Case 1:17-cv-00868-JFB-SRF Document 13-7 Filed 08/18/17 Page 1 of 11 PageID #: 268
`Case 1:17-cv—00868—JFB-SRF Document 13-7 Filed 08/18/17 Page 1 of 11 PageID #: 268
`
`
`
`
`
`EXHIBIT 7
`
`EXHIBIT 7
`
`

`

`Case 1:17-cv-00868-JFB-SRF Document 13-7 Filed 08/18/17 Page 2 of 11 PageID #: 269
`
`US009018177B2
`
`(12) United States Patent
`Lauten et al.
`
`(10) Patent No.:
`(45) Date of Patent:
`
`US 9,018,177 B2
`Apr. 28, 2015
`
`(54) COSMETIC COMPOSITIONS FOR
`INCREASING BIOAVAILABILITY OF THE
`ACTIVE COMPOUNDS BAICALIN AND/OR
`VITAMIN C
`
`(71) Applicant: L'Oreal S.A., Paris (FR)
`
`(72) Inventors: Elizabeth Hunter Lauten, New York,
`NY (US); Jean-Thierry Simonnet,
`Mamaroneck, NY (US); Zhi Pan, Fort
`Lee, NJ (US); Guive Balooch, New
`York, NY (US)
`(73) Assignee: L'Oreal S.A., Paris (FR)
`(*) Notice:
`Subject to any disclaimer, the term of this
`patent is extended or adjusted under 35
`U.S.C. 154(b) by 98 days.
`(21) Appl. No.: 13/650,933
`
`(22) Filed:
`
`Oct. 12, 2012
`
`(65)
`
`Prior Publication Data
`|US 2014/01 07047 A1
`Apr. 17, 2014
`
`(2006.01)
`(2006.01)
`(2006.01)
`(2006.01)
`(2006.01)
`(2006.01)
`(2006.01)
`(2006.01)
`
`(51) Int. Cl.
`A6 IK 9/{}{}
`A6 IK 47/12
`A6 IK 47/22
`A6 IK 3.1/7048
`A6 IK 8/368
`A6 IK 8/49
`A6 IK 8/67
`A610 19/00
`(52) U.S. CI.
`CPC ............... A61K 9/0014 (2013.01); A61K 47/12
`(2013.01); A61K 47/22 (2013.01); A61K
`31/7048 (2013.01); A61K 8/368 (2013.01);
`A61K 8/4913 (2013.01); A61K 8/4953
`(2013.01); A61K 8/498 (2013.01); A61K 8/675
`(2013.01); A61K 8/676 (2013.01); A610 19/00
`(2013.01); A61K 2800/10 (2013.01)
`(58) Field of Classification Search
`CPC ....... A61K 47/00; A61K 8/4973; A61K 8/60;
`A61K 31/7032; A61C) 19/00; A61O 90/00
`See application file for complete search history.
`
`(56)
`
`References Cited
`
`|U.S. PATENT DOCUMENTS
`
`3/1970 Matson
`3,501,409 A
`3,839,210 A 10/1974 Beiswanger et al.
`4,680,143 A
`7/1987 Edge et al.
`5,532,012 A
`7/1996 Balentine et al.
`5,686,082 A 11/1997 N’Guyen
`5,900,416 A * 5/1999 Markson ....................... 424/728
`6,121,209 A
`9/2000 Watts et al.
`
`6,331,520 B1 12/2001 Richardson
`6,355,657 B1
`3/2002 Osborne
`6,423,327 B1
`7/2002 Dobson, Jr. et al.
`6,479,442 B1 11/2002 Berube et al.
`6,645,513 B2 11/2003 Dobson, Jr. et al.
`6,646,035 B2 11/2003 Koch et al.
`6,733,797 B1
`5/2004 Summers
`6,949,496 B1
`9/2005 Boutique et al.
`7,452,549 B2 11/2008 Hasler-Nguyen et al.
`2002/0086042 A1
`7/2002 Delrieu et al.
`2002/0110604 A1
`8/2002 Babish et al.
`2003/0031715 A1
`2/2003 Park et al.
`2003/0206972 A1
`11/2003 Babish et al.
`7/2004 Galey
`2004/0146474 A1
`2005/0158271 A1
`7/2005 Lee et al.
`2005/0266.121 A1
`12/2005 Lines et al.
`2006/01 10439 A1
`5/2006 Tobia et al.
`9/2007 Lipshutz
`2007/0208088 A1
`10/2007 Leung et al.
`2007/0232561 A1
`4/2008 Declercq et al.
`2008/0095866 A1
`2008/0176956 A1
`7/2008 HSu
`2008/0219927 A1
`9/2008 Thakur et al.
`2009/0110674 A1
`4/2009 Loizou
`2009/023.3876 A1
`9/2009 Auriol et al.
`2010/0047297 A1
`2/2010 Petersen
`2011/0033525 A1
`2/2011 Liu
`3/2011 Ng et al.
`2011/0067294 A1
`2011/0136245 A1
`6/2011 Parker
`2011/0152214 A1
`6/2011 Boison et al.
`2012/007 1550 A1
`3/2012 Zelkha et al.
`
`FOREIGN PATENT DOCUMENTS
`
`6150918 A * 3/1986 ............. A61K 3 1/35
`JP
`WO WO-20 130 16257 A1
`1/2013
`
`OTHER PUBLICATIONS
`
`Mou, H., Wang, X., Lv, T., Xie, L., & Xie, H. (2011). On-line
`dissolution determination of Baicalin in solid dispersion based on
`near infrared spectroscopy and circulation dissolution system.
`Chemometrics and Intelligent Laboratory Systems, 105(1), 38-42.*
`Makoto et al., JP 6150918 A, Mar. 1986, machine translation of
`abstract, http://worldwide.espacemet.com, and Derwent record from
`EAST. Retreived on Mar. 19, 2014.*
`Suzuki, H. et al., “Mechanistic Studies on Hydrotropic Solubilization
`of Nifedipine in Nicotinamide Solution.” Chem. Pharm. Bull. 46(1),
`125-130 (1998).
`
`(Continued)
`
`Primary Examiner – Shaojia Anna Jiang
`Assistant Examiner – Dale R Miller
`(74) Attorney, Agent, or Firm – Novak Druce Connolly
`Bove + Quigg LLP
`
`ABSTRACT
`(57)
`Aqueous compositions comprisinga) at least one active com
`pound and b) at least one hydrotrope in an amount effective to
`increase transdermal penetration of the active compound, and
`methods for increasing bioavailability of active compounds
`are provided.
`
`14 Claims, 3 Drawing Sheets
`
`

`

`Case 1:17-cv-00868-JFB-SRF Document 13-7 Filed 08/18/17 Page 3 of 11 PageID #: 270
`
`US 9,018,177 B2
`Page 2
`
`(56)
`
`References Cited
`
`OTHER PUBLICATIONS
`
`Evstigneev, M.P. et al., “Effect of a mixture of caffeine and
`nicotinamide on the solubility of vitamin (B2) in aqueous solution.”
`European Journal of Pharmaceutical Sciences 28, 59-66 (2006).
`Da Silva, R.C. et al., “Investigations on the mechanism of aqueous
`solubility increase caused by some hydrotropes.” Thermochimica
`Acta 328, 161-167 (1999).
`Huh, K.M. et al., “A new hydrotropic block copolymer micelle sys
`tem for aqueous solubililzation of paclitaxel.” Journal of Controlled
`Release 126, 122-129 (2008).
`
`Takahashi, K. et al., “Application of hydrotropy to transdermal for
`mulations: hydrotropic solubilization of polyol fatty acid monoesters
`in water and enchancement effect on skin permeation of 5-FU.”
`Journal of Pharmacy and Pharmacology 63, 1008-1014 (2011).
`Nicoli, S. et al., “Association of nicotinamide with parabens: Effect
`on solubility, partition and transdermal permeation.” European Jour
`nal of Pharmaceutics and Biopharmaceutics 69, 613-621 (2008).
`Nidhi, K. et al., “Hydrotropy: A Promising Tool for Solubility
`Enhancement: A Review.” International Journal of Drug Develop
`ment & Research 3(2), 26-33 (2011).
`* cited by examiner
`
`

`

`Case 1:17-cv-00868-JFB-SRF Document 13-7 Filed 08/18/17 Page 4 of 11 PageID #: 271
`Case 1:17-cv-00868-JFB-SRF Document 13-7 Filed 08/18/17 Page 4 of 11 PageID #: 271
`
`US. Patent
`
`Apr. 28, 2015
`
`Sheet 1 013
`
`US 9,018,177 B2
`
`FIG 1.
`
`2
`
`bi?)
`
`lipidbliayer
`
`Hydrotropic
`Agent
`+
`Active
`
`‘
`‘ 1,!
`
`a
`
`.;'
`
`°
`
`glucose
`
`
`
`5% sodium saiicylate
`
`3% sodium saiicylate
`1% caffeine
`
`5% niacinamide +5°o caffeine
`
`5% niacinamide +5% caffeine + 5% urea
`
`3% sodium PCA +3% urea
`
`5% sodium PCA +3% urea
`
`5% sodium PCA +5% urea
`
`3% niacinamide +5% caffeine
`
`5% sodium PCA
`
`

`

`Case 1:17-cv-00868-JFB-SRF Document 13-7 Filed 08/18/17 Page 5 of 11 PageID #: 272
`
`U.S. Patent
`
`Apr. 28, 2015
`
`Sheet 2 of 3
`
`US 9,018,177 B2
`
`0.9
`
`0.8
`
`07
`
`(), 6
`
`0 5
`
`0. 4
`
`0.3
`
`0.2
`
`0,1
`
`0
`300
`
`Glucose Release after Presence of Hydrotrope
`-Reagent Blank
`
`* Glucose release after 5%
`niaCinámide +5% Caffeine
`* Glucose release after 5%
`U?ed
`- Control filtrate- no
`hydrotropepresent
`
`*&s.
`…
`
`%,
`º
`
`º \
`º *.
`*s, \
``s N. *...*,
`º
`
`
`
`310
`
`320
`
`330
`
`340
`
`350
`wavelength (nm)
`
`360
`
`370
`
`380
`
`390
`
`400
`
`FIG. 2B
`
`

`

`Case 1:17-cv-00868-JFB-SRF Document 13-7 Filed 08/18/17 Page 6 of 11 PageID #: 273
`
`U.S. Patent
`
`Apr. 28, 2015
`
`Sheet 3 of 3
`
`US 9,018,177 B2
`
`vine
`
`
`
`2 hr.
`
`6 hr
`
`16 hr
`
`

`

`Case 1:17-cv-00868-JFB-SRF Document 13-7 Filed 08/18/17 Page 7 of 11 PageID #: 274
`
`US 9,018,177 B2
`
`1
`COSMETIC COMPOSITIONS FOR
`INCREASING BIOAVAILABILITY OF THE
`ACTIVE COMPOUNDS BAICALIN AND/OR
`VITAMIN C
`
`FIELD OF THE INVENTION
`
`The present invention relates to aqueous compositions
`comprising at least one active molecule and at least one
`hydrotrope for increasing the bioavailability of the active
`molecule for topical applications.
`
`10
`
`BACKGROUND OF THE INVENTION
`
`The cosmetic market has begun to include many active
`ingredients in formulations to help curb the effect of aging
`and skin damage. Unfortunately, the efficacy of some of these
`molecules is reduced due to the natural barrier properties of
`the skin membrane. In particular the outer most stratum cor
`neum layer shows poor skin permeability of compounds that
`are hydrophilic, very lipophilic, of high molecular weight or
`charged.
`For example, transdermal penetration of active molecules
`is especially relevant to products designed to protect skin
`from photoaging. In this case long UVA rays penetrate deep
`into the epidermis and produce free radicals which can cause
`long term health effects. Antioxidants are able protect the
`cells from this damage by scavenging free radicals and inhib
`iting oxidation reactions. However, research has shown that
`in order formany of active molecules, such as antioxidants to
`be effective they must also reside in the epidermis.
`The most common technique to increase transdermal
`delivery is to use penetration enhancers. While many chemi
`cal penetration enhancers such as solvents work well to dis
`rupt the lamellar lipid structure of the skin many of them are
`toxic, irritating, allergenic, or not suited to cosmetic formu
`lations which cover large areas (unlike typical pharmaceuti
`cal transdermal patches).
`Other techniques to increase transdermal delivery of active
`compounds rely on delivery systems such as liposomes in
`combination with solvents. U.S. 2006/0110439 discloses a
`delivery system containing liposomes and solvents to
`increase penetration of an active compound. U.S. Pat. No.
`6,355,657 discloses a system for percutaneous delivery of the
`opioid loperamide that combines an organic solvent and fatty
`acid/fatty alcohol penetration enhancers, such as oleic acid,
`olleyl alcohol, ethoxydiglycol, laurocapram, alkanecarboxy
`lic acid, dimethyl sulfoxide, polar lipids or n-methyl-2pyr
`rolidone.
`While many cosmetic applications require the addition of
`penetration enhancers for efficacious actives, the enhancers
`must overcome the added challenges and needs of being safe,
`reversible and able to work in a cosmetic form or dose. In
`addition these enhancers must not interfere with the active
`molecule in such a way that compromises the molecule’s
`activity. Thus, there is a need for compositions that increase
`the transdermal bioavailability of active compounds.
`
`15
`
`20
`
`25
`
`30
`
`35
`
`40
`
`45
`
`50
`
`55
`
`BRIEF SUMMARY OF THE INVENTION
`
`60
`
`The invention provides an aqueous composition compris
`ing a) at least one active compound and b) at least one hydro
`trope in an amount effective to increase transdermal penetra
`tion of the active compound. Preferably, the hydrotrope is a
`cosmetically acceptable hydrotrope, such as nicotinamide,
`65
`caffeine, sodium PCA, or sodium salicylate. The active com
`pound can be poorly water soluble, requiring the need to be
`
`2
`solubilized by the hydrotrope itself (or a combination of
`hydrotropes), or a hydrophilic molecule that is readily dis
`solved.
`Another aspect of the invention provides a method for
`increasing the bioavailability of an active molecule compris
`ing applying an aqueous composition to skin, the composi
`tion comprisinga) at least one active compound and b) at least
`one hydrotrope in an amount effective to increase transdermal
`penetration of the active compound.
`A further aspect of the invention provides a method of
`preparing an aqueous composition comprising including in
`the composition at least one active compound and at least one
`hydrotrope in an amount effective to increase transdermal
`penetration of the active compound.
`These and other aspects of the invention are shown in the
`appended claims, and described in greater detail in the
`detailed description of the invention.
`
`BRIEF DESCRIPTION OF THE DRAWINGS
`
`FIG. 1 shows a diagrammatic representation of the model
`lipid bilayer assay.
`FIG. 2A and FIG. 2B show a table of glucose release from
`lipids in the presence of hydrotropes, and graph of glucose
`release after the presence of hydrotropes.
`FIG. 3A and FIG. 3B show graphs of the depth of penetra
`tion of vitamin C and baicalin in human ex-vivo skin.
`
`DETAILED DESCRIPTION OF THE INVENTION
`
`The present invention provides aqueous compositions
`comprising a) at least one active molecule and b) at least one
`hydrotrope in an amount effective to increase transdermal
`penetration of the active molecule.
`Applicants have discovered that hydrotropes in cosmetic
`formulations enhance bioavailability of active molecules.
`The function of increased bioavailability is due to the ability
`of the hydrotrope to increase depth of penetration through the
`skin. Hydrotropes can be used in all cosmetic formulas which
`contain more than 5% water, for both topical application and
`injection.
`Hydrotropes suitable for use in cosmetic compositions
`safely and reversibly disrupt the lamellar lipid crystalline
`layer of the stratum corneum in order to effectively deliver
`and increase bioavailability of active compounds in cosmetic
`formulations. When combined with poorly soluble and
`poorly penetrating active compounds such as polyphenols,
`adenosine, sugars, and hydrophilic molecules, hydrotropes
`increase penetration of the active compound through the skin.
`As used herein, improving bioavailability of an active com
`pound or molecule refers to increasing the amount of the
`active compound in skin, in comparison to a composition
`which does not contain the hydrotrope or hydrotropes present
`in the claimed aqueous compositions.
`As used herein, increasing dermal penetration of an active
`compound or molecule refers to increasing the amount or
`depth of penetration, or both, of an active compound through
`skin, preferably human skin, in comparison with a composi
`tion which does not contain the hydrotrope or hydrotropes
`present in the claimed aqueous compositions, or does not
`contain such hydrotropes in amounts effective to increase
`transdermal penetration.
`Hydrotropes (or hydrotropic agents) are a diverse class of
`water-soluble compounds that characterized by an
`amphiphilic molecular structure and ability to dramatically
`increase the solubility of poorly soluble organic molecules in
`water. Most hydrotropes have aromatic structure with anionic
`
`

`

`Case 1:17-cv-00868-JFB-SRF Document 13-7 Filed 08/18/17 Page 8 of 11 PageID #: 275
`
`US 9,018,177 B2
`
`4
`At least one hydrotrope refers to one or a combination of
`two or more hydrotropes. One or combination of two or more
`hydrotropes can be used to improve the bioavailability of
`active compounds, or increase transdermal penetration of
`active compounds.
`An advantage of using hydrotropes is, once a stable solu
`tion is obtained, further dilution doesn’t influence the stability
`of the solution. This is very different from organic solvents
`that are commonly used to increase the water solubility of
`actives. Typically, an aqueous dilution of organic solvents
`with pre-dissolved actives results in crystallization or precipi
`tation.
`Hydrotropes which function to increase the penetration of
`active molecules in the skin can be selected using models of
`the skin, such as the model lipid bilayer assay disclosed
`herein the Examples. An effective hydrotrope solution allows
`the reversible disruption of lipids, taking into consideration
`the dose and potential combination of cosmetic hydrotropes.
`The amount of hydrotrope present in the aqueous compo
`sitions will vary depending on the hydrotrope and the type
`and amount of active compound, preferably in the range of
`0.01% to 20%, with respect to the total weight of the compo
`sition.
`The inventive aqueous compositions comprise at least one
`active compound from various classes of compounds, such as
`compounds that are poorly water soluble (solubility.<0.1%),
`such as polyphenols, and compounds that are hydrophilic,
`such as vitamin C.
`The amount of active compound in the aqueous composi
`tion will depend on the identity of the active compound and its
`solubility in water, and the type and amount of hydrotrope
`present in the aqueous composition. Preferably, the amount
`ranges from 0.01% to 20%, based on the total weight of the
`composition.
`Polyphenols are a structural class of natural, synthetic, and
`semisynthetic organic compounds that have multiple phe
`nolic constituents. Polyphenols are normally available in
`plants and are very helpful to protect plants and also animals
`from usual health disorders and also the impacts of aging.
`Polyphenols function as potent free radical scavengers by
`donating their alcoholic hydrogen or one of their delocalized
`electrons. The two classes of polyphenols are flavonoids and
`non-flavonoids.
`Flavonoids are a specific group of polyphenols, and are
`further categorized, according to chemical structure, into
`chalcones, flavones, flavanones, flavanols, flavonols, dihy
`droflavonols, isoflavonoids, neoflavonoids, catechins, antho
`cyanidins, and tannins. Over 4,000 flavonoids have been
`identified, many of which occur in fruits, vegetables and
`beverages (tea, coffee, beer, wine and fruit drinks). The fla
`vonoids have been reported to have antiviral, anti-allergic,
`antiplatelet, anti-inflammatory, antitumor and antioxidant
`activities. Flavonoids protect lipids and vital cell components
`from damaging oxidative stress by efficiently scavenging free
`radicals.
`Non-flavonoid polyphenols include lignans, aurones, stil
`benoids, curcuminoids and other phenylpropanoids. Many of
`them are also well-known antioxidants like resveratrol, feru
`lic acid, curcumin, and pinoresinol.
`At least one polyphenol is solubilized in the aqueous com
`positions, and the amount of polyphenol will depend on the
`specific polyphenol and the type and amount of hydrotrope
`present in the aqueous compositions. The amount of polyphe
`nol present in the aqueous compositions can range from
`0.01% to 20%; about 0.1% to about 10%; or about 0.1% to
`about 5%, based on the total weight of the composition.
`
`3
`moiety, while some of them are linear alkyl chains, as listed in
`the table below. Although hydrotropes noticeably resemble
`surfactants and have the ability to reduce surface tension,
`their small hydrophobic units and relatively shorter alkyl
`chain distinguish them as a separate class of amphiphiles.
`Consequently their hydrophobicity is not sufficient enough to
`create well organized self-associated structures, such as
`micelles, even with a high concentration.
`Common hydrotropic molecules include: sodium 1,3-ben
`Zenedisulfonate, sodium benzoate, sodium 4-pyridinecar
`boxylate, sodium salicylate, sodium benzene sulfonate, caf
`feine, sodium p-toluene sulfonate, sodium butyl
`monoglycolsulfate, 4-aminobenzoic acid HCl, sodium
`cumene sulfonate, N,N-diethylnicotinamide, N-picolylnico
`tinamide, N-allylnicotinamide, 2-methacryloyloxyethyl
`phosphorylcholine, resorcinol, butylurea, pyrogallol, N-pico
`lylacetamide 3.5, procaine HC1, proline HCl, nicotinamide,
`pyridine, 3-picolylamine, sodium ibuprofen, sodium xylene
`sulfonate, ethyl carbamate, pyridoxal hydrochloride, sodium
`benzoate, 2-pyrrolidone, ethylurea, N,N-dimethylacetamide,
`N-methylacetamide, and isoniazid. Hydrotropes can be found
`in Lee J. et al., “Hydrotropic Solubilization of Paclitaxel:
`Analysis of Chemical Structures for Hydrotropic Property”,
`Pharmaceutical Research, Vol. 20, No. 7, 2003; and Hodgon
`T. K., Kaler E. W., “Hydrotropic Solutions”, Current Opinion
`in Colloid and Interface Science, 12, 121-128, 2007.
`Cosmetically acceptable hydrotropes refers hydrotropes
`that can be used in cosmetic compositions. While hydrotropes
`represent a broad class of molecules used in various fields,
`cosmetic applications will be limited due to safety and toler
`ance restrictions. Preferred hydrotropes in cosmetics are
`listed as below:
`
`Name of hydrotropes
`
`Nicotinamide (Vit B3)
`
`Structure
`
`O
`
`Caffeine
`
`Sodium PCA
`
`| S NH2
`2
`º
`N sº
`& I Ä
`^ CH3
`
`CH3
`
`O
`
`H O
`
`O
`
`N
`
`ONa.
`
`Sodium Salicylate
`
`O.
`
`O
`
`Na"
`
`OH
`
`10
`
`15
`
`20
`
`25
`
`30
`
`35
`
`40
`
`45
`
`50
`
`55
`
`60
`
`The suitability of a hydrotrope for use in cosmetic compo
`sitions can be determined using tests known in the art for
`determining effects of compounds on skin, and bioavailabil
`ity methods.
`
`65
`
`

`

`Case 1:17-cv-00868-JFB-SRF Document 13-7 Filed 08/18/17 Page 9 of 11 PageID #: 276
`
`US 9,018,177 B2
`
`6
`For injection, the composition can be provided in the form
`of aqueous or oily lotions or in the form of serums. For the
`eyes, the composition can be provided in the form of drops
`and, for ingestion, it can be provided in the form of capsules,
`granules, syrups or tablets.
`The amounts of the various constituents of the composi
`tions according to the invention are those conventionally used
`in the fields under consideration.
`In the cosmetics field, these compositions constitute in
`particular creams for cleaning, protecting, treating or caring
`for the face, for the hands, for the feet, for the major anatomi
`cal folds or for the body (for example, day creams, night
`creams, anti-aging creams, moisturizing creams, make-up
`removing creams, foundation creams or sun creams), liquid
`foundations, make-up-removing milks, protective or care
`body milks, sun milks, lotions, gels or foams for caring for the
`skin, such as cleansing lotions, sun lotions, artificial tanning
`lotions, bath compositions, deodorizing compositions com
`prising a bactericidal agent, aftershave gels or lotions, depila
`tory creams, compositions for countering insect stings or
`bites, pain-relieving compositions or compositions for treat
`ing certain skin diseases, such as eczema, rosacea, psoriasis,
`lichen and severe pruritus.
`Another aspect of the invention provides a method for
`preparing the aqueous compositions comprising including in
`the composition at least one active compound and at least one
`hydrotrope in an amount effective to increase transdermal
`penetration of the active compound. A hydrotrope solution is
`prepared by completely dissolving one or more hydrotropic
`agents into water, in an amount effective to increase transder
`mal penetration of the active compound. If the active com
`pound is poorly soluble in water, the amount of hydrotrope
`should also be effective to solubilize the active compound.
`The active compound, such as a polyphenol, is then added in
`and mixed using stirring bar or any other mixer. A clear stable
`solution with a concentration that does not exceed the solu
`bility of the active compound would be ready after more than
`one hour of mixing. No heat is necessary by following this
`procedure to dissolve polyphenols. Everything is prepared at
`room temperature to keep the stability of polyphenols. This is
`extremely useful to protect the activity of certain compounds
`and also makes the process much easier.
`
`EXAMPLES
`
`Example 1
`
`Model Lipid Bilayer Assay
`
`In this model system the lipid bilayers are able entrap a
`detectable compound and upon lipid disorder (i.e. in the pres
`ence of a hydrotrope) the compound can be released from the
`structure and measured in a chemical assay.
`A representative lipid mixture (POLYGLYCERYL-3
`CETYL ETHER (47.5%) (and) CHOLESTEROL (47.5%
`(and) DICETYL PHOSPHATE.5%) was selected for the lipid
`vesicles and glucose was chosen as the entrapped compound.
`Glucose release can be measured by glucose-oxidase assay
`using a Sigma HK assay kit. Higher glucose release in the
`presence of a hydrotrope suggests that that particular hydro
`trope could have a greater effect in penetration improvement.
`The experimental setup is as follows: 1% lipids were added
`to a 0.3M glucose solution and stirred for 6 hours (rpm 1100)
`at 85°C. Final lipid solution was dialysized for 24 hrs in 0.7%
`NaCl bath using a 3.5 Kd dialysis membrane in order to
`remove any free glucose in the supernatant. To ensure the
`remaining glucose is fully entrapped in the lipid vesicle the
`
`5
`Increasing the water solubility of polyphenols refers to
`increasing the solubility of the polyphenol in water in com
`parison with solubility of the polyphenol in water in the
`absence of the hydrotrope or hydrotropes.
`If the active compound is poorly soluble in water, the at
`least one hydrotrope is present in the aqueous composition in
`amounts effective to increase the solubility of the active com
`pound, such as a polyphenol, in water. The amount of hydro
`trope will vary depending on the hydrotrope and the type and
`amount of the active compound. The amount of hydrotrope
`present in the aqueous compositions can range from 0.1% to
`60%; about 0.1 to about 50%; or about 1% to 50%, based on
`the total weight of the composition.
`The aqueous compositions can also comprise at least one
`additive conventionally used in the cosmetics field and which
`does not affect the properties of the compositions according
`to the invention, such as thickeners, fragrances, pearlescent
`agents, preservatives, sunscreens, anionic or nonionic or cat
`ionic or amphoteric polymers, proteins, protein hydrolysates,
`fatty acids, such as 18-methyleicosanoic acid, vitamins, pan
`thenol, silicones, vegetable, animal, mineral or synthetic oils,
`gelling agents, antioxidants, solvents, fillers, screening
`agents, odor absorbers and coloring materials. These addi
`tives can be present in the composition according to the inven
`tion in proportions which are not limited, but which prefer
`ably fall in the range from 0 to 50% by weight, 5-40% by
`weight, or 30-50% by weight with respect to the total weight
`of the composition.
`The aqueous compositions can comprise at least one active
`compound and at least one hydrotrope in an amount effective
`to increase transdermal penetration of the active compound,
`with water making up the remainder of the composition.
`Preferably, the composition comprises from 5% to 99.5% by
`weight of water, with respect to the total weight of the com
`35
`position.
`The pH of the aqueous compositions is not limited but is
`generally between 2 and 12 and preferably between 3 and 9.
`The pH can be adjusted to the desired value by addition of a
`base (organic or inorganic) to the composition, for example
`ammonia, sodium hydroxide, potassium hydroxide, or a pri
`mary, secondary or tertiary (poly)amine, such as monoetha
`nolamine, diethanolamine, triethanolamine, isopropanola
`mine or 1,3-propanediamine, or with basic amino acids or
`poly amino acids like lysine, or arginine, or alternatively by
`addition of an inorganic or organic acid, preferably a carboxy
`lic acid, such as, for example, citric acid.
`Generally, any composition of the invention can be
`ingested, injected or topically applied to the skin (over any
`cutaneous region of the body) or to the mucous membranes
`(oral, jugal, gingival, genital, conjunctival, and the like).
`Preferably, the compositions of the invention are topically
`applied onto the skin or mucous membranes. Depending on
`the method of administration under consideration, the com
`position can be provided in any dosage form normally used.
`55
`For topical application to the skin, the composition can
`have the form in particular of aqueous or oily solutions or of
`dispersions of the lotion or serum type, of emulsions with a
`liquid or semi-liquid consistency of the milk type, obtained
`by dispersion of a fatty phase in an aqueous phase (O/W) or
`vice versa (W/O), or of suspensions or emulsions with a soft
`consistency of the aqueous or anhydrous gel or cream type, or
`else of microcapsules or microparticles, or of vesicular dis
`persions of ionic and/or nonionic type or of foams. In fact any
`composition having between 5 to 99.5% of water phase is
`relevant. These compositions are prepared according to the
`usual methods.
`
`10
`
`15
`
`20
`
`25
`
`30
`
`40
`
`45
`
`50
`
`60
`
`65
`
`

`

`Case 1:17-cv-00868-JFB-SRF Document 13-7 Filed 08/18/17 Page 10 of 11 PageID #: 277
`
`US 9,018,177 B2
`
`7
`supernatant is measured by the glucose oxidase kit. As a
`control the lipid vesicles can be broken by n-octylglucoside
`and the entrapped glucose can be measured.
`The results shown in FIG. 2 represent a lipid suspension
`that was incubated with hydrotropes for 20 hours. The higher
`release of glucose suggests a more efficient hydrotrope or
`combination of hydrotropes. The following hydrotropes were
`tested both for dose response and potential synergy when
`combined with each other: caffeine, niacinamide, NaPCA,
`and sodium salicylate.
`Hydrotropes or combinations of hydrotropes with >15%
`glucose release were selected as preferred hydrotropes for
`formulations.
`
`Example 2
`
`Ex-Vivo Testing of Hydrotropes as Penetration
`Enhancers
`
`-continued
`
`PHASE
`
`INCI US Name
`
`CONCENTRATION (wt %)
`
`C
`PH
`
`BAICALIN
`SODIUM HYDROXIDE
`
`0.1
`O
`
`The compounds of phase A were combined and heated to
`~65-75° C. under constant stirring. Phase A was then cooled
`and phase B was added. Separately, the compounds of phase
`C were combined and mixed under constant stirring at room
`temperature. Phase A+B was added to phase C and stirred for
`one hour to form the serum. The pH of the serum was adjusted
`to 4.5 with sodium hydroxide.
`3.2 Oil in Water Emulsion
`The oil in water emulsion was prepared using the following
`method:
`
`10
`
`15
`
`20
`
`25
`
`30
`
`To further evaluate the effect of penetration enhancers on
`human skin we performed CARS (coherent anti-stokes raman
`spectroscopy) experiments. Using this technique we are able
`to follow the penetration depth of antioxidants in human
`ex-vivo skin and compare the variability between a formula
`tion containing no hydtrotropes, one hydrotrope, and combi
`nation of hydrotropes.
`The results in FIG. 3 show the penetration depth of the
`antioxidants vitamin C and baicalin (solubilised by niacina
`mide and caffeine) in ex-vivo human skin after 2, 6, and 16
`hours with the following formulas–Formula A (glycol
`serum with 10% vitamin C), Formula B (glycol serum with
`10% vitamin C, 0.5% baicalin, and 5% caffeine), and For
`mula C (glycol serum with 10% vitamin C, 0.5% baicalin, 5%
`caffeine and 5% niacinamide). Formula A does not contain
`baicalin as it requires the addition of some hydrotropes for
`aqueous solubility. In FIG. 3A, Formula A is represented by
`the left column; Formula B by the middle column; and For
`mula C by the right column. In FIG. 3B, Formula B is repre
`sented by the left column; and Formula C by the right column.
`The results clearly show that the addition of hydrotropes to
`formulations increases the penetration depth of the antioxi
`dant molecules. In this case the synergy of niacinamide and
`caffeine allow for the highest penetration depth of both vita
`min C and baicalin.
`Of interesting note, baicalin requires the association of
`45
`hydrotropes in order to achieve efficient levels in aqueous
`environments while vitamin C is highly hydrophilic and does
`not. However, the addition of hydrotropes in the final formu
`lation improves the penetration depth of both molecules.
`Example 3
`
`35
`
`40
`
`50
`
`Compatible Formula Compositions
`
`3.1 Serum
`The serum was prepared using the following method:
`
`PHASE INCI US Name
`
`CONCENTRATION (wt %)
`
`A.
`A.
`A.
`B
`C
`C
`C
`C
`
`DIPROPYLENE GLYCOL
`PEG-4
`BAICALIN
`ALCOHOL DENATURED
`WATER
`ASCORBIC ACID
`CAFFEINE
`NLACINAMIDE
`
`10
`10
`0.4
`10
`49.5
`10
`5
`5
`
`55
`
`60
`
`65
`
`PHASE INCI US Name
`
`Concentration
`(wt %)
`
`A.
`A.
`A.
`A.
`A.
`A.
`A.
`A.
`B
`B
`B
`
`B
`B
`B
`B
`B
`B
`
`WATER
`NIACINAMIDE
`CAFFEINE
`BAICALIN
`POLYSORBATE 61
`PHENOXYETHANOL
`SODIUM STEAROYL GLUTAMATE
`DISODIUM EDTA
`OCTOCRYLENE
`ISOPROPYL LAUROYL SARCOSINATE
`OCTYLDODECANOL (and)
`OCTYLDODECYL XYLOSIDE
`GLYCERYL STEARATE
`STEARYL ALCOHOL
`CAPRYLYL GLYCOL
`p-ANISIC ACID
`ISOTRIDECYL ISONONANOATE
`BASF W RP
`
`46.78
`2.7
`2.7
`0.27
`0.36
`0.5
`0.4
`0.1
`2.5
`5
`0.86
`
`0.29
`0.14
`0.3
`0.1
`32
`5
`
`The compounds of phase A were combined and heated to
`80-85°C. mixing slowly (500-700 rpm) until all dissolved.
`Separately, the compounds of phase B were combined and
`heated to ~80-85°C. mixing slowly (500-700 rpm) until all
`dissolved. Phase B was added to phase under mixing condi
`tions (Rayneri). The mixing speed was increased and the
`mixture was emulsified for thirty minutes. Particle size was
`then checked under a microscope. The resultant oil in water
`emulsion was cooled to 25°C. with slower speed mixing. The
`emulsion was then run under a high pressure homogenizer.
`
`What is claimed is:
`1. An aqueous cosmetic skin-care composition compris
`ing:
`(a) about 0.5 wt. 9% of baicalin and/or about 10 wt. 9%
`vitamin C:
`(b) about 5 wt.% of each of the hydrotropes caffeine and
`niacinamide, wherein the amount of said hydrotropes is
`effective to increase transdermal penetration of baicalin
`and/or vitamin C; and
`(c) water;
`wherein said skin care composition is in the form of a
`lotion, serum, gel, milk, foam, liquid foundation, or
`cream, and is suitable for topical application to the skin.
`2. The composition of claim 1, wherein the composition is
`a Serul III.
`3. The composition of claim 2, wherein the serum further
`comprises dipropylene gly