4S,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3S)-3-(2-ethoxylethoxyl)piperidin)-1-cyclohexene-1-carboxylic acid.

(93) ##STR00023##

(94) (3R,4S,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3S)-3-(2-ethoxylethoxyl)piperidin)-1-cyclohexene-1-carboxylic acid was prepared with reference to Example 1 d).

(95) The characterization analysis of the product:

(96) .sup.1-H-NMR(400 MHz,CDCl.sub.3):ppm6.892(dd,1H,2-CH),5.421-5.335(dd,1H,4-CH), 4.165-4.046(dd,1H,5-CH),3.769-3.745(m,1H,3-CH),5.542-3.483(m,6H,OCH.sub.2CH.sub.2O,CH.sub.2OCH.sub.2CH3),3.392-3.856(m,1H,NCH.sub.2CH),2.932-2.865(m,1H,NCH.sub.2CH),2.812-2.745(m,2H,NCH.sub.2CH.sub.2),2.545-5.465(m, 1H,6-CH.sub.2),2.389-2.275(m,1H,NCH.sub.2CH),2.254-2.178(m,1H,6-CH.sub.2),1.985(s,3H,COCH.sub.3),1.954-1.938(m,1H,NCH.sub.2CH.sub.2CH.sub.2),1.713-1.645(m,2H,NCH.sub.2CH.sub.2CH.sub.2,NCH.sub.2CH.sub.2CH.sub.2),1.420(s,9H,C(CH.sub.3).sub.3),1.398-1.252(m,1H,NCH.sub.2CH.sub.2CH.sub.2),1.191-1.156(t,3H, CH.sub.2OCH.sub.2CH.sub.3).

(97) ESI-MS m/z for the product: 470.3 (M+H).sup.+.

(98) e) Preparation of (3R,4S,5S)-4-acetamido-5-amino-3-((3S)-3-(2-ethoxylethoxyl)piperidin)-1-cyclohexene-1-carboxylic acid trifluoroacetate.

(99) ##STR00024##

(100) (3R,4S,5S)-4-acetamido-5-amino-3-((3S)-3-(2-ethoxylethoxyl)piperidin)-1-cyclohexene-1-carboxylic acid trifluoroacetate was prepared with reference to Example 1 e).

(101) The characterization analysis of the product:

(102) .sup.1H-NMR(400 MHz,CDCl.sub.3):ppm7.002(dd,1H,2-CH),4.498-4.389(dd,1H,4-CH), 3.746-3.694(m,1H,3-CH),3.625-3.534(m,4H.sub.2OCH.sub.2CH.sub.2O),3.398-3.376(m,2H,CH.sub.2OCH.sub.2CH.sub.3),3.201-3.194(dd, 1H,5-CH),3.078-3.054(m, 1H,NCH.sub.2CH),3.042-3.201(m,1H,NCH.sub.2CH),2.964-2.945(m,2H,NCH.sub.2CH.sub.2),2.879-2.850(m,1H,6-CH.sub.2),2.539-2.502(m,1H,NCH.sub.2CH),2.105-2.098(m,1H,6-CH.sub.2),2.090(s,3H,COCH.sub.3),1.879-1.786(m,2H,NCH.sub.2CH.sub.2CH.sub.2,NCH.sub.2CH.sub.2CH2),1.726-1.649(m,2H,NCH.sub.2CH.sub.2CH.sub.2,NCH.sub.2CH.sub.2CH.sub.2),1.219-1.184(t,3H, CH.sub.2OCH.sub.2CH.sub.3) ESI-MS m/z: 370.3 (M+H).sup.+.

EXAMPLE 5

(103) a) Preparation of ethyl (3R,4S,5S)-4-acetamido-5-azido-3-((3S)-3-((2-(cyclopropylmethoxyl)ethoxyl)piperidin)-1-cyclohexene-1-carboxylate.

(104) ##STR00025##

(105) Ethyl (3R,4S,5S)-4-acetamido-5-azido-3-((3S)-3-((2-(cyclopropylmethoxyl)ethoxyl)piperidin)-1-cyclohexene-1-carboxylate was prepared with reference to Example 1 a).

(106) The characterization analysis of the product: ESI-MS m/z: 450.3 (M+H).sup.+.

(107) b) Preparation of ethyl (3R,4S,5S)-4-acetamido-5-amino-3-((3S)-3- ((2-(cyclopropylmethoxyl)ethoxyl)piperidin)-1-cyclohexene-1-carboxylate.

(108) ##STR00026##

(109) Ethyl (3R,4S,5S)-4-acetamido-5-amino-3-((3S)-3-((2-(cyclopropylmethoxyl)ethoxyl)piperidin)-1-cyclohexene-1-carboxylate was prepared with reference to Example 1 b).

(110) The characterization analysis of the product:

(111) .sup.1H-NMR(400 MHz,CDCl.sub.3):ppm6.898(s,1H,NH),5.497-5.446(dd,1H,2-CH), 4.214-4.169(q,2H,COOCH.sub.2CH.sub.3),3.868-3.795(dd,1H,4-CH),3.401-3.369(m,1H,3-CH),3.321-3.286(m,2H, OCH.sub.2CH),3.256-3.222(m,1H,5-CH),2.894-2.841(m,4H, OCH.sub.2CH.sub.2O),2.706-2.676(m,1H, NCH.sub.2CH),2.492-2.444(m,1H NCH.sub.2CH),2.165-2.112(m,2H, NCH.sub.2CH,NCH.sub.2CH.sub.2),2.109-2.078((m,1H,6-CH.sub.2),2.055(s,3H,COCH.sub.3),2.018-1.998(m,1H,6-CH.sub.2),1.995-1.936(m,1H,NCH.sub.2CH.sub.2),1.751-1.688(m,1H,NCH.sub.2CH.sub.2CH.sub.2),1.494-1.428(m,1H,NCH.sub.2CH.sub.2CH.sub.2),1.355-1.321(t,3H,COOCH.sub.2CH.sub.3),1.298-1.156(m,2H,NCH.sub.2CH.sub.2CH.sub.2,NCH.sub.2CH.sub.2CH.sub.2),1.190-0.955(m,1H,CH(CH.sub.2CH.sub.2)),0.568-0.478(m,2H,CH(CH.sub.2CH.sub.2)),0.321-0.276(m,2H, CH(CH.sub.2CH.sub.2))

(112) ESI-MS m/z: 424.3 (M+H).sup.+.

(113) c) Preparation of ethyl (3R,4S,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3S)-3-((2-(cyclopropylmethoxyl)ethoxyl)piperidin)-1-cyclohexene-1-carboxylate.

(114) ##STR00027##

(115) Ethyl (3R,4S,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3S)-3-((2-(cyclopropylmethoxyl)ethoxyl)piperidin)-1-cyclohexene-1-carboxylate was prepared with reference to Example 1 c).

(116) The characterization analysis of the product: ESI-MS m/z: 524.3 (M+H).sup.+.

(117) d) Preparation of (3R,4S,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3S)-3-((2-(cyclopropylmethoxy)ethoxyl)piperidin)-1-cyclohexene-1-carboxylic acid.

(118) ##STR00028##

(119) (3R,4S,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3S)-3-((2-(cyclopropylmethoxyl)ethoxyl)piperidin)-1-cyclohexene-1-carboxylic acid was prepared with reference to Example 1 d).

(120) The characterization analysis of the product:

(121) .sup.1H-NMR(400 MHz,CDCl.sub.3):ppm6.887(dd,1H,2-CH),5.446-5.426(dd,1H,4-CH), 4.211-4.189(m,1H,5-CH),3.946-3.932(m,1H,3-CH),3.794-3.782(m,2H,OCH.sub.2CH.sub.2O),3.869-3.659(m,2H,OCH.sub.2CH.sub.2O),3.336-3.320(m,2H,OCH.sub.2CH),3.623-3.116(m,1H,NCH.sub.2CH),3.099-3.036(m,2H,NCH.sub.2CH,NCH.sub.2CH.sub.2),2.926-2.886(m,1H,NCH.sub.2CH),2.846-2.827(m,1H,6-CH.sub.2),2.389-3.313(m,1H,NCH.sub.2CH),2.073-2.066(m,1H,6-CH.sub.2),2.039-2.017(s,3H,COCH.sub.3),1.956-1.935(m,1H,NCH.sub.2CH.sub.2CH.sub.2),1.726-1.635(m,2H,NCH.sub.2CH.sub.2CH.sub.2,NCH.sub.2CH.sub.2CH2),1.421(s,9H,C(CH.sub.3).sub.3),1.864-1.031(m,1H,NCH.sub.2CH.sub.2CH.sub.2),0.899-0.836(m,1H,CH(CH.sub.2CH.sub.2)),0.527-0.507(m,2H,CH(CH.sub.2CH.sub.2)),0.215-0.204(m,2H,CH(CH.sub.2CH.sub.2)).

(122) ESI-MS m/z: 494.2 (M+H).sup.+.

(123) e) Preparation of (3R,4S,5S)-4-acetamido-5-amino-3-((3S)-3-((2-(cyclopropylmethoxyl)ethoxyl)piperidin)-1-cyclohexene-1-carboxylic acid trifluoroacetate.

(124) ##STR00029##

(125) (3R,4S,5S)-4-acetamido-5-amino-3-((3S)-3-((2-(cyclopropylmethoxy)ethoxyl)piperidin)-1-cyclohexene-1-carboxylic acid trifluoroacetate was prepared with reference to Example 1 e).

(126) The characterization analysis of the product:

(127) .sup.1H-NMR(400 MHz,MeOD):ppm6.996(dd,1H,2-CH),4.403-4.354(dd,1H,4-CH), 4.089-4.021(m,1H,3-CH),3.721-3.675(m,2H,OCH.sub.2CH.sub.2O),3.601-3.516(m,2H,OCH.sub.2CH.sub.2O),3.458-3.976(m,1H,5-CH),3.388-3.380(m,2H,OCH.sub.2CH),3.357-3.349(m,1H,NCH.sub.2CH),3.340-3.310(m,2H,NCH.sub.2CH,NCH.sub.2CH.sub.2),3.287-3.176(m,1H,NCH.sub.2CH),3.123-3.087(m,1H,6-CH.sub.2),3.047-3.015(m,1H,NCH.sub.2CH),2.524-2.455(m,1H,6-CH.sub.2),2.086(s,3H,COCH.sub.3),2.047-2.018(m,1H,NCH.sub.2CH.sub.2CH.sub.2),1.876-1.816(m,1H,NCH.sub.2CH.sub.2CH.sub.2),1.689-1.615(m,2H,NCH.sub.2CH.sub.2CH.sub.2,NCH.sub.2CH.sub.2CH.sub.2),1.056-1.027(m,1H,CH(CH.sub.2CH.sub.2)),0.549-0.518(m,2H,CH(CH.sub.2CH.sub.2)),0.243-0.219(m,2H,CH(CH.sub.2CH.sub.2)).

(128) ESI-MS m/z: 394.2 (M+H).sup.+.

EXAMPLE 6

(129) a) Preparation of ethyl (3R,4S,5S)-4-acetamido-5-azido-3-((3S)-3-((2-(methoxylethoxyl)methyl)piperidin)-1-cyclohexene-1-carboxylate.

(130) ##STR00030##

(131) Ethyl (3R,4S,5S)-4-acetamido-5-azido-3-((3S)-3-((2-(methoxylethoxyl)methyl)piperidin)-1-cyclohexene-1-carboxylate was prepared with reference to Example 1 a).

(132) The characterization analysis of the above product: ESI-MS m/z: 448.2(M+H)+.

(133) b) Preparation of ethyl (3R,4S,5S) -4-acetamido-5-amino-3-((3S)-3-((2-(methoxylethoxyl)methyl)piperidin)-1-cyclohexene-1-carboxylate.

(134) ##STR00031##

(135) Ethyl (3R,4S,5S)-4-acetamido-5-amino-3-((3S)-3-((2-(methoxyethoxyl)methyl)piperidin)-1-cyclohexene-1-carboxylate was prepared with reference to Example 1 b).

(136) The characterization analysis of the product:

(137) .sup.1H-NMR(400 MHz,CDCl.sub.3):ppm6.880(s,1H,NH),5.763-5.742(dd,1H,2-CH), 4.200-4.138(m,2H,COOCH.sub.2CH.sub.3),4.125-4.089(dd,1H,4-CH),3.805-3.757(dd,1H,3-CH),3.554-3.506(m,4H,OCH.sub.2CH.sub.2O),3.352(s,3H,CH.sub.2OCH.sub.3),3.328-3.316(m,1H,5-CH),3.282-3.219(m,2H,CH.sub.2OCH.sub.2CH.sub.2),2.874-2.812(m,2H,NCH.sub.2CH.sub.2),2.757-2.724(m,2H,NCH.sub.2CH,6-CH.sub.2),2.359.2.309(m,1H,NCH.sub.2CH),2.170-2.121(m,1H,6-CH.sub.2),2.097(s,3H,COCH.sub.3),1.967-1.856(m,1H,NCH.sub.2CH),1.623-1.565(m,2H,NCH.sub.2CH.sub.2CH.sub.2,NCH.sub.2CH.sub.2CH.sub.2),1. 422-1.336(m,1H,NCH.sub.2CH.sub.2CH.sub.2),1.278-1.215(t,3H,COOCH.sub.2CH.sub.3),1.021-0.916(m,1H,NCH.sub.2CH.sub.2CH.sub.2).

(138) ESI-MS m/z: 398.2 (M+H).sup.+.

(139) c) Preparation of ethyl (3R,4S,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3S)-3-((2-(methoxylethoxyl)methyl)piperidin)-1-cyclohexene-1-carboxylate.

(140) ##STR00032##

(141) Ethyl (3R,4S,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3S)-3-((2-(methoxylethoxyl)methyl)piperidin)-1-cyclohexene-1-carboxylate was prepared with reference to Example 1 c).

(142) The characterization analysis of the product: ESI-MS m/z: 498.2 (M+H).sup.+.

(143) d) Preparation of (3R,4S,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3S)-3-((2-(methoxylethoxyl)methyl)piperidin)-1-cyclohexene-1-carboxylic acid.

(144) ##STR00033##

(145) (3R,4S,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3S)-3-((2-(methoxylethoxyl)methyl)piperidin)-1-cyclohexene-1-carboxylic acid was prepared with reference to Example 1 d).

(146) The characterization analysis of the product:

(147) .sup.1H-NMR(400 MHz,MeOD):ppm6.847(dd,1H,2-CH),5.406-5.372(dd,1H,4-CH), 4.144-4.091(m,1H,5-CH),3.743-3.728(m,1H,3-CH),3.543-3.370(m,4H,OCH.sub.2CH.sub.2O), 3.370(s,3H,CH.sub.2OCH.sub.3),3.324-3.283 (m,1H,CH.sub.2OCH.sub.2CH.sub.2),3.056-2.905 (m,1H,CH.sub.2OCH.sub.2CH.sub.2),2.895-2.873(m,1H,NCH.sub.2CH.sub.2),2.861-2.821(m,1H,NCH.sub.2CH.sub.2),2.589-2.457(m,1H,NCH.sub.2CH),2.436-2.356(m,1H, 6-CH.sub.2),2.199-2.186(m,1H,NCH.sub.2CH), 2.125-2.072(m,1H, 6-CH.sub.2),1.978(s,3H,COCH.sub.3),1.721-1.686(m,1H,NCH.sub.2CH),1.654-1.556(m,2H,NCH.sub.2CH.sub.2CH.sub.2,NCH.sub.2CH.sub.2CH.sub.2),1.415(s,9H,C(CH.sub.3)3),1.315-1.271(m,1H,NCH.sub.2CH.sub.2CH.sub.2),1.071-1.050(m,1H,NCH.sub.2CH.sub.2CH.sub.2).

(148) ESI-MS m/z for the product: 470.2 (M+H).sup.+.

(149) e) Preparation of (3R,4S,5S)-4-acetamido-5-amino-3-((3S)-3-((2-(methoxyethoxyl)methyl)piperidin)-1-cyclohexene-1-carboxylic acid trifluoroacetate.

(150) ##STR00034##

(151) (3R,4S,5S)-4-acetamido-5-amino-3-((3S)-3-((2-(methoxyethoxyl)methyl)piperidin)-1-cyclohexene-1-carboxylic acid trifluoroacetate was prepared with reference to Example 1 e).

(152) The characterization analysis of the product:

(153) .sup.1H-NMR(400 MHz,MeOD):ppm6.969(dd,1H,2-CH),4.469-4.419(dd,1H,4-CH), 4.290-4.286(m,1H,3-CH),3.598-3.512(m,4H.sub.2OCH.sub.2CH.sub.2O),3.495-3.461(m,2H,CH.sub.2OCH.sub.2CH.sub.2),3.359(s,3H,CH.sub.2OCH.sub.3),3.082-3.070(m,1H,5-CH),3.039-3.025(m,1H,NCH.sub.2CH.sub.2),2.886-2.765 (m,1H,NCH.sub.2CH.sub.2),2.529-2.457(m,1H,NCH.sub.2CH),2.445-2.436(m,1H,6-CH.sub.2),2.286-2.175(m,1H,NCH.sub.2CH),2.138-2.086(m, 1H,6-CH.sub.2),1.992(s,3H,COCH.sub.3),1.821-1.765(m,1H,NCH.sub.2CH),1.721-1.665(m,2H,NCH.sub.2CH.sub.2CH.sub.2,NCH.sub.2CH.sub.2CH.sub.2),1.421-1.335(m,1H,NCH.sub.2CH.sub.2CH.sub.2),1.213-1.159(m,1H,NCH.sub.2CH.sub.2CH.sub.2).

(154) ESI-MS m/z: 370.2(M+H).sup.+.

EXAMPLE 7

(155) a) Preparation of ethyl (3R,4S,5S)-4-acetamido-5-azido-3-((3S)-3-((2-(ethoxylcarbonyl)amino)ethoxyl)piperidin)-1-cyclohexene-1-carboxylate.

(156) ##STR00035##

(157) Ethyl (3R,4S,5S)-4-acetamido-5-azido-3-((3S)-3-((2-(ethoxylcarbonyl)amino)ethoxyl)piperidin)-1-cyclohexene-1-carboxylate was prepared with reference to Example 1 a). The characterization analysis of the product: ESI-MS m/z: 467.2 (M+H).sup.+.

(158) b) Preparation of ethyl (3R,4S,5S) -4-acetamido-5-amino-3-((3S)-3-((2-(ethoxylcarbonyl)amino)ethoxyl)piperidin)-1-cyclohexene-1-carboxylate.

(159) ##STR00036##

(160) Ethyl (3R,4S,5S)-4-acetamido-5-amino-3-((3S)-3-((2-(ethoxylcarbonyl)amino)ethoxyl)piperidin)-1-cyclohexene-1-carboxylate was prepared with reference to Example 1 b).

(161) The characterization analysis of the product: ESI-MS m/z: 441.2 (M+H).sup.+.

(162) c) Preparation of ethyl (3R,4S,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3S)-3-((2-(ethoxylcarbonyl)amino)ethoxyl)piperidin)-1-cyclohexene-1-carboxylate.

(163) ##STR00037##

(164) Ethyl (3R,4S,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3S)-3-((2-(ethoxylcarbonyl)amino)ethoxyl)piperidin)-1-cyclohexene-1-carboxylate was prepared with reference to Example 1 c).

(165) The characterization analysis of the product: ESI-MS m/z: 541.2 (M+H).sup.+.

(166) d) Preparation of (3R,4S,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3S)-3-((2-(ethoxylcarbonyl)amino)ethoxyl)piperidin)-1-cyclohexene-1-carboxylic acid.

(167) ##STR00038##

(168) (3R,4S,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3S)-3-((2-(ethoxylcarbonyl)amino)ethoxyl)piperidin)-1-cyclohexene-1-carboxylic acid was prepared with reference to Example 1 d).

(169) The characterization analysis of the product:

(170) .sup.1H-NMR(400 MHz,CDCl.sub.3):ppm6.942(dd,1H,2-CH),5.456(m,1H,4-CH), 5.356(dd,1H,5-CH),4.145-4.008(m,2H,NHCOOCH.sub.2CH.sub.3),3.795-3.785(m,1H,3-CH),3.770-3.712(m,2H,OCH.sub.2CH.sub.2NH),3.486-3.398(m,2H,OCH.sub.2CH.sub.2NH),2.936-2.908(m,1H,NCH.sub.2CH),2.791-2.700(m,2H,NCH.sub.2CH.sub.2,NCH.sub.2CH),2.487-2.467(m,1H,NCH.sub.2CH.sub.2),2.351-2.331(m,1H,NCH.sub.2CH),2.241-2.204(m,1H,6-CH.sub.2),2.175-2.168(m,1H,6-CH.sub.2),1.962(s,3H,COCH.sub.3),1.627-1.578(m,1H,NCH.sub.2CH.sub.2CH.sub.2),1.564-1.536(m,2H,NCH.sub.2CH.sub.2CH.sub.2),1.521-1.489(m,1H,NCH.sub.2CH.sub.2CH.sub.2),1.422(s,9H,C(CH.sub.3).sub.3),1.255-1.202(m,3H,NHCOOCH.sub.2CH.sub.3).

(171) ESI-MS m/z: 513.2 (M+H).sup.+.

(172) e) Preparation of (3R,4S,5S)-4-acetamido-5-amino-3-((3S)-3-((2-(ethoxylcarbonyl)amino)ethoxyl)piperidin)-1-cyclohexene-1-carboxylic acid trifluoroacetate.

(173) ##STR00039##

(174) (3R,4S,5S)-4-acetamido-5-amino-3-((3S)-3-((2-(ethoxylcarbonyl)amino)ethoxyl)piperidin)-1-cyclohexene-1-carboxylic acid trifluoroacetate was prepared with reference to Example 1 e).

(175) The characterization analysis of the product:

(176) .sup.1H-NMR(400 MHz,MeOD):ppm6.934(dd,1H,2-CH),4.360(m,1H,4-CH), 4.104-4.052(m,2H,NHCOOCH.sub.2CH.sub.3),3.805-3.789(m,1H,3-CH),3.783-3.756(m,2H,OCH.sub.2CH.sub.2NH),3.558-3.550(dd,1H,5-CH),3.496-3.389(m,2H.sub.2OCH.sub.2CH.sub.2NH),2.969-2.926(m,1H,NCH.sub.2CH),2.805-2.756(m,2H,NCH.sub.2CH.sub.2,NCH.sub.2CH),2.528-2.501(m,1H,NCH.sub.2CH.sub.2),2.412-2.385(m,1H,NCH.sub.2CH),2.352-2.220(m,1H,6-CH.sub.2),2.196-2.163(m,1H,6-CH.sub.2),1.984(s,3H,COCH.sub.3),1.638-1.572(m,1H,NCH.sub.2CH.sub.2CH.sub.2),1.561-1.535(m,2H,NCH.sub.2CH.sub.2CH.sub.2),1.502-1.476(m,1H,NCH.sub.2CH.sub.2CH.sub.2), 1.309-1.264 (m,3H,NHCOOCH.sub.2CH.sub.3).

(177) ESI-MS m/z: 413.2 (M+H).sup.+.

EXAMPLE 8

(178) a) Preparation of ethyl (3R,4S,5S)-4-acetamido-5-azido-3-((3S)-3-(n-butoxymethyl)piperidin)-1-cyclohexene-1-carboxylate.

(179) ##STR00040##

(180) Ethyl (3R,4S,5S)-4-acetamido-5-azido-3-((3S)-3-(n-butoxymethyl) piperidin)-1-cyclohexene-1-carboxylate was prepared with reference to Example 1 a).

(181) The characterization analysis of the product: ESI-MS m/z: 422.3 (M+H).sup.+.

(182) b) Preparation of ethyl (3R,4S,5S) -4-acetamido-5-amino-3-((3S)-3-(n-butoxymethyl)piperidin)-1-cyclohexene-1-carboxylate.

(183) ##STR00041##

(184) Ethyl (3R,4S,5S)-4-acetamido-5-amino-3-((3S)-3-(n-butoxymethyl) piperidin)-1-cyclohexene-1-carboxylate was prepared with reference to Example 1 b). The characterization analysis of the product: ESI-MS m/z: 396.2 (M+H).sup.+.

(185) c) Preparation of ethyl (3R,4S,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3S)-3-(n-butoxymethyl) piperidin)-1-cyclohexene-1-carboxylate.

(186) ##STR00042##

(187) Ethyl (3R,4S,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3S)-3-(n-butoxymethyl)piperidin)-1-cyclohexene-1-carboxylate was prepared with reference to Example 1 c). The characterization analysis of the product: ESI-MS m/z: 496.2 (M+H).sup.+.

(188) d) Preparation of (3R,4S,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3S)-3-(n-butoxymethyl) piperidin)-1-cyclohexene-1-carboxylic acid.

(189) ##STR00043##

(190) (3R,4S,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3S)-3-(n-butoxymethyl)piperidin)-1-cyclohexene-1-carboxylic acid was prepared with reference to Example 1 d).

(191) The characterization analysis of the product:

(192) .sup.1H-NMR(400 MHz,CDCl.sub.3):ppm6.869(dd,1H,2-CH),5.297(dd,1H,4-CH), 4.144-4.120(dd, 1H,5-CH),3.758-3.739(dd,1H,3-CH),3. 689-3.612(m,1H,CHCH.sub.2OCH.sub.2),3.412-3.346(q,2H, OCH.sub.2CH.sub.2),3.329-3.286(m,1H,CHCH.sub.2OCH.sub.2),2.958-2.912(m,2H,NCH.sub.2CH.sub.2),2.495-2.483(m,1H,NCH.sub.2CH),2.401-2.389(m,1H,6-CH.sub.2),2.251-2.203(m,1H,NCH.sub.2CH),2.044-2.035(m,1H,6-CH.sub.2),1.988(s,3H,COCH.sub.3),1.856-1. 803(m,1H,NCH.sub.2CH),1.717-1.681(m,.sub.3H,NCH.sub.2CH.sub.2CH.sub.2,CH.sub.2CH.sub.2CH.sub.3),1.568-1.501(m,3H,NCH.sub.2CH.sub.2CH.sub.2,CH.sub.2CH.sub.2CH.sub.3),1.422(s,9H,C(CH.sub.3).sub.3),1.379-3.286(m,2H,CH.sub.2CH.sub.2CH.sub.3),0.928-0.816(t,3H,CH.sub.2CH.sub.2CH.sub.3).

(193) ESI-MS m/z: 468.2 (M+H).sup.+.

(194) e) Preparation of (3R,4S,5S)-4-acetamido-5-amino-3-((3S)-3-(n-butoxymethyl) piperidin)-1-cyclohexene-1-carboxylic acid trifluoroacetate.

(195) ##STR00044##

(196) (3R,4S,5S)-4-acetamido-5-amino-3-((3S)-3-(n-butoxymethyl) piperidin)-1-cyclohexene-1-carboxylic acid trifluoroacetate was prepared with reference to Example 1 e).

(197) The characterization analysis of the product:

(198) .sup.1-H-NMR(400 MHz,D.sub.2O):ppm 6.962(dd,1H,2-CH),4.555-4.401(dd,1H,4-CH), 4.235-4.225(dd,1H,3-CH),3.601-3.515(m,1H,CHCH.sub.2OCH.sub.2),3.498-3.490(dd,1H,5-CH),3.458-3.401(q,2H,OCH.sub.2CH.sub.2),3.368-3.312(m,1H,CHCH.sub.2OCH.sub.2),2.986-2.916(m,2H,NCH.sub.2CH.sub.2),2.504-2.479(m,1H,NCH.sub.2CH),2.434-2.392(m,1H,6-CH.sub.2),2.286-2.249(m,1H,NCH.sub.2CH),2.105-2.089(m,1H,6-CH.sub.2),1.962(s,3H,COCH.sub.3),1.871-1.825(m,1H,NCH.sub.2CH),1.735-1.699(m,3H,NCH.sub.2CH.sub.2CH.sub.2,CH.sub.2CH.sub.2CH.sub.3),1.584-1.499(m,3H,NCH.sub.2CH.sub.2CH.sub.2,CH.sub.2CH.sub.2CH.sub.3), 1.385-3.274(m,2H,CH.sub.2CH.sub.2CH.sub.3),0.999-0.826(t,3H,CH.sub.2CH.sub.2CH.sub.3).

(199) ESI-MS m/z: 368.2 (M+H).sup.+.

EXAMPLE 9

(200) a) Preparation of ethyl (3R,4S,5S)-4-acetamido-5-azido-3-((3S)-3-(2-((dimethylcarbamoyl) oxy)ethyoxyl)piperidin)-1-cyclohexene-1-carboxylate.

(201) ##STR00045##

(202) Ethyl (3R,4S,5S)-4-acetamido-5-azido-3-((3S)-3-(2-((dimethylcarbamoyl)oxy)ethyoxyl)piperidin)-1-cyclohexene-1-carboxylate was prepared with reference to Example 1 a).

(203) The characterization analysis of the product:

(204) .sup.1H-NMR(400 MHz,CDCl.sub.3):ppm6.857(s,1H,NH),5.577-5.555(dd,1H,2-CH), 4.662-4.617(m,1H,4-CH),4.286-4.179(m,2H,COOCH.sub.2CH.sub.3),4.059-3.869(m,2H,CH.sub.2OCON),3.706-3.638(m,1H,3-CH),3.529-3.485(m,2H,OCH.sub.2CH.sub.2OCO),2.912-2.887(s,6H,N(CH.sub.3).sub.2),2.826-2.816(m,2H,NCH.sub.2CH, NCH.sub.2CH.sub.2CH.sub.2),2.724-2.696(m,1H,NCH.sub.2CH),2.593-2.546(m,1H,NCH.sub.2CH.sub.2CH.sub.2),2.327-2.230(m,2H,NCH.sub.2CH,5-CH),2.046-2.015(s,3H,COCH.sub.3),1.952-1.913(m,1H,6-CH.sub.2),1.712-1.653(m,2H,6-CH.sub.2,NCH.sub.2CH.sub.2CH.sub.2),1.512-1.409(m,2H,NCH.sub.2CH.sub.2CH.sub.2),1.386-1.355(m,1H,NCH.sub.2CH.sub.2CH.sub.2),1.346-1.298(t,3H,COOCH.sub.2CH.sub.3).

(205) ESI-MS m/z: 467.3 (M+H).sup.+.

(206) Preparation of ethyl (3R,4S,5S) -4-acetamido-5-amino-3-((3S)-3-(2-((dimethylcarbamoyl)oxy)ethyoxyl)piperidin)-1-cyclohexene-1-carboxylate.

(207) ##STR00046##

(208) Ethyl (3R,4S,5S)-4-acetamido-5-amino-3-((3S)-3-(2-((dimethylcarbamoyl)oxy)ethyoxyl)piperidin)-1-cyclohexene-1-carboxylate was prepared with reference to Example 1 b).

(209) The characterization analysis of the product: ESI-MS m/z: 441.2 (M+H).sup.+.

(210) c) Preparation of ethyl (3R,4S,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3S)-3-(2-((dimethylcarbamoyl) oxy)ethyoxyl)piperidin)-1-cyclohexene-1-carboxylate.

(211) ##STR00047##

(212) Ethyl (3R,4S,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3S)-3-(2-((dimethylcarbamoyl)oxy)ethyoxyl)piperidin)-1-cyclohexene-1-carboxylate was prepared with reference to Example 1 c). The characterization analysis of the product: ESI-MS m/z: 541.2 (M+H).sup.+.

(213) d) Preparation of (3R,4S,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3S)-3-(2-((dimethylcarbamoyl)oxy)ethyoxyl)piperidin)-1-cyclohexene-1-carboxylic acid.

(214) ##STR00048##

(215) (3R,4S,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3S)-3-(2-((dimethylcarbamoyl)oxy)ethyoxyl)piperidin)-1-cyclohexene-1-carboxylic acid was prepared with reference to Example 1 d).

(216) The characterization analysis of the product:

(217) .sup.1-H-NMR(400 MHz,MeOD):ppm6.824(m,1H,2-CH),4.840-4.586(m,1H,4-CH), 4.008-3.956(m,1H,5-CH),3.683-3.593(m,2H,CH.sub.2OCON),3.495-3.472(m,1H,3-CH),2.946-2.898(s,6H,N(CH.sub.3).sub.2),2.890-2.857(m,2H,OCH.sub.2CH.sub.2OCO),2.807-2.780(m,1H,NCH.sub.2CH),2.728-2.717(m,1H,NCH.sub.2CH),2.685-2.673(m,1H,NCH.sub.2CH.sub.2CH.sub.2),2.630-2.584(m,1H,NCH.sub.2CH.sub.2CH.sub.2),2.434-3.390(m,1H,NCH.sub.2CH),2.214-2.144(m,2H,6-CH.sub.2),1.989(s,3H,COCH.sub.3),1.957-1.986(m,2H,NCH.sub.2CH.sub.2CH.sub.2),1.740-1.722(m,1H,NCH.sub.2CH.sub.2CH.sub.2),1.434(s,9H,C(CH.sub.3).sub.3).

(218) ESI-MS m/z: 513.3 (M+H).sup.+.

(219) e) Preparation of (3R,4S,5S)-4-acetamido-5-amino-3-((3S)-3-(2-((dimethylcarbamoyl)oxy)ethyoxyl)piperidin)-1-cyclohexene-1-carboxylic acid trifluoroacetate.

(220) ##STR00049##

(221) (3R,4S,5S)-4-acetamido-5-amino-3-((3S)-3-(2-((dimethylcarbamoyl)oxy)ethyoxyl)piperidin)-1-cyclohexene-1-carboxylic acid was prepared with reference to Example 1 e).

(222) The characterization analysis of the product:

(223) .sup.1H-NMR(400 MHz,MeOD):ppm7.001(m,1H,2-CH),4.560-4.555(m,1H,4-CH), 4.291-4.268(m,2H,CH.sub.2OCON),3.639-3.603(m,1H,3-CH),3.565-3.508(m,2H,OCH.sub.2CH.sub.2OCO),3.337-3.333(s,6H,N(CH.sub.3).sub.2),3.289-3.275(m,1H,5-CH),3.106-3.007(m,1H,NCH.sub.2CH),2.956-2.863(m,1H,NCH.sub.2CH),2.731-2.699(m,1H,NCH.sub.2CH.sub.2CH.sub.2),2.654-2.487(m,1H,NCH.sub.2CH.sub.2CH.sub.2),2.455-2.346(m,1H,NCH.sub.2CH),2.289-2.607(m,2H,6-CH.sub.2),2.058(s,3HCOCH.sub.3),1.969-1.946(m,2H,NCH.sub.2CH.sub.2CH.sub.2),1.768-1.652(m,1H,NCH.sub.2CH.sub.2CH.sub.2).

(224) ESI-MS m/z: 413.3 (M+H).sup.+.

EXAMPLE 10

(225) a) Preparation of ethyl (3R,4S,5S)-4-acetamido-5-azido-3-((3S)-3-(2-((morpholine-4-carbonyl)oxy) ethyoxyl)piperidin)-1-cyclohexene-1-carboxylate.

(226) ##STR00050##

(227) Ethyl (3R,4S,5S)-4-acetamido-5-azido-3-((3S)-3-(2-((morpholine-4-carbonyl)oxy)ethyoxyl)piperidin)-1-cyclohexene-1-carboxylate was prepared with reference to Example 1 a).

(228) The characterization analysis of the product:

(229) .sup.1H-NMR(400 MHz,CDCl.sub.3):ppm6.844(s,1H,NH),5.471-5.449(dd,1H,2-CH), 4.710-4.654(m,1H,4-CH),4.258-4.171(q,2H,COOCH.sub.2CH.sub.3),4.036-3.962(m,2H,CH.sub.2OCON),3.692-3.664(m,4H,2xNCH.sub.2CH.sub.2O),3.456-3.433(m,6H,2xNCH.sub.2CH.sub.2O,OCH.sub.2CH.sub.2O),2. 899-2.883(m,1H,NCH.sub.2CH),2.880-2.873(m,2H,NCH.sub.2CH,NCH.sub.2CH.sub.2CH.sub.2),2.730-2.703(m,1H,NCH.sub.2CH.sub.2CH.sub.2),2.594-2.546(m,1H,NCH.sub.2CH),2.324-2.296(m,1H,5-CH),2.272-2.229(m,1H,6-CH.sub.2),2.047(s,3H,COCH.sub.3),1.993-1.938(m,1H,6-CH.sub.2),1.697-1.643(m,2H, NCH.sub.2CH.sub.2CH.sub.2),1.492-1.421(m,2H, NCH.sub.2CH.sub.2CH.sub.2),1.392-1.304(t,3H, COOCH.sub.2CH.sub.3).

(230) ESI-MS m/z: 509.3 (M+H).sup.+.

(231) b) Preparation of ethyl (3R,4S,5S) -4-acetamido-5-amino-3-((3S)-3-(2-((morpholine-4-carbonyl)oxy)ethyoxyl)piperidin)-1-cyclohexene-1-carboxylate.

(232) ##STR00051##

(233) Ethyl (3R,4S,5S)-4-acetamido-5-amino-3-((3S)-3-(2-((morpholine-4-carbonyl)oxy)ethyoxyl)piperidin)-1-cyclohexene-1-carboxylate was prepared with reference to Example 1 b). The characterization analysis of the product: ESI-MS m/z: 483.2 (M+H).sup.+.

(234) c) Preparation of ethyl (3R,4S,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3S)-3-(2-((morpholine-4-carbonyl)oxy)ethyoxyl)piperidin)-1-cyclohexene-1-carboxylate.

(235) ##STR00052##

(236) Ethyl (3R,4S,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3S)-3-(2-((morpholine-4-carbonyl)oxy)ethyoxyl)piperidin)-1-cyclohexene-1-carboxylate was prepared with reference to Example 1 c). The characterization analysis of the product: ESI-MS m/z: 583.3 (M+H).sup.+.

(237) d) Preparation of (3R,4S,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3S)-3-(2-((morpholine-4-carbonyl)oxy)ethyoxyl)piperidin)-1-cyclohexene-1-carboxylic acid.

(238) ##STR00053##

(239) (3R,4S,5S)-4-acetamido-5-((tert-butyloxycarbonyl) amino)-3-((3S)-3-(2-((morpholine-4-carbonyl)oxy)ethyoxyl)piperidin)-1-cyclohexene-1-carboxylic acid was prepared with reference to Example 1 d).

(240) The characterization analysis of the product: ESI-MS m/z: 555.2 (M+H).sup.+.

(241) e) Preparation of (3R,4S,5S)-4-acetamido-5-amino-3-((3S)-3-(2-((morpholine-4-carbonyl)oxy)ethyoxyl)piperidin)-1-cyclohexene-1-carboxylic acid trifluoroacetate.

(242) ##STR00054##

(243) (3R,4S,5S)-4-acetamido-5-amino-3-((3S)-3-(2-((morpholine-4-carbonyl)oxy) ethyoxyl)piperidin)-1-cyclohexene-1-carboxylic acid trifluoroacetate was prepared with reference to Example 1 e).

(244) The characterization analysis of the product:

(245) .sup.1H-NMR(400 MHz,MeOD):ppm6.993(dd,1H,2-CH),4.335-4.286(m,2H, CH.sub.2OCON), 4.245-4.223(m,1H,4-CH),3.914-3.897(m,1H,5-CH),3.663-3.604(m,4H, 2xNCH.sub.2CH.sub.2O),3.856-3.450(m,6H,2xNCH.sub.2CH.sub.2O,OCH.sub.2CH.sub.2O),3.102-3.054(m,1H,NCH.sub.2CH),3.002-2.921(m,2H,NCH.sub.2CH,NCH.sub.2CH.sub.2CH.sub.2),2.912-2.886(m,1H,NCH.sub.2CH.sub.2CH.sub.2),2.765-2.744(m,1H,NCH.sub.2CH),2.736-2.720(m,1H,6-CH.sub.2),2.512-2.443(m,1H,6-CH.sub.2),2.085(s,3H,COCH.sub.3),1.947-1.927(m,2H, NCH.sub.2CH.sub.2CH.sub.2),1.664-1.652(m,2H, NCH.sub.2CH.sub.2CH.sub.2).

(246) ESI-MS m/z: 455.2 (M+H).sup.+.

EXAMPLE 11

(247) a) Preparation of ethyl(3R,4R,5S)-4-acetamido-5-(2,3-bis(tert-butoxycarbonyl)guanidino)-3-((3S)-3-(2-methoxylethyoxyl)piperidin)-1-cyclohexene-1-carboxylate.

(248) ##STR00055##

(249) According to the above equation, 0.5 mmol of ethyl(3R,4R,5S)-4-acetamido-5-amino-3-((3S)-3-(2-methoxylethyoxyl)piperidin)-1-cyclohexene-1-carboxylate was added into a single neck flask, and 10 mL of acetonitrile was added and stirred at room temperature. Then 1.55 mmol of DIPEA and 0.5 mmol of N, N-bis-BOC-1H-1-guanidinopyrazole was added, and the mixture was kept stirring at room temperature for 3 hours. Once TLC (PE:EA=1:1) indicated the reaction was completed, 20 ml of water was added, and the mixture was extracted with EA, dried with anhydrous sodium sulfate and evaporated to dryness. After being purified by the column (PE:EA=3:1), a white solid was obtained.

(250) The characterization analysis of the product:

(251) .sup.1H-NMR(400 MHz,CDCl.sub.3):ppm6.909(s,1H,NHAc),6.273-6.253(m,1H,2-CH), 4.345-4.286(m,1H,4-CH),4.211-4.198(q,2H,COOCH.sub.2CH.sub.3),4.188-4.111(m,1H,3-CH),3.659-3.570(m,2H,OCH.sub.2CH.sub.2OCH.sub.3),3.525-3.503(m,2H,OCH.sub.2CH.sub.2OCH.sub.3),3.378(s,3H,OCH.sub.2CH.sub.2OCH.sub.3),3.327-3.315(m,1H,5-CH),2.918-2.866(m,1H,NCH.sub.2CH),2.824-2.800(m,2H,NCH.sub.2CH,NCH.sub.2CH.sub.2CH.sub.2),2.525-2.476(m,1H,NCH.sub.2CH.sub.2CH.sub.2),2.315-2.283(m,1H,NCH.sub.2CH),2.179-2.126(m,1H,6-CH),2. 041-2.022(m,1H,6-CH),1.998(s,3H,COCH.sub.3),1.991-1.661(m,3H,NCH.sub.2CH.sub.2CH.sub.2),1.493(s,18H,2xC(CH.sub.3).sub.3),1.432-1.425(m,1H,NCH.sub.2CH.sub.2CH.sub.2),1.312-1.286(t,3H,COOCH.sub.2CH.sub.3).

(252) ESI-MS m/z: 626.43 (M+H)+.

(253) b) Preparation of ethyl (3R,4R,5S) -4-acetamido-5-(2,3-bis(tert-butoxycarbonyl)guanidino)-3-((3S)-3-(2-methoxylethyoxyl)piperidin)-1-cyclohexene-1-carboxylic acid.

(254) ##STR00056##

(255) 0.368 mmol of ethyl (3R,4R,5S)-4-acetamido-5-(2,3-bis(tert-butoxycarbonyl)guanidino)-3-((3S)-3-(2-methoxylethyoxyl)piperidin)-1-cyclohexene-1-carboxylate was added into a single neck flask, and 5.5 ml of 1,4-dioxane, 0.55 mol of water, and 0.55 ml of 1N KOH aqueous solution were then added and stirred overnight at room temperature. Once TLC (PE:EA=1:1) indicated the reaction was completed, the solvent was evaporated to dryness through an oil pump, 5 ml of methanol was then added for dissolution. Acid resin was used for adjusting pH to 5, the mixture was filtered, evaporated to dryness, and eluted by column purification (DCM:MeOH=10:1). A white solid was obtained finally.

(256) The characterization analysis of the product:

(257) .sup.1H-NMR(400 MHz,CDCl.sub.3):ppm6.949(s,1H,NHAc),4.323-4.304(m,1H,2-CH), 4.188-4.167(m,1H,4-CH),3.635-3.628(m,1H,3-CH),3.622-3.585(m,2H,OCH.sub.2CH.sub.2OCH.sub.3),3.528-3.518(m,2H,OCH.sub.2CH.sub.2OCH.sub.3),3.489-3.465(m,1H,5-CH),3.561-3.440(m,1H,NCH.sub.2CH),3.388-3.372(s,3H,OCH.sub.2CH.sub.2OCH.sub.3),3.016-2.908(m,1H,NCH.sub.2CH),2. 878-2.864(m,1H,NCH.sub.2CH.sub.2CH.sub.2),2.557-2.525 (m,1H,NCH.sub.2CH.sub.2CH.sub.2),2.324-2.294(m,1H,NCH.sub.2CH),2.286-2.224(m,1H,6-CH),1.993-1.956(m,1H,6-CH),1.924(s,3H,COCH.sub.3),1.716-1.695(m,1H,NCH.sub.2CH.sub.2CH.sub.2),1.612-1.535(m,2H,NCH.sub.2CH.sub.2CH.sub.2),1.484(s,18H,2xC(CH.sub.3).sub.3),1.321-1.255(m,1H, NCH.sub.2CH.sub.2CH.sub.2).

(258) ESI-MS m/z: 598.3 (M+H)+.sup.+.

(259) c) Preparation of (3R,4R,5S)-4-acetamido-5-guanidino-3-((3S)-(2-methoxylethyoxyl)piperidin)-1-cyclohexene-1-carboxylic acid trifluoroacetate.

(260) ##STR00057##

(261) (3R,4S,5S)-4-acetamido-5-5-(2,3-bis(tert-butoxycarbonyl)guanidino)-3-((3S)-3-(2-methoxylethyoxyl)piperidin)-1-cyclohexene-1-carboxylic acid was added into a single neck flask, and the solution of 50% DCM in TFA was then added and stirred at room temperature for 1 hour. Once TLC (DCM:MeOH=10:1) indicated the reaction was completed, the solvent was evaporated to dryness and the excess TFA was drawn out through an oil pump. Diethyl ether was added for trituration. After filtering, a white solid product was obtained.

(262) The characterization analysis of the product:

(263) .sup.1H-NMR(400 MHz,MeOD):ppm6.973(m,1H,2-CH),4.385-4.368(m,1H,4-CH), 3.947-3.934(m,1H,3-CH),3.922-3.906(m,1H,5-CH),3.724-3.653(m,2H,OCH.sub.2CH.sub.2OCH.sub.3),3.633-3.626(m,1H,NCH.sub.2CH),3.597-3.588(m,2H.sub.2OCH.sub.2CH.sub.2OCH.sub.3),3.512-3.478(m,1H,NCH.sub.2CH),3.414(s,3H,OCH.sub.2CH.sub.2OCH.sub.3),3.161-3.131(m,1H,NCH.sub.2CH.sub.2CH.sub.2),3.034-3.021(m,1H,NCH.sub.2CH.sub.2CH.sub.2),2.898-2. 877(m,1H,NCH.sub.2CH),2.479-2.407(m,1H,6-CH),2.289-2.156(m,1H,6-CH),2.074(s,3H,COCH.sub.3),1.988-1.876(m,2H,NCH.sub.2CH.sub.2CH.sub.2),1.828-1.793(m,2H,NCH.sub.2CH.sub.2CH.sub.2).

(264) ESI-MS m/z for the product: 398.3 (M+H).sup.+.

EXAMPLE 12

(265) a) Preparation of ethyl(3R,4R,5S)-4-acetamido-5-(2,3-bis(tert-butoxycarbonyl)guanidino)-3-((3R)-3-(2-methoxylethyoxyl)piperidin)-1-cyclohexene-1-carboxylate.

(266) ##STR00058##

(267) Ethyl(3R,4R,5S)-4-acetamido-5-(2,3-bis (tert-butoxycarbonyl)guanidino)-3-((3R)-3-(2-methoxylethyoxyl)piperidin)-1-cyclohexene-1-carboxylate was prepared according to Example 11 a).

(268) The characterization analysis of the product:

(269) .sup.1H-NMR(400 MHz,CDCl.sub.3):ppm6.98(s,1H,NHAc),6.278-6.263(m,1H,2-CH), 4.356-4.292(m,1H,4-CH),4.219-4.204(q,2H,COOCH.sub.2CH.sub.3),4.195-4.125(m,1H,3-CH),3.670-3.589(m,2H,OCH.sub.2CH.sub.2OCH.sub.3),3.560-3.521(m,2H,OCH.sub.2CH.sub.2OCH.sub.3),3. 381(s,3H,OCH.sub.2CH.sub.2OCH.sub.3),3.337-3.322(m,1H,5-CH),2.915-2.862(m,1H,NCH.sub.2CH),2.828-2.805(m,2H,NCH.sub.2CH,NCH.sub.2CH.sub.2CH.sub.2),2.558-2.492(m,1H,NCH.sub.2CH.sub.2CH.sub.2),2.321-2.292(m,1H,NCH.sub.2CH),2.186-2.122(m,1H, 6-CH),2.098-2.057(m,1H,6-CH),1.997(s,3H,COCH.sub.3),1.990-1.668(m,3H,NCH.sub.2CH.sub.2CH.sub.2),1.495(s,18H,2xC(CH.sub.3).sub.3),1.435-1.427(m,1H,NCH.sub.2CH.sub.2CH.sub.2),1.315-1.283(t,3H,COOCH.sub.2CH3).

(270) ESI-MS m/z: 626.4 (M+H).sup.+.

(271) b) Preparation of (3R,4R,5S)-4-acetamido-5-(2,3-bis(tert-butoxycarbonyl)guanidino)-3-((3R)-3-(2-methoxylethyoxyl)piperidin)-1-cyclohexene-1-carboxylic acid.

(272) ##STR00059##

(273) (3R,4R,5S)-4-acetamido-5-(2,3-bis(tert-butoxycarbonyl)guanidino)-3-((3R)-3-(2-methoxylethyoxyl)piperidin)-1-cyclohexene-1-carboxylic acid was prepared according to Example 11 b).

(274) The characterization analysis of the product:

(275) .sup.1H-NMR(400 MHz,CDCl.sub.3):ppm6.963(s,1H,NHAc),4.353-4.311(m,1H,2-CH), 4.193-4.172(m,1H,4-CH),3.641-3.635(m,1H,3-CH),3.635-3.588(m,2H,OCH.sub.2CH.sub.2OCH.sub.3),3.535-3.522(m,2H,OCH.sub.2CH.sub.2OCH.sub.3),3.492-3.469(m, 1H,5-CH),3.573-3.450(m, 1H,NCH.sub.2CH),3.394-3.379(s,3H,OCH.sub.2CH.sub.2OCH.sub.3),3.030-2.928(m,1H,NCH.sub.2CH),2.890-2.869(m,1H,NCH.sub.2CH.sub.2CH.sub.2),2.562-2.532(m,1H,NCH.sub.2CH.sub.2CH.sub.2),2.330-2.290(m,1H,NCH.sub.2CH),2.286-2.222(m,1H,6-CH),1.998-1.967(m,1H,6-CH),1.929(s,3H,COCH.sub.3),1.720-1.698(m,1H,NCH.sub.2CH.sub.2CH.sub.2),1.623-1.530(m,2H,NCH.sub.2CH.sub.2CH.sub.2),1.491(s,18H,2xC(CH.sub.3).sub.3),1.322-1.261(m,1H, NCH.sub.2CH.sub.2CH.sub.2).

(276) ESI-MS m/z: 598.3 (M+H).sup.+.

(277) c) Preparation of (3R,4R,5S)-4-acetamido-5-guanidino-3-((3R)-3-(2-methoxylethyoxyl)piperidin)-1-cyclohexene-1-carboxylic acid trifluoroacetate.

(278) ##STR00060##

(279) (3R,4R,5S)-4-acetamido-5-guanidino-3-((3R)-3-(2-methoxylethyoxyl) piperidin)-1-cyclohexene-1-carboxylic acid trifluoroacetate was prepared with reference to the Example 11 c).

(280) The characterization analysis of the product:

(281) .sup.1H-NMR(400 MHz,MeOD):ppm6.978(m,1H,2-CH),4.389-4.372(m,1H,4-CH),3.952-3.938(m,1H,3-CH),3.929-3.903(m,1H,5-CH),3.732-3.661(m,2H,OCH.sub.2CH.sub.2OCH.sub.3),3.635-3.629(m,1H,NCH.sub.2CH),3.601-3.591(m,2H,OCH.sub.2CH.sub.2OCH.sub.3),3.519-3.476(m,1H,NCH.sub.2CH),3.421(s,3H,OCH.sub.2CH.sub.2OCH.sub.3),3.169-3.129(m,1H,NCH.sub.2CH.sub.2CH.sub.2),3.089-3.065(m,1H,NCH.sub.2CH.sub.2CH.sub.2),2.901-2.898(m,1H,NCH.sub.2CH),2.482-2.411(m,1H,6-CH),2.296-2.165(m,1H,6-CH),2.095(s,3H,COCH.sub.3),1.991-1.867(m,2H,NCH.sub.2CH.sub.2CH.sub.2),1.835-1.798(m,2H,NCH.sub.2CH.sub.2CH.sub.2).

(282) ESI-MS m/z for the product: 398.3 (M+H).sup.+.

EXAMPLE 13

(283) a) Preparation of ethyl(3R,4S,5S)-4-acetamido-5-azido-3-((3R)-3-((2-(methoxylethyoxyl)methyl)piperidin)-1-cyclohexene-1-carboxylate.

(284) ##STR00061##

(285) Ethyl(3R,4S,5S)-4-acetamido-5-azido-3-((3R)-3-((2-(methoxylethyoxyl)methyl)piperidin)-1-cyclohexene-1-carboxylate was prepared with reference to Example 1 a).

(286) The characterization analysis of the product: ESI-MS m/z: 424.2 (M+H).sup.+.

(287) b) Preparation of ethyl (3R,4S,5S)-4-acetamido-5-amino-3-((3R)-3-((2-(methoxylethyoxyl)methyl)piperidin)-1-cyclohexene-1-carboxylate.

(288) ##STR00062##

(289) Ethyl (3R,4S,5S)-4-acetamido-5-amino-3-((3R)-3-((2-(methoxylethyoxyl)methyl)piperidin)-1-cyclohexene-1-carboxylate was prepared with reference to Example 1 b).

(290) The characterization analysis of the product: ESI-MS m/z: 398.2 (M+H).sup.+.

(291) c) Preparation of ethyl(3R,4R,5S)-4-acetamido-5-(2,3-bis((tert-butyloxycarbonyl)guanidine)-3-((3R)-3-((2-(methoxylethyoxyl)methyl)piperidin)-1-cyclohexene-1-carboxylate.

(292) ##STR00063##

(293) Ethyl(3R,4R,5S)-4-acetamido-5-(2,3-bis((tert-butyloxycarbonyl)guanidine)-3-((3R)-3-((2-(methoxylethyoxyl)methyl)piperidin)-1-cyclohexene-1-carboxylate was prepared with reference to Example 11 a).

(294) The characterization analysis of the product:

(295) .sup.1H-NMR(400 MHz,CDCl.sub.3):ppm6.668-6.563(m,1H,2-CH),4.352-4.298(m,1H,4-CH),4.235-4.216(q,2H,COOCH2CH3),4.198-4.130(m,1H,3-CH),3.670-3.589(m,2H.sub.2OCH2CH2OCH3),3.560-3.535(m,4H,OCH2CH2OCH3),3.391-3.388(m,2H,CHCH2OCH2CH2OCH3)3.382-3.379(m,1H,5-CH),3.369(s,3H,OCH2CH2OCH3),2.925-2.860(m,1H,NCH2CH),2.836-2.809(m,2H,NCH2CH,NCH2CH2CH2),2.569-2.498(m,1H,NCH2CH2CH2),2.335-2.296(m,1H,NCH2CH),2.190-2.120(m,1H,6-CH),2.101-2.086(m,1H,6-CH),1.999(s,3H,COCH3),1.989-1.670(m,3H,NCH2CH2CH2),1.497(s,18H,2xC(CH3)3),1.447-1.435(m,1H,NCH2CH2CH2),1.339-1.291(t,3H,COOCH2CH3).

(296) ESI-MS m/z: 640.4 (M+H).sup.+.

(297) d) Preparation of (3R,4R,5S)-4-acetamido-5-(2,3-bis(tert-butoxycarbonyl)guanidino)-3-((3R)-3-((2-(methoxylethyoxyl)methyl)piperidin)-1-cyclohexene-1-carboxylic acid.

(298) ##STR00064##

(299) (3R,4R,5S)-4-acetamido-5-(2,3-bis(tert-butoxycarbonyl)guanidino)-3-((3R)-3-((2-(methoxylethyoxyl)methyl)piperidin)-1-cyclohexene-1-carboxylic acid was prepared with reference to the Example 11 b).

(300) The characterization analysis of the product:

(301) .sup.1H-NMR(400 MHz,CDCl.sub.3):ppm6.670-6.559(m,1H,2-CH),4.363-4.302(m,1H,4-CH),4.205-4.139(m,1H,3-CH),3.678-3.592(m,2H,OCH.sub.2CH.sub.2OCH.sub.3),3.589-3.539(m,4H,OCH.sub.2CH.sub.2OCH.sub.3),3.396-3.389(m,2H,CHCH.sub.2OCH.sub.2CH.sub.2OCH.sub.3),3.389-3.382(m,1H,5-CH)3.380(s,3H,OCH.sub.2CH.sub.2OCH.sub.3),2.929-2.875 (m,1H,NCH.sub.2CH),2.826-2.803(m,2H,NCH.sub.2CH,NCH.sub.2CH.sub.2CH.sub.2),2.573-2.506(m,1H,NCH.sub.2CH.sub.2CH.sub.2),2.345-2.306(m,1H,NCH.sub.2CH),2.198-2.133(m,1H,6-CH),2.109-2.098(m,1H,6-CH),1.997(s,3H,COCH.sub.3),1.990-1.686(m,3H,NCH.sub.2CH.sub.2CH.sub.2),1.495(s,18H,2xC(CH.sub.3).sub.3),1.452-1.430(m,1H,NCH.sub.2CH.sub.2CH.sub.2).

(302) ESI-MS m/z: 612.4 (M+H).sup.+.

(303) e) Preparation of (3R,4R,5S)-4-acetamido-5-guanidino-3-((3R)-3-((2-(methoxylethyoxyl)methyl)piperidin)-1-cyclohexene-1-carboxylic acid trifluoroacetate.

(304) ##STR00065##

(305) (3R,4R,5S)-4-acetamido-5-guanidino-3-((3R)-3-((2-(methoxylethyoxyl)methyl)piperidin)-1-cyclohexene-1-carboxylic acid trifluoroacetate was prepared with reference to the Example 11 c).

(306) The characterization analysis of the product:

(307) .sup.1H-NMR(400 MHz,MeOD):ppm6.675-6.555(m,1H,2-CH),4.368-4.312(m,1H,4-CH),4.265-4.239(m,1H,3-CH),3.698-3.605(m,2H,OCH.sub.2CH.sub.2OCH.sub.3),3.589-3.545(m,4H,OCH.sub.2CH.sub.2OCH.sub.3),3.405-3.395(m,2H,CHCH.sub.2OCH.sub.2CH.sub.2OCH.sub.3),3.390-3.380(m,1H,5-CH)3.379(s,3H,OCH.sub.2CH.sub.2OCH.sub.3),2.935-2.881(m,1H,NCH.sub.2CH),2.833-2.809(m,2H,NCH.sub.2CH,NCH.sub.2CH.sub.2CH.sub.2),2.586-2.526(m,1H,NCH.sub.2CH.sub.2CH.sub.2),2.352-2.316(m,1H,NCH.sub.2CH),2.201-2.140(m,1H,6-CH),2.120-2.105 (m,1H,6-CH),1.998(s,3H,COCH.sub.3),1.991-1.696(m,3H,NCH.sub.2CH.sub.2CH.sub.2),1.465-1.445(m,1H,NCH.sub.2CH.sub.2CH.sub.2).

(308) ESI-MS m/z: 412.4 (M+H).sup.+.

EXAMPLE 14

(309) a) Preparation of ethyl(3R,4R,5S)-4-acetamido-5-(2,3-bis(tert-butoxycarbonyl)guanidino)-3-((3S)-3-(2-ethyoxylmethyl)piperidin)-1-cyclohexene-1-carboxylate.

(310) ##STR00066##

(311) Ethyl(3R,4R,5S)-4-acetamido-5-(2,3-bis(tert-butoxycarbonyl)guanidino)-3-((3S)-3-(2-ethyoxylmethyl)piperidin)-1-cyclohexene-1-carboxylate was prepared with reference to Example 11 a).

(312) The characterization analysis of the product:

(313) .sup.1H-NMR(400 MHz,CDCl.sub.3):ppm6.979(s,1H,NHAc),6.384-6.365(m,1H,2-CH), 4.421-4.409(m,1H,4-CH),4.223-4.205(q,2H,COOCH.sub.2CH.sub.3),4.005-3.988(q,2H,CH.sub.2OCH.sub.2CH.sub.3),3.821-3.756(m,1H,3-CH),3.512-3.489(m,1H,5-CH),3.435-3.371(m,1H,CH.sub.2OCH.sub.2CH.sub.3),3.338-3.246(m,1H,CH.sub.2OCH.sub.2CH.sub.3),2.612-2.548(m,1H,NCH.sub.2CH.sub.2CH.sub.2),2.453-2.439(m,1H,NCH.sub.2CH.sub.2CH.sub.2),2.357-2.291(m,2H,6-CH.sub.2,NCH.sub.2CH),2.152-2.073(m,2H, 6-CH.sub.2, NCH.sub.2CH),1.988(s,3H,COCH.sub.3),1.895-1.837(m,1H,NCH.sub.2CH),1.612-1.586(m,1H,NCH.sub.2CH.sub.2CH.sub.2),1.541-1.512(m,2H,NCH.sub.2CH.sub.2CH.sub.2),1.505-1.496(m,1H,NCH.sub.2CH.sub.2CH.sub.2),1.490(s,9H,C(CH.sub.3).sub.3),1.485(s, 9H,C(CH.sub.3).sub.3),1.325-1.295(t,3H,COOCH.sub.2CH.sub.3),1.121-1.109(t,3H, CH.sub.2OCH.sub.2CH.sub.3).

(314) ESI-MS m/z: 610.4 (M+H).sup.+.

(315) b) Preparation of (3R,4R,5S)-4-acetamido-5-(2,3-bis(tert-butoxycarbonyl)guanidino)-3-((3S)-3-(2-ethyoxylmethyl)piperidin)-1-cyclohexene-1-carboxylic acid.

(316) ##STR00067##

(317) (3R,4R,5S)-4-acetamido-5-(2,3-bis(tert-butoxycarbonyl)guanidino)-3-((3S)-3-(2-ethyoxylmethyl)piperidin)-1-cyclohexene-1-carboxylic acid was prepared with reference to Example 11 b).

(318) The characterization analysis of the product:

(319) .sup.1-H-NMR(400 MHz,MeOD):ppm6.797(m,1H,2-CH),4.339-4.312(m,1H,4-CH), 3.856-3.798(m,1H,3-CH),3.524-3.472(q,2H,CH.sub.2OCH.sub.2CH.sub.3),3.418-3.395(m,1H,CH.sub.2OCH.sub.2CH.sub.3),3.756-3.371(m,1H,5-CH),3.338-3.246(m,1H,CH.sub.2OCH.sub.2CH.sub.3),3.179-3.154(m,1H,NCH.sub.2CH.sub.2CH.sub.2),3.117-3.091(m,1H,NCH.sub.2CH.sub.2CH.sub.2),2.987-2.947(m,1H,NCH.sub.2CH),2.693-2.641(m,1H,6-CH.sub.2),2.595-2.541(m,1H,NCH.sub.2CH),2.362-2.304(m,1H,6-CH.sub.2),2.056-2.005(m,1H,NCH.sub.2CH),1.982(s,3H,COCH.sub.3),1.828-1.741(m,2H,NCH.sub.2CH.sub.2CH.sub.2),1.645-1.614(m,2H,NCH.sub.2CH.sub.2CH.sub.2),1.555(s,9H,C(CH.sub.3).sub.3),1.490(s,9H,C(CH.sub.3).sub.3),1.214-1.156(t,3H, CH.sub.2OCH.sub.2CH.sub.3).

(320) ESI-MS m/z: 582.3 (M+H)+.

(321) c) Preparation of (3R,4R,5S)-4-acetamido-5-guanidino-3-((3S)-3-(2-ethyoxylmethyl)piperidin)-1-cyclohexene-1-carboxylic acid trifluoroacetate.

(322) ##STR00068##

(323) (3R,4R,5S)-4-acetamido-5-guanidino-3-((3S)-3-(2-ethyoxylmethyl)piperidin)-1-cyclohexene-1-carboxylic acid trifluoroacetate was prepared with reference to Example 11 c).

(324) The characterization analysis of the product:

(325) .sup.1H-NMR(400 MHz,D.sub.2O):ppm6.854(m,1H,2-CH),4.468-4.454 (q,2H,CH.sub.2OCH.sub.2CH.sub.3),3.949-3.938(m,1H,4-CH),3.639-3.604(m,1H,3-CH),3.582-3.564(q,2H,CH.sub.2OCH.sub.2CH.sub.3),3.547-3.539(m,1H,CH.sub.2OCH.sub.2CH.sub.3),3.529-3.518(m,1H,5-CH),3.514-3.507(m,1H,CH.sub.2OCH.sub.2CH.sub.3)3.489-3.465(m,1H,NCH.sub.2CH.sub.2CH.sub.2),3.425-3.404(m,1H,NCH.sub.2CH.sub.2CH.sub.2),3.382-3.371(m,1H,NCH.sub.2CH),3.120-3.151(m,1H,6-CH.sub.2),2.970-2.926(m,1H,NCH.sub.2CH),2.521-2.496(m,1H,6-CH.sub.2),2.449-2.346(m,1H,NCH.sub.2CH),2.081(s,3H,COCH.sub.3),2.028-1.981(m,1H,NCH.sub.2CH.sub.2CH.sub.2),1.885-1.779(m,2H,NCH.sub.2CH.sub.2CH.sub.2),1.268-1.236(m,1H,NCH.sub.2CH.sub.2CH.sub.2), 1.195-1.161(t,3H, CH.sub.2OCH.sub.2CH.sub.3).

(326) ESI-MS m/z: 382.3 (M+H).sup.+.

Contrast Example 1

(327) (3R,4R,5S)-4-acetamido-5-amino-3-((3S)-3-(2-ethyoxylmethyl)piperidin)-1-cyclohexene-1-carboxylic acid trifluoroacetate was prepared with reference to the method used in above Example 1, and its structure was shown hereinafter.

(328) The characterization analysis of the product:

(329) .sup.1H-NMR(400 MHz,D.sub.2O):ppm6.970(dd,1H,2-CH),4.476-4.425(dd,1H,4-CH), 4.325-4.267(dd,1H,5-CH),3.694-3.586(dd,1H,3-CH),3.597-3.471(m,4H,OCH.sub.2CH.sub.3,CHCH.sub.2O),3.436-3.430(dd,1H,NCH.sub.2CH.sub.2),3.089-3.056(dd,1H,NCH.sub.2CH.sub.2),3.050-3.021(dd,1H,NCH.sub.2CH),2.825-2.654(dd,1H,NCH.sub.2CH),2.532-2.463(dd,1H,6-CH.sub.2),2.086-2.195(dd,1H,6-CH.sub.2),2.086(s,3H,COCH.sub.3),1.983-1.948(m,1H,NCH.sub.2CH),1.812-1.705(m,3H,NCH.sub.2CH.sub.2CH.sub.2,NCH.sub.2CH.sub.2CH.sub.2),1.313-1.299(m,1H,NCH.sub.2CH.sub.2CH.sub.2),1.193-1.159(t,3H,OCH.sub.2CH.sub.3).

(330) ESI-MS m/z: 340.2 (M+H).sup.+.

Contrast Example 2

(331) (3R,4R,5S)-4-acetamido-5-amino-3-((3R)-3-(2-ethyoxylmethyl)piperidin)-1-cyclohexene-1-carboxylic acid trifluoroacetate was prepared with reference to the method used in above Example 1, and its structure was shown hereinafter.

(332) The characterization analysis of the product:

(333) .sup.1H-NMR(400 MHz,D.sub.2O):ppm6.975(dd,1H,2-CH),4.482-4.463(dd,1H,4-CH), 4.352-4.278(dd,1H,5-CH),3.702-3.642(dd,1H,3-CH),3.609-3.495(m,4H,OCH.sub.2CH.sub.3,CHCH.sub.2O),3.457-3.449(dd, 1H,NCH.sub.2CH.sub.2),3.102-3.087(dd,1H,NCH.sub.2CH.sub.2),3.068-3.043(dd,1H,NCH.sub.2CH),2.863-2.735(dd,1H,NCH.sub.2CH),2.612-2.489(dd,1H,6-CH.sub.2),2.108-2.095(dd,1H,6-CH.sub.2),2.003(s,3H,COCH.sub.3),1.987-1.965(m,1H,NCH.sub.2CH),1.826-1.735(m,3H,NCH.sub.2CH.sub.2CH.sub.2,NCH.sub.2CH.sub.2CH.sub.2),1.356-1.312(m,1H,NCH.sub.2CH.sub.2CH.sub.2),1.201-1.169(t,3H,OCH.sub.2CH.sub.3).

(334) ESI-MS m/z: 340.2 (M+H).sup.+.

Contrast Example 3

(335) (3R,4R,5S)-4-acetamido-5-amino-3-((3S)-3-((2,2,2-trifluoroethoxyl)methyl)piperidin)-1-cyclohexene-1-carboxylic acid trifluoroacetate was prepared with reference to the method used in above Example 1, and its structure was shown hereinafter.

(336) The characterization analysis of the product:

(337) .sup.1H-NMR(400 MHz,D.sub.2O):ppm6.975(dd,1H,2-CH),4.482-4.463(dd,1H,4-CH), 4.352-4.278(dd,1H,5-CH),3.702-3.642(dd,1H,3-CH),3.609-3.495(m,4H,OCH.sub.2CH.sub.3,CHCH.sub.2O),3.457-3.449(dd, 1H,NCH.sub.2CH.sub.2),3.102-3.087(dd,1H,NCH.sub.2CH.sub.2),3.068-3.043(dd,1H,NCH.sub.2CH),2.863-2.735(dd,1H,NCH.sub.2CH),2.612-2.489(dd,1H,6-CH.sub.2),2.108-2.095(dd,1H,6-CH.sub.2),2.003(s,3H,COCH.sub.3),1.987-1.965 (m,1H,NCH.sub.2CH),1.826-1.735(m,3H,NCH.sub.2CH.sub.2CH.sub.2,NCH.sub.2CH.sub.2CH.sub.2),1.356-1.312(m,1H,NCH.sub.2CH.sub.2CH.sub.2),1.201-1.169(t,3H,OCH.sub.2CH.sub.3).

(338) ESI-MS m/z: 340.2 (M+H).sup.+.

Experimental Examples

(339) Activity assay for influenza virus neuraminidase.

(340) Experimental materials:

(341) NA (neuraminidase) solution: allantoic fluid of chicken embryo infected by influenza virus;

(342) Enzyme catalyzed reaction system: 330 mmol/L of MES buffer solution (pH 3.5); 200 mol/L of fluorogenic substrate MUNANA (2-(4-methylumbelliferyl)--D-N-acetylneuraminic acid);

(343) 4 mmol/L of CaCl.sub.2 solution; Stop buffer: 14 mmol/L of NaOH solution (14 mmol/L, 83% ethanol); Oseltamivir: 1 mmol/L;

(344) Enzyme activity assay: NA solution (with different diluted concentration) 40 l; MES (fatty acid methyl ester sulfonate) (330 mmol/L): 10 l; CaCl.sub.2 (4 mmol/L): 10 l; MUNANA (200 mol/L): 10 l; H.sub.2O: 30 l; The above materials were mixed together and incubated for 15 minutes, and 150 l of the stop buffer was added: EX=355 nm XM=460 nm

(345) Note: The dynamic curve of fluorescence values can be detected without addition of the stop buffer, in order to verify whether the enzyme reaction system works normally.

(346) Experiment operation for screening a NA inhibitor is as follows:

(347) 1. Oseltamivir or the compounds prepared in Examples 1-12 were diluted in proportion of 1:10, 1:100, 1:1000, 1:10000.

(348) 2. Reaction steps: 30 l of NA solution and 10 l of Oseltamivir, or the compounds prepared in Examples 1-12 were well mixed and incubated for 30 minutes, and following ingredients were then added: 10 l of MES (330 mmol/L), 10 l of CaCl.sub.2(4 mmol/L), 10 l of MUNANA (200 l), and 30 l of H.sub.2O. After they were well mixed and incubated for 15 minutes, 150 l of the stop buffer was added and well mixed finally. The fluorescence values were detected: EX=355 nm XM=460 nm.

(349) 3. Data analysis: analyzed by GraphPad Prism Demo, and the assay results are shown in the table below:

(350) TABLE-US-00001 TABLE 1 Inhibiting activity for influenza virus neuraminidase H274Y MUTANT WIDE-TYPE IC.sub.50 DRUG-RESISTANT EXAMPLE COMPOUND STRUCTURE (nM) STRAIN IC.sub.50 (nM) 1 a a 2 0 b b 3 b b 4 b b 5 a b 6 b b 7 b b 8 b b 9 b b 10 c b 11 a a 12 0 b b 13 b b 14 b b POSITIVE Oseltamivir phosphate a c CONTROL CONTRAST EXAMPLE 1 b b CONTRAST EXAMPLE 2 b b CONTRAST EXAMPLE 3 b b

(351) Note: a: <5 nM; b: 5 nM-100 nM; c: 100-1000 nM. nM is nmol/L.

(352) It can be seen from the above results that the compounds disclosed in the present disclosure show great inhibiting activities on influenza viruses, and the activity will significantly increase when guanidine is located on C.sub.5 site of the compound, or the substituent in S configuration is in the 3 site of piperidine ring. In addition, the compounds of all examples show great inhibiting activities on NA of H274Y mutant viruses strains which are drug-resistant to Oseltamivir. Specifically, Examples 1 and 11 further show a perfect inhibiting activities on wild-type and H274 mutant virus strains. This shows their application prospect in terms of preparation of anti-influenza drug, and a new drug is expected.

(353) Although the present invention has been disclosed in the form of preferred embodiments and variations thereon, it will be understood that numerous additional modifications and variations could be made thereto without departing from the scope of the invention.

(354) For the sake of clarity, it is to be understood that the use of a or an throughout this application does not exclude a plurality, and comprising does not exclude other steps or elements.