119)
`United States Patent
`5,595,980
`(11) Patent Number:
`
`[45] Date of Patent: Jan. 21, 1997
`Brodeetal.
`
`ACAA
`
`US00559
`
`(54)
`
`CONTRACEPTIVE COMPOSITIONS
`
`[75]
`
`Inventors: George L. Brode, Bridgewater, N.J.;
`Gustavo F. Doncel, Norfolk, Va.;
`Henry L. Gabelnick, N. Bethesda,
`Md.; Russell L. Kreeger, Flemington;
`.
`8/1986 European Pat. Off.
`0189935
`George A. Salensky, White House
`2078110==1/1982 United Kingdom .
`Station, both of N.J.
`OTHER PUBLICATIONS
`
`4,707,362 11/1987 NUWAYSET sss 424/433
`4,845,175
`7/1989 LO...
`wee 326/200
`4,929,722
`5/1990 Partain et ALL secserersnrvecvevntseee 536/20
`4,946,870
`8/1990 Partain et al. wees SLA/777
`
`
`
`FOREIGN PATENT DOCUMENTS
`
`(73]
`
`[21]
`
`[22]
`
`[63]
`
`(51)
`(52]
`
`[58]
`
`[56]
`
`Assignees: Medical College of Hampton Roads,
`Arlington, Va.; Biomaterials
`Corporation, Plainsboro, N.J.
`
`Appl. No.: 418,884
`
`Filed:
`
`Apr. 7, 1995
`
`Related U.S. Application Data
`
`Continuation of Ser. No. 129,253, Sep. 29, 1993, abandoned.
`
`Baits CU.
`ssscscesssssessosssesssssssssssessssss
`. AGIK 31/72
`U.S. Cl.....
`a "514/57;514/59; 514/55;
`
`514/814;514/843; 514/935; 514/944; 514/945;
`536/44; 536/55.1; 536/99; 424/DIG. 14
`
`Field of Search .
`. 514/814, 843,
`514/55,57,59,935,“944, 945; 536/99,
`44, 55.1; 424/DIG. 14
`
`References Cited
`
`U.S. PATENT DOCUMENTS
`
`4,228,277 10/1980 Landoll.....
`5/1983 Kazmiroski etal
`4,242,359 12/1980 Cooper etal. ....
`4,284,003
`
`424/329
`3/1982 Saurino .........00
`4,321,277
`
`424/180
`6/1983 Bagros ..
`4,387,094
`424/180
`7/1984 Maltz ....
`4,459,289
`4,474,769 10/1984 Smith ........
`1. 424/180
`
`4,551,148 11/1985 Riley et al.
`“
`...
`4,663,159
`5/1987 Brode ct al.
`scesssssensesnscnsesseun
`
`The Effects of Frequent Nonoxynol-9 Use On The Vaginal
`and Cervical Mucosa Somchai Niruthisard, MD etal., pp.
`176-179, 1991.
`Comparison of Vaginal Tolerance Test of Spermicidal Prepa-
`rations in Rabbits and Monkeys P. Eckstein etal., pp. 85-93,
`1969.
`HEC Cellosize Hydroxyethyl Cellulose-Union Carbide
`Corporation, pp. 1-32, 1991.
`
`Primary Examiner—Michael G. Wityshyn
`Assistant Examiner—Francisco C. Prats
`Attorney, Agent, or Firm—Banner & Witcoff Ltd.
`
`[57]
`
`ABSTRACT
`
`Improved contraceptive compositions are disclosed which
`comprise a spermicide or virucide, a polymeric delivery
`component and optionally a cosmetic ingredient. The
`improvementis directed to the use of certain hydrophobi-
`cally modified polysaccharides as the polymeric delivery
`component. Quite advantageously,
`the hydrophobically
`modified polysaccharides of the present invention can alter
`sperm motility. Moreover,
`the hydrophobically modified
`polysaccharides can provide reduced irritation potential
`when used in combination with spermicides such as, for
`example, nonoxynol-9, which may reduce the potential for
`infection of sexually transmitted diseases such as HIV and
`herpes.
`
`8 Claims, No Drawings
`
`Dr. Reddy's - EX1009
`Page 1
`
`Dr. Reddy's - EX1009
`Page 1
`
`

`

`5,595,980
`
`1
`CONTRACEPTIVE COMPOSITIONS
`
`This invention was made with government support under
`Cooperative Agreement DPE-3044-A-00-6063-00 between
`the United States Agency for International Development and
`the Medical College of Hampton Roads. The government
`has certain rights in this invention.
`This application is a continuation of application Ser. No.
`08/129,253, filed Sep. 29, 1993, now abandoned.
`
`FIELD OF THE INVENTION
`
`Thepeesent invention generally relates to contraceptive
`
`positions, and more specifically relates to improved
`
`give compositions comprising certain hydrophobi-
`
`caliy-medified polysaccharides as polymeric delivery com-
`ponents.
`
`BACKGROUND OF THE INVENTION
`
`Contraceptive compositions typically comprise an active
`ingredient, such as, for example, nonoxynol-9, a polymeric
`delivery component for delivering the active ingredient,
`such as, for example, hydroxyethyl cellulose or carboxym-
`ethyl cellulose, cosmetic ingredients, such as, for example,
`water, sorbitol and propylene glycol, and optionally other
`ingredients, such as, for example, stabilizers, fragrances,
`viscosity adjusters, and the like.
`One importantattribute of contraceptive compositions is
`that the active ingredients should be effective as a spermi-
`cide, In addition, the other ingredients present in the con-
`traceptive compositions should not interfere with the effec-
`tiveness
`of
`the
`active
`ingredient. Many
`existing
`contraceptive compositions possess these properties. How-
`ever, such existing contraceptive compositions typically do
`not have a high degree of substantivity to the mucosal lining
`of the vagina. Moreover, existing polymeric delivery com-
`ponents generally do not provide any functional effect with
`respect to altering sperm motility.
`Spermicides such as nonoxynol-9 and benzalkonium
`chloride have been used effectively as active ingredients in
`contraceptive compositions for many years. However,it has
`been found that such ingredients can be irritating to the
`mucosal lining of the vagina and cause an increased risk of
`vaginal irritation. Along with such increasedrisksof vaginal
`irritation, there may be increased risks of contracting sexu-
`ally transmitted diseases of bacterial, fungal or viral origin,
`such as, for example, HIV and herpes.
`Accordingly, improved contraceptive compositions are
`desired which are substantive and which can provide a low
`degree ofirritation to the mucosal lining of the vagina. In
`addition, improved contraceptive compositions are desired
`wherein polymeric delivery components are provided which
`can alter sperm motility.
`
`SUMMARYOF THE INVENTION
`
`In accordance with the present invention, improved con-
`traceptive compositions comprising a spermicide or viru-
`cide, a polymeric delivery component for the spermicide or
`virucide and cosmetic ingredients are provided wherein the
`polymeric delivery component comprises a hydrophobically
`modified polysaccharide. By virtue of the present invention
`it is now possible to provide contraceptive compositions
`wherein the polymeric delivery component can enhance
`effectiveness of the spermicide. As a result, the overall
`spermicidal effectiveness of the contraceptive compositions
`
`10
`
`20
`
`35
`
`50
`
`55
`
`2
`can be improved. In addition, the improved contraceptive
`compositions ofthe present invention are substantive to the
`mucosal lining of the vagina and can provide a reduced
`degree of vaginal
`irritation which may lower the risk of
`contracting sexually transmitted diseases.
`
`DETAILED DESCRIPTION OF THE
`INVENTION
`
`The contraceptive compositions of the present invention
`are suitable for use in mammals. As used herein, the term
`“mammals” means any class of higher vertebrates that
`nourish their young with milk secreted by mammary glands,
`e.g., humans, rabbits and monkeys.
`The spermicides useful in accordance with the present
`invention are known to those skilled in the art. Typical
`spermicides include, for example, benzalkonium chloride,
`octoxynol-9, ricinoleic acid, phenol mercuric acetates and
`nonoxynol-9, etc. Nonoxynol-9 and benzalkonium chloride
`are preferred spermicides for use in accordance with the
`present invention.
`The virucides suitable for use in the contraceptive com-
`positionsof the present invention are knownto thoseskilled
`in the art. Typical virucides include, for example, acyclovir,
`idoxyurnidine,
`ribavirin, nonoxynol-9, vidarabine
`and
`rimantadine.
`
`the contraceptive compositions of the present
`Thus,
`invention may typically comprise one or more spermicide or
`one or more virucide or both. Some ingredients such as, for
`example, nonoxynol-9, may function both as spermicide and
`virucide.
`
`Thetotal amount of spermicide and virucide, or mixtures
`thereof, will typically range from about 0.1 to 50 weight
`percent based on the weight of the contraceptive composi-
`tion. Preferably,
`the amount of spermicide or virucide
`employed will be that amount necessary to achieve the
`desired spermicidal or virucidal
`results. Appropriate
`amounts can be determined by those skilled in the art.
`Preferably,
`the total concentration of the spermicide or
`virucide, or mixtures thereof, will comprise from about1 to
`25 weight percent, and more preferably from about 1 to 5
`weight percent, based on the weight of the contraceptive
`composition.
`The polymeric delivery componentssuitable for use in the
`contraceptive compositions of the present invention com-
`prise one or more hydrophobically modified polysaccharides
`selected from the group consisting of cellulosics and chito-
`sans. Such polysaccharidestarting materials from which the
`hydrophobically modified polysaccharides of the present
`invention can be made are known to those skilled in the art.
`Typical cellulosics include, for example, hydroxyethyl cel-
`lulose, hydroxypropyl] cellulose, methy] cellulose, hydrox-
`ypropylmethyl cellulose, hydroxyethyl methyl cellulose,
`and the like. Preferred cellulosics include hydroxyethyl
`cellulose and hydroxypropyl cellulose. Typical chitosans —
`include, for example, the following chitosan salts; chitosan
`lactate, chitosan salicylate, chitosan pyrrolidone carboxy-
`late, chitosan itaconate, chitosan niacinate, chitosan formate,
`chitosan acetate, chitosan gallate, chitosan glutamate, chi-
`tosan maleate, chitosan aspartate, chitosan glycolate and
`quaternary amine substituted chitosan and salts thereof,etc.
`Chitosan lactate and chitosan pyrrolidone carboxylate and
`are preferred chitosans. The polymeric delivery component
`may comprise mixtures of polysaccharides between classes
`of the group,e.g,, cellulosics and chitosans,or within a class,
`e.g., two cellulosics.
`
`Page 2
`
`Page 2
`
`

`

`5,995,980
`
`3
`The hydrophobically modified polysaccharides of the
`present invention comprise a hydrophobic substituent con-
`taining a hydrocarbon group having from about 8 to 18
`carbon atoms, preferably from about 10 to 18 carbon atoms
`and more preferably from about 12 to 15 carbon atoms. The
`hydrocarbon group of the hydrophobic substituent may
`comprise an alkyl or arylalkyl configuration. As used herein
`the term “arylalkyl group” means a group containing both
`aromatic and aliphatic structures. Procedures for hydropho-
`bically modifying the above mentioned polysaccharides are
`knownto those skilled in the art. See, for example, U.S. Pat.
`Nos. 4,228,277 issued Oct. 14, 1980 and 4,663,159 issued
`May5, 1987.
`The degree of substitution of the hydrophobic substituent
`on the polysaccharide is typically from about 0.05 to 0.5,
`preferably from about 0.08 to 0.25, more preferably 0.08 to
`0.16 and most preferably from greater than about 0.11, e.g.,
`0.12, to less than 0.16, e.g., 0.15, moles of the hydrophobic
`substituent per mole of polysaccharide. The hydrophobic
`substituent may be anionic, cationic, nonionic or amphot-
`eric. More than one particular hydrophobic substituent can
`be substituted onto the polysaccharide provided that the total
`substitution level] is within the ranges set forth above.
`A preferred hydrophobic substituent is a cationic, quater-
`nary, nitrogen-containing radical having the formula:
`
`R,
`
`RoHOR (Ai?)
`R3
`
`wherein:
`
`each R, and R, are CH, or C,H;
`R, is CH,CHOHCH,or CH,CH,;
`R, is an alkyl or arylalkyl group having about 8 to 18
`carbon atoms; and
`A, is a halide ion.
`Preferably, R, and more preferably, both R, and R, are
`CH,. Preferably, R, is CH,CHOHCH,. Preferably, R, is
`C,H(2n+1), where n is from 8 to 18. An especially preferred
`hydrophobicgroup,i.e., Ry, has the formula C,,H,,. Chlo-
`rine is a preferred halide ion.
`Other preferred hydrophobic substituents include those
`prepared from hydrophobe containing reagents such as
`glycidyl ethers, e.g., nonylphenylglycidyl ether or dode-
`cylphenylglycidyl ether, alphaolefin epoxides, e.g., 1,2
`epoxy hexadecane andtheir respective chlorohydrins, alkyl
`halides, e.g., dodecylbromide, and mixtures thereof.
`The ionic character of the hydrophobically modified
`polysaccharides of the present invention is not critical and
`can be anionic, cationic, nonionic or amphoteric. However,
`cationic polysaccharides are preferred for use in accordance
`with the present invention. Thus, in a preferred aspect ofthe
`invention, the polysaccharides are also substituted with an
`ionic substituent in addition to the hydrophobic substituent.
`The amountoficnic substituent typically ranges from about
`0.05 to 0.9, preferably from 0.10 to 0.25, moles of the ionic
`substituent per mole of the polysaccharide for cellulosics
`and preferably from about 0.5 to 0.9 moles of the ionic
`substituent per mole of the polysaccharide for chitosan
`derivatives. More than one particular ionic substituent can
`be substituted onto the polysaccharide providedthat the total
`substitution level is within the ranges set forth above.
`A preferred cationic substituent for cellulosics is a cat-
`ionic quaternary nitrogen containing radical having the
`formula:
`
`4
`
`*
`Ro“WOR [A2®]
`Ry
`
`10
`
`20
`
`25
`
`30
`
`35
`
`45
`
`wherein
`
`each R,, R, and R, are CH, or C,H,;
`R, is CH, CHOHCH, or CH,CH,; and
`Azis a halide ion.
`Preferably, at
`least one of R;, Rg and R, are CH.
`Preferably R, is CH,CHOHCH,,.Preferably, A, is a chloride
`anion,
`
`A preferred cationic substituent for chitosans is an ammo-
`nium group containing radical having the formula:
`
`NH,@ A,®
`
`wherein A, is an organic acid counter ion. Preferably, A, is
`lactate pyrrolidone carboxylate, acetate or combinations
`thereof.
`In addition to the above described hydrophobically modi-
`fied polysaccharides, the contraceptive compositions may
`contain other polysaccharides, or deriyatives thereof, such
`as,
`for example; hydroxyethyl celiufose, carboxymethyl
`cellulose, dextran sulfate and hyaluronic acid. Such other
`polysaccharides may or may not be hycrophobically modi-
`fied. Such other polysaccharides, when present in the com-
`position, may comprise from about 0.1 to 25%, based on the
`weight of the contraceptive compositions. One preferred
`contraceptive composition in accordance with the present
`invention comprises a cationic hydrophobically modified
`hydroxyethyl cellulose in combination with chitosan lactate
`as the polymeric delivery component.
`Preferably, the hydrophobically modified polysaccharides
`of the present invention are water soluble. As used herein,
`the term “water soluble” means that at least
`1 gram and
`preferably at least 2 grams of the hydrophobically modified
`polysaccharideare soluble in 100 gramsofdistilled water at
`25° C. and 1 atmosphere. The degree of water solubility can
`be controlled by varying the amount of ether substitution,
`hydrophobe substitution and cation substitution on the
`polysaccharide,
`the details of which are known to those
`skilled in the art.
`The molecular weight of the polysaccharides suitable for
`use in accordance with the presentinvention typically ranges
`from about 10,000 to 500,000 grams per gram mole and
`preferably ranges from about 20,000 to 200,000 grams per
`gram mole. As used herein, the term “molecular weight”
`means weight average molecular weight. Methodsfor deter-
`mining weight average molecular weight of polysaccharides
`are knownto those skilled in the art. One preferred method
`for determining molecular weight is low angle laser light
`scattering. The viscosity of the polysaccharides typically
`ranges from about 5 to 5000 centipoise, preferably from
`about 10 to 500 centipoise. Unless otherwise indicated, as
`used herein the term “viscosity” refers to the viscosity of a
`2.0 weight percent aqueous solution of the polymer mea-
`sured at 25° C. with a Brookfield viscometer. Such viscosity
`measuring techniques are knownto those skilled in the art.
`Typically, the amount of the polymeric delivery compo-
`nent will range from about 0.1 to 99.9 weight percent,
`preferably, from about 0.5 to 50 weight percent and more
`preferably from about 1 to 10 weight percent, based on the
`weight on the contraceptive composition.
`The balance of the contraceptive compositions of the
`presentinvention,i.e., typically from about 0.1 to 99.8% and
`
`Page 3
`
`Page 3
`
`

`

`5,595,980
`
`5
`often about 50 to 99.8 weight percent, may optionally
`comprise one or more cosmetic ingredients. Such cosmetic
`ingredients are known to those skilled in the art and are often
`referred to in the art as diluents, solvents and adjuvants.
`Typically cosmetic ingredients include, for example; water,
`ethyl alcohol, isopropyl alcohol, glycerin, glycerol propy-
`lene glycol, sorbitol and other high molecular weight alco-
`hols. In addition, contraceptive compositions of the present
`invention may contain minor amounts, e.g. from about 0.1 to
`5% weight based on the weight of the contraceptive com-
`positions, of other additives, such as, for example; stabiliz-
`ers, surfactants, menthol, eucalyptusoil, other essential oils,
`fragrances, and the like. Polyoxyethylene 20osorbitan
`monolaurate is a preferred stablizer for use in the compo-
`sitions of the present invention. In fact, in accordance with
`the present invention, it is believed that certain stabilizers,
`such as, for example, polyoxyethylene 20-sorbitan mono-
`laurate, may contribute to the sperm blocking properties of
`the hydrophobically modified polysaccharides of the present
`invention. Details concerning the selection and amounts of
`cosmetic ingredients, other additives, and blending proce-
`dures are knownto those skilled in the art.
`The contraceptive compositions of the present invention
`may be delivered to the vagina of a mammal by any means
`knownto those skilled in the art. Typical forms for delivery
`of the compositions include, for example; creams, lotions,
`gels, foams, sponges, suppositories andfilms. In addition the
`compositions of the present invention may be used as
`personal care lubricants, such as, for example, as condom
`lubricants, and the like. Such lubricants may comprise
`commonly knowningredients such as, for example: humec-
`tants; e.g., glycerine, sorbitol, mannitol, glycols and glycol
`ethers; buffers, e.g., glucono-d-lactone; germicides or bac-
`tericides, e.g., chlorhexidine gluconate; preservatives, e.g.,
`methylparaben; viscosifiers; e.g., hydroxyethyl cellulose,
`etc.; other adjuvents; e.g., colors and fragrances;in addition
`to the compositions of the present invention. Those skilled
`in the art will recognize that the physical properties, e.g.,
`viscosity, of such delivery forms may vary widely. For
`example, the viscosity of a gel form of the composition of
`the present invention, e.g. 150,000 centipoise, may be sub-
`stantially higher than the viscosity of lotion form of the
`composition of the present invention, e.g., 100 centipoise.
`Further details concerning the materials, ingredients, pro-
`portionsand proceduresof such delivery forms are known to
`those skilled in the art.
`The contraceptive compositions of the present invention
`are preferably administered to the vagina of the mammal in
`a dosage whichis effective to immobilize sperm present in
`the vagina and/or to inhibit their penetration in cervical
`mucus. Typical dosages range between about 0.01 to 0.2
`gramsof the composition per kilogram of body weightof the
`mammal.
`Quite surprisingly, it has been found that the hydropho-
`bically modified polysaccharides of the present invention
`can provide a high degree of substantivity to the mucous
`membrane ofthe vagina, in addition to being non-irritating
`to the mucous membrane even in the presence of normally
`irritating active ingredients such as Nonoxonyl-9, Moreover,
`the hydrophobically modified polysaccharides of the present
`invention can provide a high degreeofsaline compatibility.
`Saline compatability is an importantattribute of contracep-
`tive compositions. As usedherein, the term “saline compat-
`ability” means that the contraceptive composition remains
`dissolved, i.e., does not separate at 25° C. and ] atmosphere,
`in a saline solution, i.e., 9 grams of NaCl per liter of water,
`at concentrations of up to at least 2 weight percent, prefer-
`
`6
`ably 5 weight percent, for at least one hour, preferably at
`least 24 hours. Preferably, there are appropriate levels of the
`hydrophobic substituentandthe ionic substituent to enhance
`the saline compatibility of the composition. The molar ratio
`of the ionic substituent to the hydrophobic substituent is
`preferably at least 1.5:1, more preferably 2.0:1 and most
`preferably at least 2.5:1. When the hydrophohic substituen)..
`is not ionic, the molar ratio of the ionic sut#gigient to the
`hydrophobic substituent is equal to the molar ratio of the
`ionic substituent to the hydrophobic substituent. When the
`hydrophobic substituentis ionic, the molar ratio of the ionic
`substituent to the hydrophobic substituent is equal to the
`sum of the moles of ionic substituents and hydrophobic
`substituents per mole of hydrophobic substituent. For
`example,if the substitution level of a cationic, hydrophobic
`substituent is 0.12 gram moles per gram mole of polysac-
`charide, and the substitution level of the cationic substituent
`is 0.2 gram moles per mole of polysaccharide,then the molar
`ratio of the ionic substituent to the hydrophobic substituent
`would be 2.67,i.e., (0.12+0.20)+0.12=2.67.
`Thus, the compounds of the present invention are par-
`ticularly suitable for use as excipients for contraceptive
`compositions because of their desirable combination of
`saline compatibility, low irritation potential, substantivity
`and ability to impair sperm motility.
`
`20
`
`EXAMPLES
`
`30
`
`The following examples are provided for illustrative
`purposesandare not intendedto limit the scope of the claims
`which follow.
`
`DEFINITIONS
`
`The following ingredients were used in the Examples.
`CMC—carboxymethyl cellulose having a viscosity of
`400-800 centipoise, available from Aqualon Company,
`Wilmington, Del.
`CL—chitosan lactate having a 1% solution viscosity of 15
`to 250 centipoise, available from Dainichiseika Colors and
`Chemicals Co. Ltd., Tokyo, Japan.
`CONCEPTROL—a commercially available contracep-
`tive composition containing CMC and POV sold by
`Advanced Care Products, Ortho, Johnson and Johnson, New
`Brunswick, N.J.
`trimethyl ammonium chloride
`CS1—2,3 epoxypropyl
`available from DeGussa Corporation, sold as Quab 151.
`DS—dextran sulfate having a molecular weight of
`40,000-50,000 g/gmole, available from United States Bio-
`medical Corp., Cleveland, Ohio.
`HECi1—hydroxyethyl cellulose having a viscosity of
`4400-6000 centipoise (1% solution) available from Union
`Carbide Corp., Danbury, Conn., sold as Cellosize® QP-100
`M.
`
`HPC1—hydroxypropyl cellulose having a viscosity of
`1500-3000 centipoise (1% solution) available from Aqualon
`Company, Wilmington, Del.
`dimethyldodecyl
`HS1—3-chloro-2-hydroxypropy]
`ammonium chloride available from DeGussa Corporation,
`Ridgefield Park, N.J., sold as Quab 342.
`dimethyloctadecyl
`HS2—3-chloro-2-hydroxypropyl
`ammonium chloride available from DeGussa Corporation,
`Ridgefield Park, N.J., sold as Quab 426.
`HS3—nonylphenylglycidyl ether available from Rhone
`Poulenc sold as Heloxy 64.
`
`50
`
`65
`
`Page 4
`
`Page 4
`
`

`

`5,595,980
`
`7
`JR—a cationic hydroxyethyl cellulose having a viscosity
`of 300-500 centipoise available from Union Carbide Corp.,
`Danbury, Conn.
`N-9—Nonoxynol-9 USP available from Rhone Poulenc,
`Cranberry, N.J., sold as Igepal CO-630 Special.
`P-20—polyoxyethylene 20-sorbitan monolaurate, avail-
`able from ICI Americas, Inc., Wilmington, Del., sold as
`Tween 20.
`
`P-80—polyoxyethylene 80-sorbitan mono-oleate, avail-
`able from ICI Americas, Inc., Wilmington, Del., sold as
`Tween 80.
`
`PG—propylene glycol USP,available from Fisher Scien-
`tific, Fairlawn, N.J.
`POL. 1—acationic, hydrophobically modified hydroxy-
`ethyl cellulose having a viscosity of 100 to 500 centipoise
`(2% solution) and containing a hydrophobic substituent
`containing a hydrocarbon portion having 12 carbon atoms
`and a cationic substituent, available from Union Carbide,
`Danbury, Conn. sold as Quatrisoft®.
`POL. 2—a cationic, hydrophobically modified hydroxy-
`ethyl cellulose having a viscosity of 50 to 500 (2% solution)
`centipoise and containing a hydrophobic substituent con-
`taining a hydrocarbon portion having 12 carbon atoms and
`a cationic substituent.
`
`POL. 3—a cationic, hydrophobically modified hydroxy-
`ethyl cellulose having a viscosity of 50 to 500 (2% solution)
`centipoise and containing a hydrophobic substituent con-
`taining a hydrocarbon portion having 18 carbon atoms and
`a cationic substituent.
`
`POL. 4—a non-ionic hydrophobically modified hydroxy-
`ethyl cellulose having a molecular weight of 300,000
`g/gmole having a hydrophobic substituent containing a
`hydrocarbon portion having 16 carbon atoms available from
`the Aqualon Company, Wilmington, Del., sold as Natrosol®
`Plus.
`
`POL. 5—a hydrophobically modified dextran sulfate hav-
`ing a molecular weight of 50,000 g/gmole and containing
`2.8 wt. % of a hydrophobic substituent containing a hydro-
`carbon portion having 15 carbon atoms.
`POL. 6—a hydrophobically modified carboxymethy! cel-
`lulose having a viscosity of 50 to 500 (2% solution) centi-
`poise and containing 1.2 wt. % of a hydrophobic substituent
`containing a hydrocarbon portion having 15 carbon atoms.
`POV—polyvinyl pyrrolidone Povidone USP having a
`molecular weight of 45,000 g/gmole, available from ISP
`Chemicals Wayne, N.J.
`SOR—sorbitol, available from Fisher Scientific, Fair-
`lawn, N.J,
`The following tests were used in the Examples.
`Modified One End Test (MOET)—This test was used to
`determine the effect of various compounds on sperm pen-
`etration in cervical mucus. Capillary tubes containing
`bovine cervical mucus obtained from Serono-Baker Diag-
`nostics Inc., Allentown, Pa. sold as Penetrax, were used to
`conductthe test. Each of the test compositions containing
`the polymer to be tested was diluted in a saline solution,i.e.,
`at 9 grams of NaC!per liter of water, to a polymer concen-
`tration of between 0.007 w/v % and 0.45 wiv % (wiv %
`equals grams per 100 milliliters). The test was conducted at
`a concentration of either 0.003 w/v% polymer, 0.007 w/v %
`polymer or 1 g of test composition per 11 ml of saline. The
`tubes were thawedbriefly and then broken open. The open
`end was placed in a container containing the sample in
`saline. The sample was allowed to migrate for 30 minutes
`through the tube. A semen sample was then diluted with a
`
`15
`
`20
`
`25
`
`30
`
`35
`
`45
`
`50
`
`55
`
`60
`
`65
`
`8
`buffer solution to 60 million motile sperm per milliliter and
`mixed with the polymer sample. The tube containing the
`polymer sample was then re-inserted into the container
`containing the mixed solution and stored in an incubator at
`37° C. in an atmosphere of 5 percent carbon dioxide in air
`for 60 minutes. The container and tube were then removed
`
`from the incubator and the tube wasvisually analyzed under
`a microscope for the migration of motile vanguard sperm
`through the tube. The results are expressed as percentage of
`migration as compared to control samples. In the control
`samples, the tubes were incubated with saline containing no
`polymer.
`Double End Test (DET)—This test was also used to
`biologically evaluate the diffusion of the compounds in
`cervical mucus. The DET is similar to the MOET with the
`exception that 20 millimeter capillary tubes were exposed to
`the polymer samples by one end for 60 minutes and subse-
`quently by the other end to the semen solution for 60
`minutes so that sperm could migrate in the opposite direc-
`tion of the polymer sample. Penetration length of vanguard
`motile sperm is recorded and the results are expressed as
`percentage of migration as compared to control samples, i.e.,
`saline containing no polymer. The shorter the sperm pen-
`etration, the greater the compoundbiodiffusion. In addition,
`the samples used for the DET were further modified to
`contain 4 weight percent of N-9. The DET valuesreflect how
`far a test compound can physically diffuse in cervical mucus
`while still displaying sperm penetration inhibitory activity.
`Simultaneous One End Test (SOET)—This test was used
`to detect
`the quick blocking effects of the compounds
`particularly exerted through sperm motility alterations. The
`SOET is similar to the MOET except that the solution
`containing the polymer is mixed with the semen sample and
`then oneendofthe capillary tube containing bovine cervical
`mucusis inserted into the mixture of the polymer and semen
`sample andstoredin an incubatorat 37° C. in an atmosphere
`of 5% carbon dioxide in air for 60 minutes. Penetration
`length of vanguard motile sperm is recorded and the results
`are expressed as percentage of migration as compared to
`control samples, i.c., saline containing no polymer. In the
`SOET,if not impeded by the test compound, the sperm have
`the ability to migrate into the tube immediately after contact.
`Sander-Cramer test—This test was used to evaluate the
`spermicidal effectiveness of contraceptive compositions.
`The Sander-Cramertest was developedin the laboratories of
`Ortho Pharmaceutical Corporation. A slight modification of
`the original protocol was used as described below. Serial
`dilutions of each test composition in volumes of 250 micro-
`liters were added to 50 microliters of semen adjusted to 60
`million motile sperm per milliter at room temperature. The
`end point was thegreatest. dilution at which all of the sperm
`were immobilized within 20 seconds. Results are expressed
`as minimum effective concentrations in milligrams per mil-
`liliter.
`
`Example 1
`
`Preparation of Cellulose Ether Derivative
`
`A reaction vessel equipped with a stirrer, condenser,
`addition funnels, and nitrogen supply, was charged with 39
`grams of HEC] and 272 grams of anhydrous acetone. The
`reactor was purged with nitrogen and 23 grams of an
`aqueous sodium hydroxide solution containing 20 wt %
`sodium hydroxide was added. After stirring for 30 minutes,
`64 g of an aqueous solution containing 40 wt % HS1 was
`added. The reactor mixture was heated to 55° C. and held
`
`Page 5
`
`Page 5
`
`

`

`5,595,980
`
`10
`9
`Thereafter, 2.62 grams were dissolved in 97.38 grams of
`there for 2 hours. Then 8.7 grams of an aqueous solution
`containing 70 wt % CS1 was added. The mixture was held
`water with good agitation and heated to 75° C. for complete
`at 55° C. for another hour. The reaction was cooled and
`solution. The pH of the solution was adjusted to 4.7 with
`neutralized with 3 grams glacial acetic acid. The reaction
`lactic add. The solution viscosity as determined by a Brook-
`.
`slurry wasfiltered and washed 7 times with 400 grams of an 5
`‘ld Cone & Plate Viscometer Model DV-1 CP-41 at 20 rpm
`aqueous solution containing 90 wt % acetone, once with 400
`grams of an aqueoussolution containing 94 wt % acetone,|Was 609 centipoise. The 2.5 wt. % solution was thenused to
`and once with 400 grams of a solution containing 0.5
`make test compositions with varying polymer solids content
`milliliter of a 40 wt % glyoxal solution, 0.5 milliliter of
`by dilution with water.
`acetic acid and the balance acetone. After drying, 58 grams
`2) A test compositi
`taining
`1.25 wt. % polymer and
`of product containing 1.5% volatiles was obtained. The
`i Oe by >00aoe eretbemca 50 ee of the
`nitrogen content of the polymer was 1.60 wt. %, and the
`WE
`®Y
`BN
`8)
`“<0
`prepared
`polymer had a 2% solution viscosity of 190 centipoise.
`above 2.5 wt. % solution (1) with 1.25 grams of P-20 and
`48.75 grams of water. The MOET was then performed using
`15 a sample containing1 g of the test composition per 11 ml
`Example 2
`saline. The MOETvalue was 0% indicating an exceptional
`cn ae
`reduction in sperm penetration in cervical mucus. This
`Nonoxynol-9 Compatibility
`finding is particularly important since the same sample
`In order to compare the compatibility of nonoxynol-9
`with non-hydrophobically modified polymeric delivery . failed to completely immobilize sperm underthe conditions
`compo and thepolymeric oooanole.taeconofthe
`employed in the Sander-Cramertest. The MOETvalue was
`present
`invention,
`ie
`TOWOWwINg Cx
`- Was condu
`“
`:
`5a
`HEC1, HPC1 and POL.4 were used in this Example.
`12% ata polymer concentration of 0.007 wiv% also ind
`:
`os
`cating an exceptional reduction in sperm penetration in
`One hundred gram aqueous solutions containing 1.5
`ical
`weight percent of the polymeric delivery component being ,;
`cervical mucus.
`tested were prepared. To each solution, 4,16 grams of N-9
`(3) An additional quantity of the above test composition
`were added and mixed for 20 minutes. Each solution was
`was used to prepare a 4 wt. % N-9 composition containing
`then divided into two portions. The first portion wasstored
`1.25 wt. % polymer and 1.25 wt. % P-20, Approximately 96
`at room temperature, i.¢., about 25° C., and the second
`gramsofthe test composition described in (2) above was
`solution was stored at about 35° C. After 24 hours, the 4,
`—.
`d
`with 4.0
`f N-9 for 30 minutes. The DET was
`
`solution containing HEC1 demonstrated a phase separation~XC WHS Erams oO! N= ures. 7 was
`
`
`
`which indicated that the HEC1, which was not hydropho-
`__then performedand the result showeda sperm penetration of
`bically modified, was incompatible with N-9. Similarly,
`60%. This reveals a compound cervical mucusbiodiffusion
`after 24 hours t