CA1121358A - Antiviral amine and amidine derivatives of glycerol and propanediols - Google Patents

Abstract

ABSTRACT OF THE DISCLOSURE
Novel amine derivatives of di-O-(n-higher alkyl and alkenyl)-propanediols, and their pharmaceutically acceptable acid addition salts, are useful for combating viral infections in mammals. Of particular inter-est is 1-[2,3-di(n-hexadecyloxy)propyl]-4-aminomethyl-4-phenylpiperidine and its pharmaceutically acceptable acid addition salts.

Description

35~

P.C. 5860 A~TIVIR~L AMI~E AND A~SIDI~ DERIVATIVES
OF GLYCEROL AND PROPA~EDIOLS
_ !
Virus inf~ections which at~ack ma~mals, including man, are normally contagious afflictions ~hich are capable o~
causing great hu~an suffering and economic loss. Unfortunately, the discovery of antiviral compounds is far more complicated and difficult than the discovery of antibacterial an~ anti-fungal agents. This is due~ in part, to the close structural slmilarity o~ viruses and the structure of certain essential - ` cellular componen~s such as ribonucleic and deoxyribonucleic acids. Nevertheless, numerous nonviral "antiviral agents", i.e. substances "which can produce either a protective or i therapeutic effect to the clear detectable advantage of the ~ virus in~ected host, or any material that can si~nificantly enhance antibody ~ormation, improve antibody activity, improve non-speci~ic resistance, speed convalescence or depress symptomsn l~errman-e~. al., Proc. Soc. Exptl. Biol. Med., 103, ?
625 (1969~, have been described in the li~erature. The list of ,, i `' ,1 ~b~ I

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reported antiviral agents includes~ to name a few, interferon and I
synthetic materials such as amantadine hydrochloride, pyrimidines,' biguanides, guanidine, pteridines and methisazone. Because of j the rather narrow range of viral infections that can be treated by each of the antiviral agents commercially available at the present time, new synthetic antiviral agents are always welcomed as potentially valuable additions to the armamentarium of medical !
techno~ogy.
~' ~he cells of mammals produce, in response to virus in~ection ,!

, a substance which enables cells to resist the multiplication of a v~riety of viruses. The viral-resisting or viral-interfering substances are referred to as "interferons". The interferons are glycoproteins which may differ in their physico-chemical properties, but all exhibit the same biological properties; namely~, they inhibit a wide range of unrelated viruses, have no toxic or other dele~erious effects on cells, and are species-specific ¦
(Loc~art, Frontiers of Biologyl Vol. 2, "Interferons", edited by Finter, ~. B. Saunders Co., Philadephia, 1966, pages 19-~0).
, No practical, economical m~thod has yet ~ean developed for the preparation of exogenous interferon for routine cli~ical use !
against viral inf~ctions. An lternative approach to producing interferonhas, ~herefore, been pursued, which comprises adminis- !
tering to the animal to be pro~ected or ~reated a non-viral substance which stimulates- or induces- production of interfero~

in the cells. The inter~eron produced in this fashion is referred to as "endogenous" interferon.
U~ SD Patent No. 2,738,351 discloses that compounds of the general formula ~ . Rl-X-CH-Z-ALK-B

! R2 Y C~2 ~~

~Z~L3S8 wherein each of Rl and R2 may be alkyl, aralkyl, aryl, cyclo-alkyl, nitro-substituted aryl, halogen-substituted aryl, alkyl-substituted aryl, or alkoxy~substituted aryl, each of X, Y and Z may be oxygen, sulfur or sulfonyl, ALK is straight or branched alkylene of from one to six carbon atoms, and B may be di(lower)-alkylamino, piperidino, morpholino, pyrrolidino, (lower alkyl)-pyrrolidino, N'-alkyl-piperazino or pipecolino, are local anesthetic agents. Additionally, the discussion of alternate synthetic routes (s Col. 1, 11.57-70, of said patent) discloses intermediates of the above formula wherein B is amino and (lower alkyl)amino. However, none of the compounds specifically enum-erated in the disclosure of said patent contain an alkyl Rl or R2 larger than _-pentyl. Furthermore, in none of these compounds are both Rl and R2 alkyl and both X and Y oxygen.
Insecticidal and miticidal compounds of the formula R -CH
CH~(CH2)q~A

wherein R and R may each be inter alia, lower alkylthio; ~ is 1 2 ~
0 to 5; and A may be, inter alia, l-piperidino or di(lower alkyl) amino are dissolved in Japanese Patent J7-6042~177.
It has now been discovered that certain novel amine and amidine derivatives of di-O-(n-higher alkyl and alkenyl)-glycerols and propanediols are capable of combating viral infections in mammals. The novel compounds of this invention have the formula:

~ 2 2 CH2 ~- N ~ V
Rl-O-CH ~ = \
R -O-CH ~

and the pharmaceutically acceptable acid addition salts thereof, 3S~3 wherein Rl and R2 are each selected from the group consisting of normal alkyl of from 14 to 18 carbon atoms and octadec-9-enyl.
The invention disclosed herein comprises the novel antiviral compounds of formula V, the novel pharmaceutical compositions containing an antivirally eEfective amount of a compound oE formula V as the essential active ingredient in a pharmaceutically acceptable carrier, the novel method of prophy-lactically controlling a viral infection in a mammal which com-prises administering an amount effective to prophylactically control said viral infection of a compound of formula V, and the novel method of inducing the production of interferon in a mammal which comprises administering an amount effective to induce the production of interferon of a compound of formula V.
The compounds of this invention exhibit antiviral activity against a wide variety of viruses in vivo in mammals and ln vitro in mammalian tissue culture. At least a substantial portion of this activity results from the ability of said com-pounds to induce the production of interferon in the cells, i.e.
endogenous interferon.
By "pharmaceutically acceptable" acid addition salts is meant those salts which are non-toxic at the dosages admini-s-tered. The pharmaceutically acceptable acid addition salts which may be employed include such water-soluble and water-insoluble salts as the hydrochloride, hydrobromide, phosphate, nitrate, sulfate, acetate, hexafluorophosphate, citrate, glu-conate, benzoate, propionate, butyrate, sulfosalicylate, maleate, laurate, malate, Eumarate, succinate, oxalate, tartrate, amsonate (4,4'-diaminostilbene-2,2'-disulfonate), pamoate (l,l'-methylene-bls-2-hydroxy-3-naphthoate), stearate, 3-hydroxy-2-naphthoate, p-toluenesulfonate, methanesulfonate, lactate, and suramin salts.
One preferred group of the compounds of formula V
consists of the hydrochloride salts of the bases of formula V.

Another preferred group of the compounds of formula V
consists of those wherein Rl and P~2 are each normal alkyl of from 14 to 18 carbon atoms.
Another pre:Eerred group of the compounds of formula V
consists of those wherein Rl and R2 are each normal alkyl of from 14 to 18 carbon atoms and contain the same number of carbon atoms.
Another preferred group of the compounds of formula V
consists of those wherein Rl and R2 are each _-hexadecyl.
Particularly valuable are the following compounds and their pharmaceutically acceptable acid addition salts:
1-[2,3-di(_-octadecyloxy)propyl]-4-aminomethyl-4-phenylpiperidine, 1-[2,3-di(_-hexadecyloxy)propyl]-4-aminomethyl-4-phenylpiperidine, and 1-[2,3-di( -tetradecyloxy)propyl]-4-aminomethyl-4-phenylpiperidine.
According to the invention, the novel compounds of formula V and their pharmaceutically acceptable acid addition salts are prepared by the steps of:
(a) reducing a compound of the formula ~ CN

CH2 N ~ XII; and Rl-O CH ~
R2-0-CH2 ~

(b) where required, converting the resulting compound of formula V to a pharmaceutically acceptable acid addition salt thereof.
The compounds of formula XII above may be prepared from the appropriate 1,2-di-O-(n-h.igher alkyl or alkenyl)-gly-cerol starting materials by methods familiar to those skilled in ~ - 5 -. ~

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the art. For example, the tosyl derivative of the starting material may be reacted with 4-cyano-4-phenylpiperidine hydro-chloride.
Acid addition salts oE the bases of formula V may be prepared by conventional procedures such as by mixing the amine or amidine compound in a suitable solvent with the required acid and recovering the salt by evaporation or by precipitation upon adding a non-solvent for the salt. Hydrochloride salts may readily be prepared by passing hydrogen chloride through a solution of the amine or amidine compound in an organic solvent.
As can be seen by reference to the examples herein, many of the isolated hydrochloride or dihydrochloride salts of the bases of formulae I V tend to contain a significant water content. Whether this observed "trapped" water is randomly occluded during crystallization, or corresponds to formation of true molecular hydrates, or results from the occurrence of some other phenomenon, is not known. In any event, the salts containing "trapped" water may be efficaciously formulated and administered without pre-liminary dehydration.

, ''1 The 1,2-di-0-~n-higher alkyl)-glycerol starting materials may be prepared by the method of Kates, M. et al., Bicichemistry, ~, 394 tl963). The 1,3-di-0-(n-higher alkyl)-glycerol starting materials may ke prepared by the method of Damico, R., et al., J. Lipid ~es., 8, 63 (1967). The 1,2- and 1,3-di-0-(n-higher alkenyl)-glycerol starting materials may be prepared by the , method of Bauman, ~.J. and Mangold, H.K., J. Ori3. Chem., 31, 498 j, ~lCi66)-~ The antiviral activity of the compounds of this invention ' was determined by the use of two independent procedures. In 'i the first, the test compound is administered to mice by the intraperitoneal route eighteen to twenty-our hours prior to challenging them with a lethal dose of encephalomyocarditis ~C~
,' virus. Survival data are taken during the ten days after I challenge and compared with the data for unprotected animals.
The procedure in which the drug is given eighteen to twenty-four hours before, and at a dis~inctly diffirent site from, virus injection is desiynea to eliminate local effects between drug j and ~irus and identify only those compounds which produce a ~0 systemic antiviral response' ~n the second proceduret monolayers of human nasal polyp cells grown on microtiter plat~s are treated with the test compound akout eighteen hours before treatment with a lethal dose of vesicular stomatitis virus (VSV). The test compound is washed away fxom the monolayers be~ore virus treatment. Culture fluid extracted from the plates after a post challeng~ incubation period !
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l l is titrated for the amount of infectious virus present in microti~er ¦ plates of ~-929 mouse fibroblasts. Comparison is made with the ¦ virus yield data for culture fluid extracted from unprotected I polyp cells.
Additionally, many of the compounds of this invention were ¦ tested for their ability to enhance the known antiviral activity ¦ of polyinosinic:polycytidylic acid. Finally, certain of the compounds were also tested for their ability to induce circulatin ¦ interferon in mice after parenteral administration, using the 10 ¦ procedure described by Hoffman, W.W., et al., ~ntimicrobial ~ ~ .
¦ Agents and Chemotherapy, 3, 498-501 (1973).
¦ Parenteral, topical or intranasal administration of the above-described amines and amidines to a mammal before exposure of the mammal to an infectious virus provides rapid resistance to the virus. Preferably, admini.stration should take place from about two days to about one day before exposure to the virus, although this will vary somewhat with the particular animal speci ~s and the particular infectious virus.
When the materials of this invention are administered, they ¦ are most easily and economically used in a dispersed form in an ¦ acceptable carrier. When it is said that this material is ¦ dispersed, it means that the particles may be molecular in size ¦ and held in true solution in a suitable solvent or that the particles m be colloidal in size and dispersed through a liquid ~, L3~

.
phase in the form o a suspension or an emulsion. The term "dispersed" also means that the particles may be mixed ~ith and spread throughout a solid carrier so that the mixture is in the form of a powder or dust. This term is also meant to encompass , mixtures which are suitable for use as sprays, including sol- I
~' utions, suspensions or emulsions of the agents o~ this invention. ¦
, When administer2d parenterally (subcutaneously, intra- ?
musculaxly, intraperitoneally) the matexials of this in~ention are used at a level of from about 1 mg./kg. of body weight to 10 ; about 250 mg./kg. body weight. The favored range is from about ; 5 mg./kg. to about 100 mg./kg. of body weight, and the preferred range from about 5 mg. to about 50 mg./kg. of body weight. The dosage, of course, is dependent upon the mammal being treated I' and the particular amine or amidine compound involved and is to be determined by the individual responsible for its administratiol.
, Generally, small doses will be administered initially ~ith gradual increase in dosage until the optimal dosage level is determined for the particular subject under treatment.
1 Vehicles suitable for parenteral injection may be either aqueous such as water, isotonic saline, isotonic dexkrose, Ringer';s solution, or non-aqueous such as fatty oils of vegetable origin (cottonseed, peanut oil, corn, sesame~ and other non-aqueous ~ehicles which will not interfere with the efficacy of the prepara-tion and are non-toxic in the volume or proportion used ~glycerol, ethanol, propylene glycol, sorbitol). Additionally, compositions suitable for extemporaneous preparation of solutions pxior to administration may advantageously be made. Such compositions may incluae liquid diluents, for example, propylene glycol, diethyl carbonate, glycerol, sorbitol.
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l~Z~358 In practicing the intranasal route of administration o~ i this invention any practical method can be used to contact the antiviral agent with the respiratory tract of the ma~mal.
; Effective methods include administration of the agent by intra- i nasal or nasopharyngeal drops and by inhalation as delivered by a !
nebulizer or an aerosol. Such methods of administration are of , practical importance because they provide an easy, safe and efficient method of practicing this invention. For intranasal ' aa~inistration o~ the agent, usually in an accepta~le carrier, a !, concentration of agent between 1.0 mg./ml. and 100 mg.~ml. is ¦~ satisfactory. Concentrations in the range of about 30 to 50 j mg./ml. allow administration of a convenient volume of material.
,i For topical application the antiviral agents are most l,, conveniently used in an acceptable carrier to permit ease and 1 control of application and better absorption. Here also concen-', tration~ in the ran~e of from about 1.0 mg./ml. to about 250 '1 mg./ml. are satisfactory. In general, in the above two me~hods of , administration a dose within the range of about 1.0 mg./kg. to ahout 2~0 my./kg. of body weight and, preferably, from about 5.0 mg./kg. to about 50 mg./kg. of body weight wi11 be administered The compounds e~iploye* in this invention may be employed alone, i~e., without other medicinals~ as mixtures of more than one of the herei~-described compounds, or in combination with ' other medicinal agents, such as analgesics, anesthetics, anti-; septics, decongestants, antbiotics, vaccines, bufferin~ agents and inorganic salts, to afford desirable pharmacological prop-erties. Further, they may be administered in combination with hyaluronidase to avoid or, at least, to minimi~e local irritation' ana to ;ncrease ~he rate of absorption of the compound. Hyal- !
~0 , uronidase levels of at least about 150 (U.S.P.) units are ., . .

" effective in this respect although higher or lower levels can, of course, be used.
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Those materials of this invention which are water-insoluble, including those which are of low and/or difficult solubility in water, are, for optimum results, administered in formulations, e.~., suspensions, emulsions, which permit formation of particle ; sizes of less than about 20~. The particle sizes of the form-ulations influence their biological activity apparently through better absor~tion of the active materials. In formulating these materi.als various surface active agents ~nd protective colloids t I' are used. Suitable surface.active agents are the partial esters ,~ of common fatty acids, such as lauric~ oleic, stearic, with ~, hexitol anhydrides derived from sorbitol, and the polyoxyethylene I
derivatives of such ester products. Such products are sold under ¦
. the trademarks "Spans" and ~Tweens," respectively, and are avail-, able from ICI United States Inc., Wilmington, Del. Cellulose ethers, especially cellulose methyl ether tMethocel~ available '! from the Dow Chemical Co., Midland, Mich.) are highly efficient ~1 as protective colloids for use in emulsions containing the ' materials of this invention.
, The wa~er soluble materials described herein re adminis-tered for optimum results in aqueous solution. Typically tney are : : administered in phosphate buffered saline. The water-insoluble compounds are administered in formulations of the type described above or in various other formulations as previously noted.
Dimethylsulfoxide serves as a suitable vehicle for water-insoluble compounds. A representative formulation for such compounds ; comprises formulating 25 to 100 mg. of the chosen drug.as an emul-. . I

I sion by meltl;ng and mixing with e~ual parts of polysorbate 80 and ,, . I
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ing. Sodium chloride is added in a concentrated solution to a final concentration of 0O14 M and sodium phosphate, pH 7, is added . to a final concentration of 0~01 M to give, for example, the fol-i 5 ,' lowing representative composltion: ¦

J, mg./ml~
' Drug 50.0 , Polysorbate 80 50.0 , Glycerin 50-0 10 1~ Sodium Phosphate Monobasic Hydrous 1. 4 t ! . Sodium Chloride 7.9 ,j Water 842.0 , 1001.3 ,, In certain instances, as where clumping of the drug particles occurs, sonication is employed to provide a homo~enous ' system~ I
jl The followiny examples illustrate the invention but are not ¦
to he construed as limiting the same.
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1,3-Di-0-(n-hexadecyl)-2-0-(3-aminopropyl)-glycerol Hydrochloride A. 1,3-Di-0-(n-hexadecyl)-2-0-(2-cyanoeth~ lycerol I A mixture of 1,3-di-0-(n-hexadecyl)-glycerol (80 g., 148 S mmoles), acrylonitrile (1.49 kg., 28.1 mmoles) and aqueous 2N
sodium hydroxide ~1~2 1.) was heated to 50C. Tetrabutylammonium¦
.I hydroxide (19.2 g.. o~ 40 wt. % aqueous solution, 29.15 mmoles) '. was s~owly added, causing the temperature of the exothermic ~ reaction mixture to rise to about 80 to 90C. The.reaction ! mixture was then stirred for 20 m~nutes without any external heating, followed by cooling to 20C. and addition of water . (1.0 ~ . ~ solid material, a mixture o~ unreacted and i cyanoe~hylated 1,3-di-O~tn-hexadecyl1-glycerol, was isolated and , txeated again with fresh acrylonitrile ~1.49 kg., 28.1 mmoles), aqueous 2N sodium.hy~roxide (1.2 1.) and tetrabutylammonium hydroxide ~19~2 g. of 40 wt. ~ ~queous solution, 29.15 mmoles) for 20 minutes with stirring at 50C.~ followed by cooling and addition of water ll.0 1.~. The resulti~g 1~3-di-0-(n hexadecyl3--

2-0-(2-cyanoethyl)-glycerol solids were filtered, washed con-secu~ively with water, methanol and acetonitrile, and dried 182 g., 93% yield, mcp. 45-46C., ir (C~C13) 2250 cm 1, n.m.r.
~CDC13) ~ 3~92 (t, 2, NCCH2C~ 0-), 3~33-3.67 (m~ g, OCHICH2O~2ClSH31~2)r ~-62 ~t, 2~ NCC ~ CH~O~) and 0~75-1.58 (m, 62, aliphatic protons)J.
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B. Title Compound A mixture of 1,3-di-0-~n-hexadecyl)-2-0-~2~cyanoethyl)- i glycerol (20.5 g., 34.5 mmoles), tetrahydrofuran ~200 ml.), ethanol (10 ml.) and Raney nickel catalyst (3 g.) was saturated with ammonia gas at 0 to 5C. and then hydrogenated ~;0 psi) in a Paar hydrogenator fo~ 3 hours at room temperature. The mixture ¦
was then filtered, the catalyst washed with tetrahydrofuran (50 ml.), and the total filtrate evaporated in vacuo to an oil.
'l This procedure was repeated three more times with fresh rea~tants .
, and catalyst to yield a total of 77 g. of oil. The oil was dissolved in ether ~500 ml.) and-the solution washed with 2 wt. %
aqueous ammonium hydroxide solution (500 ml.~, dried (MgS04), filtered and evaporated in vacuo t~ yield a solid. The solid was ~, dissolved in methanol (300 ml. )~and the solution saturated with ~ hydrogen chloride gas and then e~aporated in vacuo to a sOlia.
This solid was crystallized from ethyl acetate to yield the named product with a slight impurity (63 g.l 72% yield, m.p. 69-70~C.~, and then recrystalliæed twice ~rom isopropanol: acetonitrile (1:1, 800 ml.) [47.5 g., 54% yield, m.p. ~8 S9~C., n.m.r. (CDC13) ~ 3.84 ~t, 2, X2NCH2CH2CH20-), 3.55 (m, 9, -OCH[CH2OC~C15H31]23,

3-24 ~t~ 2~ H2NCH2~H2CH2-), 2-~4 (m, 2, H2NCH2CH2CH20-) and j;
, 0.90-1.32 (m9 62t aliphatic protons), elemental analysis calculated: 72.04% C; 12.73~ H; 2.21% N; found: 7~.80% C; 12,41~ ¦
~; 2.30~
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; EXAMPLES 2-7 In like manner to that described.in Example 1 the following compounds were prepared by using the appropriate 1,3- or 1,2-di-O-[n-higher alkyl)-glycerol as starting material:

Rl-O-fH2 IH-o~c~l2~3NH2 .1 , R2-0-CEI2 . '', ' .
f~2- ~CH2, 3NH2 ~ O-~H II

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35l~ i 1,3-Di-0-(n-hexadecyl)-2-0-(2-aminoethyl)-glycerol H~drochloride A. 1,3 di-0-(n-hexadecyl)-2-0-(2-carboxyethyl)-glycerol . A mixture of 1,3-di-0-~n-hexadecyl) 2-0-~2-cyanoethyl)-S . glycerol t4.8 g., 8.1 mmoles), concentrated hydrochloric acid . (~0 ml.) and formic acid ~50 ml.) was stirred for 16 hours at reflux, then cooled and extracted with ether (3 X 100 ml.). The . combined ether extract was washed with water ~200 ml.), dried ~MgSO4), ~iltered and evaporated in vacuo to yield 1,3-di-O~
! ln-hexadecyl~-2-0 ~2-carboxyethyl)-glycerol solids ~4.5 g.~, ', which were purified by silica gel chromatography ~elution with toluene:ethanol)~3.5 g., 71% yield, m.p. 43-45C., ir (CHC13) i 1740 cm 1~ n.m.r. ~CDC13) S 3.93 (t, J = 6 ~z, 2, -OC~ CH2COOH) and 2.6~ (t, J = 6 Hz, 2, -OC~2CH2COOH33.
.. B. Title ~ompound ~ ¦
: 1,3 Di-0-(n-~exadecyl)-2-0-~2-carboxyethyl)-glycerol (3.5 g., ~.7 mmoles) was dissolved in a mixture of benzene (55 ml.) and concentrated sulfuric acid (5.89 g.). Hydrazoic cid (Ç.34 ml. o~l

4.65 wt. ~ benzene solutlon, 6.0 mmoles) was then added dropwise 20 ; and ~he resulting mixture stirred for 2 hours at room temperature.
Thin layer chromatography (TLC) analysis showed about 50% reaction o~ the 2-carboxye~h~l ~ompound. Pdditional hydrazoic acid ~6.34 ml. of 4 9 65 wt. % benzene solution, 6.0 mmoles) was added dropwise and the reaction mixture stirred for another 16 hours at 40C. T1C analysis now showed that the reaction was essentially complete. ~ater (50 ml.) and aqueous 2~ sodium hydroxide were then added and thP resulting mixture extracted with ether i i , I
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1, t3 X 200 ml.). The combined ether extract was dried (Na2S04~, filtered, saturated with hydrogen chloride gas and evaporated in vacuo to yield a solid. The solid was purified by silica gel ! chromatography telution with c~loroform:methanol) and recrystal-; lized from hot ethyl acetate [570 mg.l 16% yield, m.p. 79-80C., n-m-r. tCDC133 ~ 3~95 ~m, 2, -OCH2CH2NH2) and 3.22 (m, 2, ~ ~OCH2CH?NH2), elemental analysis calculated: 71.62~ C;
'i 12.67% H; 2.Z6% N; f~und: 70.90% C; 12.19% H; ~.05% Njl.
' EXAMPLE 9 '1,3-Di-0-(n-hexadecyl)-2-0-,~~3-ethylaminopropyl)-glycerol Hydrochloride 'A. 1,3-Di-0-~n-hexadecyl)-2-0-~3-acetamidopropyl)-glycerol 1,3-Di-0-(~-hexadecyl)-2-0-(3-aminopropyl)-glycerol hydrochloride (1.0 g~, 1.6 mmoles) was added to a mixture of potassium ~arbonate (83~ mg., 6.0 mmoles) and benzene (75 ml.~.
~! Acetyl chloride ~150 mg., 1.9 mmoles) was then added a~d the resulting mixture stirred for one hour at reflux. Additional acetyl chloride ~150 mg., 1.9 moles) was added and the reaction mixture stirred for ano~her hour at reFlux. TLC analysis showed ' that the reaction was essentially complete. The reaction ' mix~ure was ~ooled, water (75 ml.) added~ and the resulting j.
mixture extracted with ether (3 X 100 ml.~. The combined ether extract was dried (MgSO4~, filtered and evaporated in vacuo to yieild the named compound i800 mg., 79~ yield; m.p. 53-54C., ir ~CHC13~ ¦
3400 and 1670 cm 1, n.m.r. (CDC13) ~ 1.97 (s, 3, -NHCOCH3)~. -; . I

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B. Title Compound 1,3-Di-0-~n-hexadecyl)-2-0-~3-acetamidopropyl)-glycerol (700 mg., 1.1 mmoles) was dissolved in ether (100 ml.) and tr2ated.
; with lithium aluminum hydrid~ (500 mg., 13 mmoles). Water . (100 ml.) was then added and the ~ixture extracted with ether ~2 X 100 ml.). The combined ether extract was dried (MgS04), .
. filtered, treated with hydrogen chloride gas and evaporated in ~acuo to a solid, which was recrystallizea from hot ethyl ... acetate [470 mg., 66% yield, m.p. 61-62C., n.m.r. (~DC13)~ 1.47 ~ (t, 3, -NHCH2CH3), elemental analysis calculated: 7~.51% C;
, 12.78% H; 2.11~ N; found: 72.47~ C; 12.56% H; 2.03% N~.
EX~MPLE 10 ;
: 1,3-Di-0-(n-hexadecyl)-2-0~
(3-isopro~vlamino~rovvl)-alvcerol Hvdrochloride ~ ~ , .
1,3-Di-0-~n-hexadecyl)-2-0-(3-aminopropyl)-glycerol :!! hydrochloride (700 mg., 1.1 mmoles) was dissolved in a solutisn .~ ~f acetic acid ~1.05:ml.), sodium acetate (350 mg., 4.3 mmoles) .
a~d acetone (1.3 ml.). Sodium borohydride ~1.25 g., 33 mmoles) was added in small portiQns until TLC a~alysis showed that all the 20 ~ 3-aminopropyl compound had been consumed. The reaction mixture was the~ treated with aqueous 2N sodium hydroxide (.20 ml.) and water (20 ml~), and extracted with e~her (3 X 40 ml.). The combined ether extract wa~ dried (MgS04), filtered, trea~ed with hydrogen chl~ride gas, and then evaporated in vacuo to a ~ solid, which was recrystallized from hot ethyl acetate 1210 mg., solid contained about 1/2 mole H20 per mole named pro uct, 28%
yield, m.p. 72-73C., n.m.r. (CDC13) ~ 1.42 (d, 6, -NHCH~CH3~2), elemental analysis calculated: 71.82~ C; 12.79% H; 2.04% ~;

found: 71.92% C; 12.46% H; 1.94% ~].

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,. i EXA*SPLE 11 1,2-Di-0-~n-hexadecyl)-3-0-(2-isopropylaminoethyl)-gl~cerol Hydrochloride ~j A. 1,2-Di-0-(n-hexadecyl)-3-0-all~l-glycerol

5 l~ Sodium hydride (1.78 g, of 50 wt. % dispersion in mineral j~ oil, 37 mmoles) was added at 60C. to a solution of 1,2-di-0-i~ (n-hexadecyl)-glycerol (10 g., 18.5 mmoles) in N,N-dimethyl-~i formamide (100 mll), and the resulting solution stirred for 20 ,j minutes at 60C. Allyl bromide (4.47 g., 37 mmoles) was then 't added dropwise and the resulting mixture stixred for 3 hours ., at 90C., cooled, cautiosly diluted with water (200 ml.) to quenchl ! the reaction, and extracted with ether (3 X 150 ml.). The combined ether extract was washed with saturated aqueous sodium chloride ~. solution, dried ~MgSO4), filtered and evaporated in vacuo to ~ an bil, which was purified by silica gel chromatography (elution 1. with benzene) ~10 g., 93% yield, oil, n.m.r. (CDC13)S 5.66-6.16 (m, 1, -OC~2CH=CH2), 5.25 ~d of doublets, 2, -OCH2CH=C~ ) and -~
. 4O03 ~a, 2~-OCH2CH=CH2~].
~ B~ 1,2-Di-0-(n-~exade~yl)-3-0-fv~ylme~yl-glycerol ~ Osmuum tetroxide (90 mg.,.354 mmoles) was added-to a solution of 1,2-di-0-(n-hexadecyl)-3-0-allyl-glycerol ~4~5 g., i 7.75 mmoles) in tetrahydro~uran:water (3:1, 120 ml.), and the ; resulting solu~ion stirred for 5~minutes at room temperature. I
~, Sodium periodate ~9 g.~. 42 mmoles) was then added and the reactiol -.' solution -~tirred for 16 hours at room temperature under nitrogen.
,. I
. The reaction solution was then diluted with water (150 ml.) and ex racted w~th ether (2 X 150 ml.). The combined ether ' extract was washed with water (150 ml~, dried (MgSO4) and i evaporated in ~acuo ~o an oil, which was purified by silica gel ~, chromatogxaphy (elution with ~enzene:ethyl acetate), [2.6 g., 57~ yield, waxy solid, ir (CHC13) 1735 cm 1, n.m.r. ~CDC13)~ 9.3B
,¦ (t, J = I Hz, 1, -OCH2CH0) and 4.07 (d, J = 1 Hæ, 2, -OCH~CHO)].
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C. Title Compound Sodium cyanoborohydride tO.l g., 1.6 mmoles) was added to , a solution of 1,2-di-0-(n-hexadecyl)-3-0-formylmethyl-glycerol , (1.5 g., 2.6 mmoles) and isopropylamine (0.89 g., 15 mmoles) , in methanol:tetrahydrofuran (1:1, 50 ml.), an~ the mixture stirred for two hours at room temperature. The pH was then adjusted , to 6 with 5N methanolic hydrochloric acid, additional sodium li cyanoborohydxide (0.1 g., 1.6 mmoles) added, and the re~ction ~ mixture ~hen stirred for another 60 hours at room temperature, !! filter~d, ~reated with aqueous 3N sodium hydroxide (10 ml.) and I saturated aqueous sodium chloride solutio~ (200 ml.~, and extracte~
¦ with Pther(2 X 150 ml.3. The combined ether extract was dried , (MgS04), fil~ered and evaporated in ~acuo to an oily solid, which j was purified by sil~ca gel chromatography (elution with ~enzene: ¦
i ethanol~ and dissolved in methanol. The ~olution was treated wi~n!
- , hydrogen chl~rid~ gas and evaporated in vacuo to yield a solid, 1~ , , which was recrystallized from ethyl acetate [400 mg., solid ~ contained about 1/4 mole H~O per mole named prod~ct, 23~ yield, i~ m.p. 71-72~., n.m.r. (CDC13) ~ 1.42 (d, 3 = 6 Hz, 6~ -NHCHrC~332), , elementaI analysis c~l~ulatad. 72.02~ C; 12.76% H; 2.1a~ N;
found: 11. 89~ C; 12~3~% H; 2.09~ Nl.
iEXAMPLE 12 i1,2-Di-O-(n-hexadecyl)-3-0-[2-(2-hydroxyethylamino)-jethyl]-glYcerol HYdrochloride ~In like manner to that described in Example 11 the named j compound was prepared by reacting 2-hydroxyethylamine with 1,2-di-O-(n-hexadecyl)-3-0-formylmethyl glycerol [solid contained about 1/2 mole H20 per mole named produc~, m.~. 125-126C., ~ ~lemental analysis calculated: 6g.54% C; 12.42% H, 2.07% N;
1 found: 6g.62~ C; 12.08% H; 2.29~ N~.
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1,3-Di-0-(n-hexadecyl)-2-0-(4-aminobutyl)-glyCerol Hydrochloride A. 1,3-Di-0-(n-hexadecyl)-2-0-~3-hydroxypropyl)-glycerol ~1 Borane methyl sulfide ~B~S) complex (6.5 ml., 68.5 mmoles) . was added at 0 to 5C. to a solution of 1,3-di-0-(n-hexadecyl)-,. 2-0-allyl-glycerol (10.82 g., 18.6 mmoles, prepared as in Example llA) in hexane ~190 ml.), and the resulting solution il stirred for 3 hours at room temperature. The reaction solution was then cooled again to 0 to 5C. and ethanol ~17.3 ml.) added l dropwis~ to decompose residual ~MS. The reaction solution wa-then treated with aqueous 3N sodium hydroxide (13 ml.) and 30 wt. ~ aqueous hydrogen peroxide (11 ml.), stirrea for 16 hours j, at reflux, cooled, and poured into ice water containing sodium ¦' bisulfite. The ice water solution was stirred until it gave ~ a negative starch-iodide test for peroxides, and then extracted with ether ~3 X 200 ml.). The combined ether extract was washed with water ~200 mlO), washed with sàturated aqueous sodium i! - I
chlo~ide solution ~200 ml.) r dried (MgSO4), filtered and ¦
I e~aporated in vacuo. The resulting pxoduct was purified by " silica gel chromatography ~elu~ion with benzene:ethanol) 15 g., 4S% yield, m p. 29~C~, n~m.r. ~CDC13)~3.80 ~tf J - 5 ~z, ~, ,' -OCH2.CH2CH2OH~ and 3.75 (t, 3 - 5 Hz, 2, -OCH2C~2CH~OH)].
. B. 1,3-Di-0-(n-hexadecyl)-2-0-E3-(p- os~loxy)propyl]-glycer i 1,3-Di-0-(n-hexadecyl)-2-0-(3-hydryoxypropyl)-glycerol ~5 ' (8.0 g., 13.4 mmoles) was added at 10C. to a solution of p-.toluenesul~onyl chloride (5.2~ g., 27.5 mmoles) and pyridine ~10 ml~
in methylene chloride (200 ml.), and the mixture stirred for 6Q
; hours at room temperature. Water ~200 ml.) was then added, the s, . . I

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methylene chloride and aqueous phases separated, and ~he latter ~, extracted with methylene chloride (2 X 150 ml.). The three methylene chloride layers were combined, washed with water ~2 X 150 ml.), dried ~MgSO4), filtered and evaporated in vacuo.
5 1 The resulting tosylate was purified by silica gel chromatography ,l (elution with benzene) 13.0 g., 30% yield, oil, ir (CHC13) 1130 ~¦ and 1350 cm 1, n.m.r. (CDC13) ,~ 7.53 (q, 4, protons on phenyl ,~ ring), 4.15 (t, 2t -503CH2CH~CH20-), 3.63 (t, 2, -~O3CH2CH2CH20-), 3.42 (m, 9, -OCHICH2OCH2Cl-H3lJ ), 2.45 ~s, 3, Ar-CH3), 1.90 ~ 2 il ~m, 2, -SO3CH2C~ C~20-) and 0.~0-l.S0 (~, 62, alipha~ic protons)].
. 1,3-Di-0-(n-hexadecyl)-2-0-(3-cyanopropyl)-glycerol ~ 3-Di 0-(n-hexadecyl)-2-0-,~3 (~-tosyloxy)propyl]-glycerol i' t3.0 g., 4.0 mmoles) was dissolved in a solution of sodi~m jg cyanide (0.5 g., 10 mmoles) in N,N-dimethylformamide (50 ml~), ~ and ~he resulting solu~ion stirred for 16 hours at 80C.~ cooled, diluted with wa~er (lOO ml.) and extracted with ether (3 X 100 ml.l.
The combined e~her extract was washèd consecutively with lN
hydrochloric acid (3 X 75 ml.) t saturated a~ueous sodium ~ ll bicarbonate solution (3 X 75 ml.), water t75 ml.) and saturated ' aqueous sodium chlsride solution (75 ml.), then dried (MgS04), ~iltered and evaporated in v~cuo to yield a waxy solid that j was used in ~he next step without further purification 12.0 g., I, ~3~ yield, ir -(C~C13~ 2250 cm 1~1~
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'I!j ~7 Ls~L3~8 ~1 sl sl D. Title Compound ' Lithium aluminum hydride ~800 mg., 21 mmoles) was added to a solution of 1,3-di-0-(n-hexadecyl)-2-0-~3-cyanopropyl)~glycerol ¦
~ t2.0 g., 3.3 mmoles) in ether ~100 ml.), and the mixture stirred ~ for 60 hours at room temperature. Enough water to quench thë
,~ reaction was added cautiously, followed by an additional 100 ml.
il of water. ~he resulting mixture was stirred for another hour at t room temperature and then extracted with ether (3 X 100 ml.~.
Il The combined ether extract was washed with saturated aqueous !', sodium chloride solution (3 X 75 ml.), ~ried (MgS04), filtered and evaporated in vacuo to an oil, which was purified by silica i gel chromatography (elution with benzene:ethanol) and then dissolved in ethanol. The solution was treated with hydrogen I chloride gas and then evaporated in vacuo to yield a solid, which lS ~I was recrystallized~frS~m ethyl acetate [444 mg., 21% yield, m.p.
1, 61.5_53,5C., n.m.r. tCDC13) ~ 3.67 (t~ 2~ -O~2CH2cH2~H2NH2~
'I 3.55 ~m, 9, -OCHlCH~OCH2C15H31]~), 3~10 (t, 2, -OCH2CH2C~2CH2N~2), ~ o-2.00 ~m, 4, -OCH2CH2CH2CH2~2~ ansl 0.80-1.5~ (m, 62, aliphati :
.l~ protons), elemental analysis calrulated: 72.23% C; 12.74~ H;
i, 2.16% N; found: 72~53~ C; 12.42% H; 2.1D% N~.
- ¦l ExaMPLE 14 '~I 1,2-Di-0-(n-hexadecyl)-3-0-1 (3-aminomethvl~enzYl)-qlYcerol Hydrochloride j A. 1,2-Di-0-(n-hexadecyl)-3-0-(3~cyanobenzyll-~lycers~l 'I Sodium hydride (1.056 g. of 50 wt~ ~ mineral oil dispersion, ¦
~ 22 mmoles) was added to a ~olution of 1,2-di-0-(n-hexadecyl)-'" ~lycerol (9.7~ g., 18 mmoles) in tetrahydrofuran (150 ml.) and the resulting solution stirred for 20 minutes at room ', temperature under nitrogen. m-Cyanobenzyl bromide (4.0 g., 20 '~ mm~les) was added and the reaction mixture stirred overnight at ,1 æ~

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;' room temperature under nitrogen. Water (200 ml.) was then added . cautiously and the resulting mixture extracted with ethyl acetate ~3 X 150 ml.). The comkined ethyl acetate ex~ract was , dried (MgSO4), filtered and evaporated in vacuo to an oil -ll (L2 g.), whlch was purified by silica gel chromatography i~ (elution wi~h benæene:hexane) {~.0 g., 68% yield, oil, ~¦ ir (CHC13) 2230 cm 1~, B. Title Compound - .
~ A solution o 1,2-di-0-(n-hexadecyl~-3-0-~3-cyanobenzyl)-i1 glycerol (l.0 g~ mmoles) in ether ~10 ml.) was slowly added ~I' under nitrogen to a suspension ~f lithium al~minum hydride ~, (0.057 g., 1.5 mmoles) in ether ~40 ml.), and the resulting .
i~ mixtur2 stirred for one hour at reflux under nitrogen and 1~ the~ cooled. ~ater (50 ml.) was added cautiously and the mix~ure !~ extracted with ethër (3 X 50 ml.). The combined etber extract ' was dried tMgS041, filtered and evaporated in va~uo to an oil, ~
~ which was purified by silica gel chromatography lelution with I
, ben~ene:ethanol) and then d1ssol~ed in ethyl acetate. The ~ ~olution was treated with hydrogen chloride gas and then ~ evaporated i~ vacuo to yield a solid, which was recrystallized ro~ ethyl acetate ~220 mg., 21% yield, m.p. 88-90C., elemental ' analysis calculated: 74.14% C; 11.87% H; 2.01~ N; folmd:
i 74.35~.C; 11.~4% H; 2.15% N]. .

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'' lS 7~.1411.87 2.01 73.89 11.43 1.99 ',',16 73.2011.85 1.9~ 73.17 11.53 2.28 7 72.2811.83 1.96 72.52 11.46 1~90 " 18 72.6311.48 2.17 72.62 11.81 2.43 !. 19 74.14 11.87 2.01 73.94 11.25, 2.02 . ~
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3L~ 35~ 1 . l , l EX~MPLE 27 ~ Di-0-(n-hexadecyl)-3-0- !
;; (4-aminomethylphenyl)-glycerol Hydrochloride ~ Al 1,2-Di-0-(n-hexadecyl)~3-0- (p-to5yl) -glycerol l, In like manner to that described in Example 13B the named .' compound was prepared by reacting 1,2-di-0-(n-hexadecyl)-glycerol i~ with ~-tol,uenesulfonyl chloride. Puxiication was accomplished ¦l by recrystallization from ethyl acetate ~m.p. 53-55C., ir l~ (C~C13) 1360 and 1180 cm 1~.
j B. 1,2~Di~0-(n-hexadecyl)-3-0-(4-cyanophenyl)-glycerol A mixture of 1,2-di-0-(n-hexadecyl)-3-0-~p-tosyl)-glycerol tl.4 g., 2.0 mmoles)~ sodium 4-cyanophenolate ~0.5 g., 3.5 mmole~) Il and xy1ene ~100 ml.) was stirred for 16 hours at r2flux. Since .i the reaction was not yet complete the xylene was removed by 15 il distillation and replaced by N,N-dimethylformamide (100 ml.), ~, and the resulting solution stirred for another 16 hours at 150C.¦
The reaction solution was then cooled, diluted with water (100 ml.) . and extracted with ether (2 X 100 ml~)~ The combined ether extract was washed consecutively with 3N hydrochloric aoid . (100 ml.), 10 wt~ % aqueous sodium bicar~onate solution ~100 ml.) ',1 and water ~100 ml.), dried (MgS04), filtered and evaporated i i~ vacuo to an oil, which was purified b~ silica gel ch~matography ~, telution with benzene~ ~0.65 g., 50% ~ield, m.p. 53-55C., ir Il (C~C133 2210 cm 1].
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l~ C. Title Compound il 1,2-Di-O-(n-hexadecyl)-3-0-~4-cyanophenol)-glycerol (0.60 g.~ 0.93 mmole~ was added to a suspension of lithium ~1 aluminum hydride (O.3 g., 7.9 mmoles) in ether (25 ml.), and !~ the resulting mixture stirred for 30 minutes at room temperature;
il Water ~25 ml.) was then added cautiously, the ether and aqueous phase$ separa~ed, and the latter extracted with ether (3 X 25 ml.
i! and ethyl aceta~e ~25 ml.). The fiYe organic extracts were i~ combined, dried (MgS04), filtered a~d evaporated in vacuo to an I~l oil, which waC~ dissolved in ether. The solution was ~reated ' wi~h hydxogen chloride gas, causing precipitation of a solid ,~-tO~41 g., 64% yield, m.p. 110-112C., n.m.r. ~CDC13) ~ 4.02 ! (s 2, -CH2NH2~, elemental analysis calculated- 73.91% C;
,i 11.81~ H; 2.05~ N, found: 73~6~% C; 11.71% ~; 2.14% N3.
15 l~ EX~MPLES 28-30 `
, In like manner to that described in Example 27 B-C the following compounds were pxepared from the appropriate tosylate (prepared as in Ex~mple 27A) and sodium cy~nophenolate~

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. , ~' X~PLE 31 '1,2-Di~0-(n-hexadecyl)-3-0-[4-(3-aminopropyl)phenyl]-glycerol Hydrochloride ,' In like manner to tha-t described in Example 27 the named . compound was prepared by using sodium 4-(2-cyanoethyl)phenoiate in place of sodium 4-phenolate (m.p. 153-155C., elemental analysis ralculated: 74.37~ C; 11.91~ H; 1.97% N; found:
¦l 74.13% C; 11.44% H; 2.08% N1.
Ii EXANPLB 32 j . ~ 1,2-Di(n-hexadeoyloxy)-3--(3-aminomethylbenzylamino)-~ropane Dihydrochloride : 3 1,~-Di-0-(n-hexadecyl)-3-0-~-tosyl)-glycerol (3.48 g., 5.0 mmoles~ was added to a solution o~ m-xylylenediamine (0.68 g.
I' 5.0 mmoles) in N,N-dimethylormamide ~20 ml.). The resulting I mixture was stirred for one hour at gO~C. and then poured in~o ice water (150 ml~ ausing the formation of solids which were isolated by filtration, purified b~ sili~a gel chromatography ~elution with ben2ene:ethanol) ar~d ~hen dissolved in ethyl l¦ aceta~e. The solution was treated with hydrogen chloride. gas ,~ and then evaporated in ~acuo to yield a ~olid, which was re-?3!
i. crystallized from ~thyl acetate l{0~29 g., 8% yield, m.p. 78-80C.

i, n.m.r. (CDC13) ,~ 4.24 ~s, 2, Ar-~H2N'~-) and 4-37 (s, ~, ', Ar-CH2NH2),.elemental analysis ~alculated: 70,55% C;

~ 57% ~; 3.83% N; ~ound: 70.64~ C; 11~9~ H, 3.62% N3.

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I' I . EX~5P~E 33 ~. 1,2-~i-0-(n-hexadecyl)-3-0-(3-isopropylamino-l! 2-hydrox ~ropyl)-~lycerol H~drochloride ,~ . ' i ! A. 1,2-Di-O-(n-hexadecyl)-3-0-~2,3-epoxypropyl)-~lycerol l, A solutlon of 1,2-di-0-(n-hexadecyl)-3-allyl-glycerol i~ (5.8 g., 10~0 ~moles) and m-chloroperbenzoic acid (1.86 g., 10.8 m~,oles) in benzene (50 ml.) was stirred at reflux for 16 ~, hours The reaction mixture was then cooled, treated with - il satura~ed a~ueous sodium bisulfite solution tlO ml.) and '! ~aturated aqueous sodium bicarbonate solution (50 ml.), and , extrac~ed with ether (3 X iO ml.). The combined-ether extract !~ was washed with water ~100 ml.), washed with saturated aqueous ~odium chloride solution (100 ml.), dried (MgS04,), filtered and `~ evaporated in vacuo to an oil t4.9 g., 82~ yield, olefinic ¦J, protons absent by n.m.r. analysis), which was puri~ied by silica ! gel chromatography (elutiQn with ~enzene:ethyl acetate) ~4.2 g., 70% yie d, oil-solidified on standing). -,~ Bo Title Compound .. A solution of 1,2-di~O-(n-hexadecyl)-3~0-(2,3-epoxypropyl)-,. glycerol ~2.0 g., 3.35 mmoles) i~l isopropy~amine (40 ml.) was : ~' heated in a stainless ~teel bomb for 16 hours at lOO~C., cooled, i~ concentrated in vacuo and dissolved in ether (100 ml.). The ¦. e~her solution was washed wi~h.lN hydrochloric acid ~100 ml.), '' dried (MgS04), filtered, treated with charcoal, filtered again, l~ and then c~led by immersion of the 1ask in a Dry Ic~-acetone ~1 bath, causing-for~,ation of a pr~cipitake. The precipitate was isolated ~y ~iltration ~1.3 g.) and purified by silica gel chromatography (elution with benzene:ethanol) 1720 mg., solid l contained abou 1/2 mole H20 per mole named product, 31~ yield, ~ m.p. 55-57~C~, n~m.r. (CDC13)~ 1.45 ~d, 6, -NHCH~CH312), elementai ,l analysis calculated: 70019% C;.12.50% H, 2.00% N: found: 70.10 ,I C; 12.19% H; 1.87~ ~].
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3~8 , In like manner to that described in Example 33B the following compounds were Prepared by reacting the appropriate '. 2,3-epoxide (prepared as in Example 33A) and alkylamine:

5 ~' . n-hexadecyl-0-CH2 OH
,¦ . CH-0-CH2~HC~2NHR3 il ~ n-hexadecyl-O-CH2 ii ' ' CH2-~)-CH2~HCEI2NHR3 10 ~ n-hexadecyl-0-1H II

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1,2-Di-O~(n-hexadecyl)-3-0-(3-amino-2-hydroxypropyl)-gly~ol Hydrochloride A. 1~2-Di-O-(n-hexadecyl)-3-0-(3-azido-2-hydroxYpropyl) ~lycerol A solution of sodium azide (0.5 g., 7.7 mmoles) in water (5 ml.) was added to a refluxing solution of 1,2-di-0-(n-hexadecyl)-3-0-(2,3-epoxypropyl)-glycerol (3.3 g., 5.5 mmoles) in 1,4-dioxane (100 ml.), and the resulting solution stirred at reflux for 16 hours. Since the reaction was not yet complete, additional sodium azide (O.5 g., 7.7 mmoles) was added and the reaction stirred at reflux for another 16 hours. The rèaction solution was then cooled, concentrated in vacuo, diluted with . water (100 ml.) and extracted with ether (3 X 100 ml.). The combined ether extract was washed with water (100 ml.), dried IMgSO4), filtered and evaporated in vacuo to an oil which -1 solidified on standing [2.2 g., 62% yield, ir (CHC13) 2105 cm ].
B. Title Compound Lithium aluminum hydride (300 mg., 7.9 mmoles) was added to a solution of 1,2-di-0-(n-hexadecyl)-3-0-(3-azido 2-hydroxy-propyl)-glycerol (2.2 9~, 3.4 mmole ) in ether (100 ml.), and the resulting mixture stirred for one hour at room temperature.
Ethanol (5 ml.) and water (200 ml.~ were added to guench the reaction, and the mixture then extracted with ether ~2 X 100 ml.) .
The combined ether extract was dried (MgSO4), filtered and evaporated in vacuo. The resulting product was purified by silica gel chromatography ~elution with benzene:ethanol) and then converted to the hydrochloride salt [800 mg., solid ~ 3~8 contained about 2 moles H20 per mole named product, 34% yield, m.p. 149-150C., n.m.r. (CDC13) 4.00-4.35 (m, 1, ¦ -OCH2CHOCHCH2NH2), 3.33~3-73 (m~ 11~ ClsH31CH2CH2CH[CH2C15H31 ¦ CH2OCH2-), 3.03-3.25 (m, 2, -OCH2CHOHCH2NH2) and 0.87-1.67 ¦ (m, 62, aliphatic protons), elemental analysis calculated:
¦ 66.48% C; 12.33~ H; 2.04% N; found: 66.68% C; 11.85% H; 2.02% N] .
¦ EXAMPLE 39 1 3-Di-0-(n-hexadec 1)-2-0-(3-amino-2-h drox ro 1)~ 1 cerol I , Y , , Y y~ Py g Y
¦ In like manner to that described in Example 38 the named ¦ compound was prepared-from 1,3-di-0-(n-hexadecyl)-2-0-(2,3-¦ epoxypropyl)-glycerol (prepared as in Example 33A) (free base, ¦ m.p. 61-63C., elemental analysis calculated: 74.33% C; 12.96%
¦ H; 2.28% N; found: 74.49% C; 13.10% H; 2.12% N).
¦ - EX~MPLE 40 ¦ 1,2-Di-0-(n-hexadecyl)-3-0-(2-amin~æropyl)-glycerol Hydrochlorlde A. 1,2-Di-0-(n-hexadec~1)-3 0-~2-(p-tosyloxY)propyl]-glycerol In like manner to that described in Example 13A and B, 1,2-di 0-(n-hexadecyl)-3-0-allyl-glycerol was reacted with BMS, and the resulting 2-hydroxypropyl and 3-hydroxypropyl compounds converted to their corresponding tosylates. A separation was not attempted at this stage; the mixture of tosylates was used directly in the next step.

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B~ 1,2-Di-O~(n-hexadecyl)-3-0-(2-azidopropyl)-glycerol ~he resulting mixture of tosylates (3.0 ~., 4.0 mmoles) was dissolved in N,N-dimethylacetamide (50 ml.) and treated with a solution of sodium azide tO.326 g., 5.0 mmoles) in water (5 ml.)l 5 ; for 16 hours at ~0C. The reaction solution was then cooled, .¦
i diluted with water (200 ml.), and ex~racted with ether (2x150 ml.) il ~he ~ombined ether extract was washed with water, dried (MgS04), .~ filtered, and evaporated in vacuo to an oil [2 g., 81~ yield;
i~ ir (CHC13~ 2100 cm 13, a mixture of the 2-azidopropyl ana 3- .
, azidopropyl compo~nds, which was used without further purificatio . in the next step~ - .
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, The resulting mixture of azides ~2 g., 3.2 mmoles~ was dissolved in ether ~100 ml.), treated with lithium aluminum , hydride (0.4 g., 10~5 mmoles~, and allowed to sti~-for 2 hours at room temperature. Excess hydride was destroyed by cautious addition of ethanol ~10 ml.) and water (150 ml.), and the mixture I
then extracted with ether (2xlO0 ml.). The combined ethex extxactl ~ was dried ~MgSO43, filtered~ and concentrated in ~a~uo ts an oil ,, (1.8 g.), which was purified by silica gel chromatography ~elutior ,' with benzene:ethanol) and then converted to the hydrochloride salt I
by dissolution and treatment with hydrogen chloride gas. The ' salt was recrystallized rom ethyl acetate (0.21 g., solid containec '., about 1~2 mo}e H20 per mole named product, 10% yield, m.p. 56-58~C., elemental analysis calculated: 71.03% C; 12~70% H; 2.18~ N;
~ou~a: 71.11~ C; 12.91~ ~; 2D1~% N).

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EXAMP.~E 41 .
, 1,2-Di-O-(n-octadecyl)-3-0-(2-aminopropyl)-~lycerol Hydrochloride : In like manner to that described in Example 40A>1,2-di-0-. ~ ~n-octadecyl)-3-0-~2-hydroxypropyl)-glycerol was prepared from ~. 1,2-di 0-~n-octadecyl)-3-0-ally~-glycerol. The named compound 1i was prepared from 1,2-di-0-(n-octadecyl)-3-0-(2-hydroxyp~opyl)-¦~ glycerol in like manner to that described in Example 40 B-C ~solid Ij con~ained about 1 mole H20 per mole named product, m~p. 65-67C. t j. elemental analysis calculated: 71.1g% C; 12.80~ H; 1.98% N;
1~ found: 71.12~ C; 12.52~ H; 1.92% N). . .
¦j EXAMPLE 42 1,2-Di(n-hexadecyloxy)-3~a~inopropane Hydrochloride ¦i In like manner to that described in Example 4OB, 1,2-di-0- .
l~ ~n-hexadec~ 3-0~ osyl)-glycerol was converted to 1,2-di-'i (n-hexadecylQxy)-3-azidopropane. This inter~ediate was converted il to the title compound in li~e manner to that described in Example i 40C (m.p. 78-80C., elemental analysis calculated: 72.93% C; .
12.94% ~; 2.43% N; ~oun~ 73.0a~ C; 13.08% H; 2.65~ N). .

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1,3-Di~n-hexadecyloxy)-2-aminopropane Hydrochloride In like manner to that described in Example 42 the named compound was prepared from 1,3-di-0-(n-hexadecyl)-2-0-t~-tosyl)-glycerol (prepared as in Example 27A) (m.p.S8-60C., elemental analysis calculated: 72.93% C; 12.94% H; 2.43~ N; found:
72.65% C; 13.02% H; 2.59% N).

1,2-Ditn~hexadecyloxy_-4-aminobutane Hydrochloride In like manner to that described in Example 42 the named compound was prepared by using sodium cyanide in place of sodium azide (m.p. 86-87C., elemental analysis calculated: 73.25% C;
12.97% H; 2.37% N; fou~d: 73~52~ C; 12~64% H; 2.50% N).
EX~MPLE 45 1,3-Di(n-hexadecyloxy)-2-(3-a _ nopropvlamino)propane Dihydrochloride A. 1,3-Di(n-hexadecyloxy)-2-(2-cyanoethyla _n )propane l A mixture of 1~3-di(n-hexadecyloxy~-2-aminopropane (500 mg., ¦ 0.93 mmoles), acrylonitrile (75 ml.) and 2 wt. % aqueous sodium ¦ hydroxide solution (75 ml.~ was heated to 60C. Tetrabutyl ¦ ammonium hydroxide (1 ml. of 40 wt. ~ aqueous solution~ was then ¦ added and the resulting mixture stlrred for 15 minutes at 90C.
¦ The reaction mixture was then cooled, causing precipitation of ¦ solids, which were isolated by filtration and found (TLC) to ¦ contain a large quantity of unreacted starting material. Using 1 fresh acrylonitrile and aqueous sodium hydroxide solution in eac~
¦ cycle, the solids were treated two more times by the above ¦ procedure.

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The third cycle solid product was purified by silica gel -chromatography (elu~ion with toluene:eth~l acetate) [200 mg., 36% yield, m.p. 45-46C., ir (CHC13) 2250 cm 1, n.m.r. (CDC13~ S
,~ 3.07 (t, 2, -NHCH2CH2CN) and 2.53 (t, 2, -NHC~2C~2CN)].
B. Title Compound ¦
A mixture of 1,3-di(n-hexadecyloxy)-2-(2-cyanoethylamino)-propane (200 mg., 0.34 mmoles), tetrahydrouran (10 ml.), ethanol (20 mi.) and Raney nickel catalyst (0.2 g.~ was saturated with ,' ammonia gas and then hydrogenated (50 psi) for about 4 hours at room temperature. The reaction mixture was then filtered and evaporated in vacuo to an oil, which was purified by si~ica gel chromatography (elution with toluene:ethyl acetate: ethanol:
methanol) and then dissolved in ethyl aceta~e. The solution was , treated with hydrogen chloride gas, causing precipitation of l solids ~10 mg. r solid contained about 2.5 moles H20 per mole named product, 4% yield, m.p. 235~236C,, elemental analysis ~alculated: 63.65% C; 12.51~ H; 3.90~ N; found: 63.60~ C;
' 1~.84~ ~; 3.75~ ~].
j! . EXAMPLE 46 , t .
1,2-Di-O-(n-hexadecyl~3-0-~4-amidinophe~yl)-~lycerol Hydxochlori~e ~, A ~olution of 1,2 di-0-(n-hexadecyl3~3-0-(4-cyanophenyl)-glycerol (3.5 g., 5.45 mmoles~, ethanol S lo ml . ) and 1,4-dioxane tlOO ml.) w~s saturated with hydrogen chloride gas at ~ 0C., and allowed to react ~or 16 hours at mbient ~emperature.
The reaction solution was then e~aporated in vacuo to an oil, thei oil dissolved in ethanol (100 ml.), and the resulting solution ' saturated with ammonia gas, stirred for 3 hours at reflux, diluted ., ~ ~ D
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il l ! with water (150 ml.), evaporated in vacuo to remove the majority , of the ethanol, and extracted with chloroform t3 X 150 ml.3. The `' combined chloroform extract was dried (MgS04), ~iltered and evaporated in vacuo to yield a solid, which was purified by , silica gel chromatography ~elution with benzene:ethanol) and i; then dissolved in ethyl acetate. The solution was treated with hydrogen chloride gas and then evaporated in vacuo to yield a ~, solid; which was recrystallized from ethyl acetate [1.0 g., 26%
!~ yield, m.p. 220-2~2C., ir ~CHC13) 1670 cm 1, elemental analysis ~, calculated: 72.53~ C; 11.45~ H; 4.03~ N; found: 72067% C;
, 11.38% H; 4.12~ N].
EXAMPLE 47 . .
. 1,2-Di-0-(n-hexadecyl)-3-0- !
'; ~3-amidinobenzyl?-glycerol Hydrochloride .
, .The named compound was prepared ~rom 1,2-di-0-~n-hexadecyl)-1 3-0-(3-cyanobenzyl)-glycerol in like manner to that described in Example 46 tsolid contained about 2 moles H20 per mole ~amed product, 20% yield, m.pO 155-157C., ir (C~C13) 1670 cm 1, . elemental analysis calculated: 69.27% C; 11.48% X; 3.76% N;
~0~, found: 69.11~ C; 10.63% N; 3.B3% N3~
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, EXA~IPLE 48 1,2-Di-0-(n-hexadecyl)-3-0-[3-(1-hydroxy-2-t-butylaminoethyl)-benzyl]-glycerol Hydrochloride ., . I
i A. 1,2-Di-0-(n-hexadecvl)-3-0-(3-formYlbenzvl)-alvcerol A solution of 1,2-di-0-(n-hexadecyl)-3 0-(3-cyanobenzyl)-glycerol (5.0 g., 7~6 mmoles) and diisobutylaluminum hydride ~I , .
i; (1.17 g., 8.2 mmoles) in benzene (25 ml.? was stirred for 16 ~ hours at ambient temperature. The reaction mixture was treated - i~ with methanol (4.22 ml.) and water (2.5 ml.~ and stirred to I0 ll decompose unreacted hydride, and then filtered and extracted .

!', with benzene (3x25 ml.). The combined benzene extract was dried I` . .
(Na2SO~ iltered and evaporated in vacuo to an oil, which was j' purified by silica gel chromatography (elution with benzene) i [2~0 g., 40% yield, oil, ir (CHCl3~ 1700 cm , n.m.r. (CDCl3) lS ', lO.l (s, l, -ArCH )~. .

~, B. 1,2-Di~0-tn-hexadecyl)-3-0-~3-(1,2-epoxyethyl)-l~ . benz~l]-glycerol ~i A s~spension of sodium hydridë~-(3.23 g~ o~ a 57 wt. % disper-¦
j sion in mineral oil, 67 mmoles) in dimethylsulfoxide (117 ml.) was' ~0 '. heated ~nder a nitrogen atmosphere at 70 to 75C. until hydrogen I --l', evolution stopped (45 min.). ~etrahydxofuran (88 ml.) was added ; and the mixture cooled to 0 to 5C. Trimethylsulfonium iodide ' ~13.67 g., 67 mmoles) was then.added in portions~ ~ollowed by ' rapid addition of a solution of 1,2-di 0-(n-hexadecylj-3-0-(3-25 ~ fo~mylbenzyl)-glycerol ~7.0 g., 10.6 mmoles) in tetrahydrouran (58 ml.); The resul ing mixture was stirred for 16 hours at !; room temperature, poured into water (200 ml.) and extracted with ether ~3x180 ml.~. The combinea ether extract was washed . with water (2xlO0 ml~ and saturatéd aqueous sodium chloride l! ~
'' solution (lO0 ml,), dried (MgS0~), filtered and evaporated in vacuo to ~n oil (7.0 g., 98% yield), which was suiciently pure to be used in..the next step~

I' C Title Compound "
A mixture of t-butylamine (30 ml.) and 1,2-di-0-(n-hexadecyl), 3 0-~3-~1,2-epoxyethyl)-ben~yl]-glycerol (2.0 g., 3~0 mmoles) 1, was heated for 9 hours at 100C. in a steel bo~b. The reaction S ~' mixture was cooled, t-butylamine removed by evaporation in vacuo, ¦

and the resulting oil purified by silica gel chromatography (elution with benzene:ethanol) and then dissolved. The solution ~' was saturated with hydrogen chloride gas and then e~aporated ,¦ in vacuo to yield a solid, which was recrystallized from ethyl ¦ acetate 1630 mg., solid contained about 1 mole H20 per mole named ~, product, 27~ yield, m~p. 49-51C.,-n.m.r. (CDC13) ~ 1.47 ~~ ~s, 9, -C~C~3]3), ele~ental analysis calculated 71.99% C;

! 11. 83% H; 1.75% N; found: 71.86% C; 11.30% ~; 1.69% Nl.

~ EXAMPLE 49 j3 1,3-Di-0-(n-hexadecyl)-~-0-~3-tl-hydroxy-2-t-butylaminoethyl)-benæyl]-glycero ~ dxochloride .
In like manner to that descri~ed in Example 48 A-B, 1,3-di-'1 0~ hexadecyl)-2-0-(3-cyanobenzyl)-glycerol ~pr~parea as in ~ Example 14A) was converted to 1,3-d~-U-~n-hexadecyl)-2-0-[3-(1,2-' epoxyethyl)-benzyl]-glycerol. The title compound was prepared by reacting said epoxy compound with t-butylamine in like manner to that des~ribed in Example 48C (solid contained about 1 mole i; E~0 per mole named product, m.p. 43-45C., elemental analysis l~ calculated: 71.~9~ C; 11.83~ H; 1.75% N; found: 72.06~ C;
, 11.43% H; 1.71% N).
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; In li~e manner to that described in Example 48C the named . compound was prepared by using isopropylamine in place of t-butylamine (solid contained about 3/4 mole H20 per mole named ' product, m.p. 53-559C., elemental analysis calculated: 7~.17%
, C; 11.19% H; 1.79% N; found~ 72.11% C; 11.55% H; 1.92% N).
j! EXAMP~E 51 . I .
~ 2,3-Di(n-hexadecyloxy)propyl3-4-aminomethyl-4-li A~ ~-[2,3-Di(n-hexadecyloxy)pro~ cano-4-phenylpiperidi -.,e ! A mixture of 1,2-di-0-(n-hexadecyl)-3~0-( -tos~l)-glycerol (6.96 g., 10 mmoles), 4-cyano-4-phenylpipexidine hydrochloride ., (2.23 g., 10 mmoles), triethylamine ~2 ml.) and N,N-dimethyl-.i formamide ~40 ml.) was stirred for 16 hours at g5 to ~OO~C.
j~ The reaction mixture was then cooled, diluted with water (200 ml.) and ex~racted with ethyl acetate (3x150 ml.). The comkined ethyl ¦
. acetate extract was dried (MgSO4), fil~ered and evaporated l~ in vacuo to an oil ~6.g.), which was purified ~y column chroma- ¦
tography (elution with benzene:ethyl acet,~te~ [oil, ir (CHC13) ,~, 22~0 ~m~~
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In like manner to that described in Example 51 the following com-pounds were prepared from the appropriate 1,2-di-0-~n-alkyl or alkenyl)-3-0-~p-tosyl)-glycerol ~prepared as in Example 27A):

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formylmethyl-glycerol with di(2-hydroxyethyl)amine (solid . contained about l/4 mole H20 per mole named product, m.p.
194 195C., elemental analysis calculated: 69.05~ C; 12.20% H;
1,l 1.96% ~; found: 69.12% C; 11.76% H; 1.~0% N). .
Ij ~ EXAMPLE 57 In Viv~ Activity of 1,3-Di-0-(n~hexadecyl)-2-0-l~ (3-amino~ro~vl)-alvcerol Hvdrochloride Aqainst EMC Virus ' Formulation as an emulsion was accomplished by mel~ing !! and mixing equal part~ of the named compound, polysorbatè 80, 1'. and glycerin, and then dispersing the mix~ure in hot wa~er under -vigorous mixing. The formulation was then ad~usted to final i.
,j concentrations of 0.14M sodium chloxide and 0.OlM sodium .
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i, phosphate, pH 7. Further dilutions were made with 0.14M sodium ~, chloride -O.OlM sodium phosphate, pH 7 buffer solution.
.
~, Three groups of ten female albino mice ~20-25 g. body .
~' weight) were gLven 0.5 ml~. intraperitoneal injections containins ; dosage levels of 1.5, 5 and 15 mg. of the named compound~kg.
body weight, respectively. A fourkh control group of ten 'i mice was given no such injection. ~igh~een to ~wen~y-four ~, houxs later all four groups were challenged wi~h 0.2 ml.
~u~cutaneous injection containing 20 times the LD50/ the dosage ., 7evel causing a 50% death rate i~ unprotected mice in t~ days, ' of encephalomyocarditis (EMC) virus. Survival data were recordea .
over the next~ten days and the relative survival (Sr) calculated:
Dosage Level of :;Named Compound Sr ~a~erage of seven experiments) . .
.lS mg.~kg~ 61 . ~ 45 1.5 2 p 3~8 . . . , i - !
;, Antiviral activity is expressed as the relative survival . (Srl in experimental groups compared to the control on the ~, tenth day after challenge, Sr is defined by the formula '' Sr = r Sx ~ = o 10 i=l to 10 ~ X 100 j, . 100 ~ 100 -~ei _ , i=l to 10 . wherein Sr = relative survival i Sx = percent survival a~ter ten days in experimenta 1 lo I! group ~! - Xi = number of survivors on the ith day in ¦¦ experimental group ,' ei = number of survivors on the ith ~ay in control , t~ group ~; EXAMPLES ~8-86 .
~i In like manner to that described in Example 57 the in vivo il activity against EMC virus was determined for the compounds listed below.
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~,f~ 35f~3 .! EXAMPLE 87 Reduction of Virus Yield on Human Polyp Cells In Vitro by 1,3-' Di-0-(n hexadecyl)-2~0-(3~aminopropyl)-glycerol Hydrochloride , Growth medium was prepared by supplementing Eagle~s minimum S essential medium ~100 ml.) with lOOX concentrated antibiotic antimycotic solution ~2 ml.), 200 mM glutamine solution (1 ml.), lOOX concentra~ed nonessential amin~ acids solution ll ml.), " 100 mM sodium pyruvate solution ~1 ml.), and heat-inactivated .
;,fetal calf serum (10~). Each well of 96-well microtiter i¦plates was seeded with about 50~000 hum~n nasal polyp ,cells suspended in 0.2 ml. growth medium. The plates were then Ij incubated for 8-}0 days at 37~C. in a 5% CO2 atmosphere to 'establish monolayers of cells.
At the end of the 8-10 day cell growth period confluent lS l~monolayers on the plates were washed ~our times with phosphate ! buffered saline and immediately afterward treated with 0~2 ml.
i~per well of mainte~ànce medium ~ontaining 10, 5.0, 1~0, 0.5, t 0~1 and O~g./ml. of the named compound, respectively.
~'iThe maintenance medium ~as ide~tical to.the growth medium des- .
'"cribed above except that the level o~ said fetal calf serum was ~2%. The plates were incubated for another 18 hours at 37~.~
, and the monvlayers then wash~d four times with phosphate buffered j saline to remove the named compound~ challenged with a compositionl 'Icontaining about 1000 times the TCID50, i.e. the dosage level ! causing a 50% infection rate in unprotected cultures, of .vesicular-stomatitis virus (VSV) for a two hour (37~C.) adsorption.
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¦ plates of L-929 mouse fi~roblasts. The L-929 mouse cultures ¦, were scored microscopically and analyzed about three to four j days later, with the f~llowin~ percentage decreases în virus . ¦ yield (with respect to the control) determined for the five ~ conc~n~rations.o named compound tested:
, P~rcentase Reduction of ~irus Yield ncentra~~lo~7~ ~und ,5 lo 5.0 1.0 O.5 0.1 Ii !i 94% g~% 84% 75% e68 ji In lik4 mannër to th~t described i~ Example 87 the reduction ¦ of virus yield on human polyp cells in vitro was d~termined for i the compounds listed belsw.
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ol Z Z I Z~ Z Z Z ~ Z Z

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G

!l EXAMPLE 122 .

!` Ability of 1,3-Di-0-(n-hexadecyl)-2-Q-(3-aminopropyl)-gly~erol . _ Hydrochloride t.o Induce Circulatin~ Interferon. _ i ' A mixture of equal weights of the named compound, polysorbate 5 80 and glycerol was fused and then homogenized in hot 0.14~5 sodium chloride containing O.OlM sodium phosphate, pH 7 (PBS).

.The re~ulting oil-in-water emulsi~n w~s readily diluted with PBS
Il for aaministration.
j, Female Swiss mice ~20-25 g. body weight) were injected .
'l~ 5 ml~, intraperitoneal) with ~. quantity o the above diluted emulsion containing 25 mg. of the ~amed compoundfkg. body weight.
¦iEigh~c twelve, sixteen and twenty hours a~ter injection samples of plasma were withdrawn from four mice and po.oled. Serial , dilutions in ~-15 (Leibovitz) medium containing 5% ~etal calf 1~ serum were incubated in microtiter plates over~ight at 37C.
~on confluent m~n~làyers of L-929 mouse fibroblasts. The mono-layers were then washed with protein-free medium~ challenged ¦ with 10 times the TCID50, i.e. the dosaye level causing a 50%.
..infection rate in .unprotected cultures, of vesicular s~omatitis virus (VSV) for a one hour (37C.~ adsorption period, washed, .retrea~ed with L-15 medium-containing 5~ fetal calf serum and the~ incubated again f~r 48 hours at 37C. The L-929 culturas were then scored microscopically for ~iral cy~opathology and .analyzed, with the plasma inter~eron level, the reciprocal of the I
- .plasma dilution conferring 50% protec~ion to the L-g29 monolayers,¦

. determined A second experiment followed the above procedur`e, except ;~hat the mice were injected with 10 mg. of the name~ compound/kg.
body weight and samples of peritoneal wash we~e taken from four j.
,mice and pooled at six, nine, twelve, fifteen and eighteen hours ,1 ~

~ 3 ~ t li l . .

after i~jection. The samples were taken by exposing the ,iperitoneal membrane, in~ecting 1 ml. of Hank's balanced salt solu~ion containing 100 penicillin units~ml. and 100 ~g. ¦ -streptomycinJml. into the ~eritvneal cavity, briefly massaging .~the abdomen, and then aspirating ~he peritoneal w~sh.
li The following data were obtained from these two experiments:
i Inter~eron Levels (units~ml.) ; Source of Time (hrs.) after Injection . .'Interferon 6 8 ~ ~ 15 16 13 2~ .
! Plasma - 34- 67 - 52 - 40 I'Peritoneal i wash ~16 -768 320 448 - 448 ' ~3 ~a~
'. ~n like manner to that described in Example ~22 the ability , to induce eircula~ing interferon was determined for the comp~unds ii ...
ilisted below.

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Il~
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i~ ~ ''J ~J V `~ rt , ~1 ~ _ ~ol ~ QD ~ ~ ~ ~D r~

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o . $~o~ i i,, - ~ ~ .
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1. ~

~2~3~ 1 , , 1, .EXAMPLE 130 Enhancement of Poly.inosinic-Polycytidylic Acid ~Poly (I:C)J- l induced Cellular Resistance to Viral Infection by 1,3-Di-0- 1 . (n-hexadecyl)-2-0-(3-aminoprop~ glycerol Hydrochloride Ij Growth medium was prepared by supplementing Eagle's minimum ~, essential medium (100 ml.) with lOOX concentrated antibiotic-. antimycotic solution (2 ml.), 200 mM glutamine solutLon (1 ml.),ald ,~heat-inactivated fetal calf serum (5%)O Mouse L-929 fibroblasts . ~I were suspended in yrowth medium, and each well of 96-well microtiter ¦'plates was seeded with 0.2 ml. of said suspension containing ~0,000 ',to 30,000 cells. The plat s were incubated for 2 to 4 days at 37Ç.
' in a 5% C02 atmosphere to establish monolayers of cells. The plates were washed four times with phosphate buf~ered saline ' immediately prior to treatment~ . .
'. Poly (I:C) was prepared at concentrations o~ 5.0, 1.0, 0.2 and 0.04~ug.Jml. in the medium described above minus calf serum. I
i, O.1 ml. of each dilution was combined in a checkerboard arrange- ¦
ment on the L-92g cell monolayers wl;th 0 o l ml~ dilutions containing . 20.0~ 4.0, 0.8, 0~16 and 0.032~ug. o~ the named comp~und per ml.
-said serum-free medium~ Control wells were exposed to either poly~
~ .
' (I C) or the named compound alone. ~he plates were incubated ' fox 6 hours at 37C. in a 5~ CO2 atmosphere, washed foux times ' with phosphate buffered saline, and refed with 0.1 ml. per.weil ,' growth medium containing 2% fetal calf serum. After 18 more hours of incubation, ~he plates were scored for toxicity and then challenged with 0.1 ml. per well ~f a vesicular stomatitis ~irus (VSV) suspension containing 10 to 30 times the TCID50 ~tissue culture infective dose causing a 50% infection rate). The ., , . . I

, . . .

' plates were incubated ~or another 3 to 4 days and then scored microscopically for cytopathogenic e~ect (CPE). Cells protected i ' from virus in~ection were free of CPE. The minimum protective dose (MPD) of poly (I:C) alone was noted, and the amount of . enhanced or augmented antiviral acti~ity caused by combination ,;with the named.compound recorded for each dllution leYel of said ~ named compound.

~, .
!~ Enhancement of PO1Y ~ C~-indUCed ~,, Cellular Resistance to Viral Infection '` Concentration ( ~ ~./ml.) of Named Comnound f ~
~'2~.0 4.0 0.~ 0.16 ~.032 ,125X 125X 12~X 5X. C5X .
Note: combining poly(I:C) with named compound provides same . antiviral effect as increasing poly tI:C) concentratio~ indicated .
number of times. ..

In li~e manner to that described in Example 130 the enhance-'ment of poly~I:C)-induced cellular resistance ~o viral infection .
was determined for the compounds 1isted below.
';

, ,',, '' I
I
, , . I ' , .

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I!

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1++++++++++~+

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++++~ ~C ' I
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Claims (9)

THE EMBODIMENTS OF THE INVENTION IN WHICH AN EXCLUSIVE
PROPERTY OR PRIVILEGE IS CLAIMED ARE DEFINED AS FOLLOWS:

1. A process for preparing a compound of the formula V
and the pharmaceutically acceptable acid addition salts thereof, wherein R1 and R2 are each selected from the group consisting of normal alkyl of from 14 to 18 carbon atoms and octadec-9-enyl which comprises the steps of:
(a) reducing a compound of the formula XII; and (b) where required, converting the resulting compound of formula V to a pharmaceutically acceptable acid addition salt thereof.

2. A process of claim 1 wherein R1 and R2 are each normal alkyl of from 14 to 18 carbon atoms.

3. The process of claim 2 wherein R1 and R2 are each n-hexadecyl.

4. The process of claim 2 wherein R1 and R2 are each n-octadecyl.

5. A compound of the formula V or a pharmaceutically acceptable acid addition salt thereof, when prepared by the process of claim 1 or by an obvious chemical equivalent thereof.

6. A process for the preparation of 1-[2,3-di(n-hexadecyl-oxy)propyl]-4-aminomethyl-4-phenylpiperidine and its dihydro-chloride which comprises reducing 1-[2,3-di(n-hexadecyloxy)propyl]-4-cyano-4-phenylpiperidine with lithium aluminum hydride, and where required converting the free base to its dihydrochloride.

7. 1-[2,3-Di(n-hexadecyloxy)propyl]-4-aminomethyl-4-phenyl-piperidine and its dihydrochloride, when prepared by the process of claim 6 or by an obvious chemical equivalent thereof.

8. A process for the preparation of 1-[2,3-di(n-octadecyl-oxy)propyl]-4-aminomethyl-4-phenylpiperidine and its dihydro-chloride which comprises reducing 1-[2,3-di(n-octadecyloxy)propyl]-4-cyano-4-phenylpiperidine with lithium aluminum hydride, and where required converting the free base to its dihydrochloride.

9. 1-[2,3-Di(n-octadecyloxy)propyl]-4-aminomethyl-4-phenyl-piperidine and its dihydrochloride, when prepared by the process of claim 8 or by an obvious chemical equivalent thereof.