mm.
`
`,======é.2..=======’
`H0\m0\¢o
`
`
`
`w 56/01 ”If
`
`:LL L'f/
`
`
`
`ZOF<O_n:ww<n_Omew_
`
`PROVISIONAL
`APPLICATION
`NUMBER
`
`Form PTO-1625
`(Rev. 6/99)
`
`Amneal 1031
`
`(
`
`FACE)
`
`Amneal v. Supernus
`|PR2013-00368, 2013-00371 and
`2013-00372
`
`
`
`
`1
`
`
`
`
`

`

`PATENT AEPLICATION
`HHHHUUHMWNW/l
`
`
`
`
`
`5' APPBGVED FOR LICENSE
`fiNIT‘IALS
`
`
`
`.. "
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`(LEFT OUTSIDE)
`2
`
`2
`
`

`

`
`
`PATENT APPLICATION SERIAL NO.
`
`US. DEPARTMENT OF COMMERCE
`
`PATENT AND TRADEMARK OFFICE
`FEE RECORD SHEET.
`
`04/10/8001 UTRUDNBI 00000001 60201918
`
`01 1713:2113
`
`75.00 DP
`
`PTO-1556
`
`(5/87)
`'U.S. GPO: 2000-468-987/39595
`
`3
`
`

`

`Page l-of 1
`
`
`
`
`UNITED STATES PATENT AND ThADEMARK OFFICE
`COMMISSIONER FOR PATENTS
`UNITED STATES PATENT AND TRADEMARK OFFICE
`MSHINGTDN, D.C. 2023i
`www.uspio.gov
`
`lllllllllllllllllllllllllllllllllllllllllllllllllllll
`Bib Data Sheet
`
`CONFIRMATION NO. 4705
`
`SERIAL NUMBER
`60/281,916
`
`A PPLICANTS
`
`FILING DATE
`04/05/2001
`RULE
`
`‘
`
`CLASS
`
`GROUP ART UNIT
`
`ATTORNEY
`DOCKET NO.
`512-53 P
`
`Robert A. 'Ashley, Newton, PA;
`
`. * CON TIN U ING I DATA it************************
`
`, * FOREIGN APPLICATIONS ********************
`
`IF REQUIRED, FOREIGN FILING LICENSE
`GRANTED ** 06/16/2001
`'
`
`** SMALL ENTITY **
`
`Foreign Priority claimed
`35 use 119 (a-d) condltlons
`met
`
`D yes D no
`. [:1 yes [I no El Met afler
`Allowance
`
`Examiner's Si- nature
`
`Initials
`
`STATE OR
`SHEETS ,
`COUNTRY DRAWING
`
`TOTAL
`CLAIMS
`
`INDEPENDEN
`CLAIMS
`
`for following: ..
`
`CI All Fees
`
`Cl 1.16 Fees (Filing)
`
`D 1 17 Fees ( Processing Ext of
`time)
`
`U1.18Fees(rssue)
`._-__
`
`.m_
`
`CI Credit
`
`FILING FEE FEES: Authority has been given in Paper
`RECEIVED No.
`to charge/credit DEPOSIT ACCOUNT
`
`4
`
`

`

`
`
`3,; |_ndividualName
`
`_ddress
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`Li Form
`
`AIPrOI/
`
`'
`'
`'
`Please type a plus sIgn M mSIde thb pox
`
`—>
`
`PTO/SB/16 a00
`—_" "'
`Approved for use through10/31/2002. OMB 0655-0033
`QE 2;
`U.8. Patent and Trademark Office; US. DEPARTMENT OF COMMERCE
`as,E u
`, B a‘ I-Under the Paperwork Reduction Act of 1995 no persons are required to respond to a collection of information unless it displays a valid OMB control number.
`01a I:
`BE 9,
`PROVISIONAL APPLICA TION FOR PA TENT COVER SHEET
`lee—2% ’
`This is a request for filing a PROVISIONAL APPLICATION FOR PATENT under 37 CFR .1'53(°)'
`
`a ,
`
`Given Name (first and middle [if any])
`
`INVENTOR S
`Family Name or Surname
`
`Residence
`Cit and either State or Forei-n Count
`
`Robert A.
`
`Ashley
`
`
`
`63 Woodhill Road, Newtown, PA 18940
`
`,
`
`E
`&
`fig ,1.
`%\Iact .
`E—‘—o=5
`
`Additional inventors are being named on thI___ separately numbered sheets attached hereto
`
`TITLE OF THE INVENTION (280 characters max)
`
`METHODS OF TREATING ACNE
`
`Direct all correspondence to:
`CORRESPONDENCE ADDRESS
`D E:l —»
`
`Bar Code Label here
`
`Type Customer Number here
`
`we
`t
`
`Firm or
`
`
`
`HOFFMANN 8. BARON, L.L.P.
`
`—I_ Teiepnone (516)822-3550 2-(516)822-3582
`
`ENCLOSED APPLICATION PARTS check all that a . -I
`
`g Specification Number of Pages
`V Drawin s N b
`rsn t
`
`- D CD(s) Number
`
`|::l
`
`E] Application Data Sheet. see 37 CFR 1.76
`METHOD OF PAYMENT OF FILING FEES FOR THIS PROVISIONAL APPLICATION FOR PATENT (check one)
`
`VA ‘
`Applicant claims small entity status. See 37 CFR 1.27.
`DE
`A check or money order is enclosed to cover the filing fees
`D The Commissioner is hereby authorized to charge filing J:|
`fees or credit any overpayment to Deposit Account Numbe
`D Payment by credit card. Form PTO-2038 is attached.
`The invention was made by an agency of the United States Government or under a contract with an agency of the
`United States Government.
`D No.
`D Yes, the name of the US. Government agency and the Government contract number are:
`
`' FILING FEE
`AMOUNT ’
`
`$75-00
`
`
`
`Respectfully submitted,
`
`enemies AA A Aer
`
`Date
`
`4/5/2001
`
`mam.
`
`,
`
`TYPED or PRINTED NAME Susan A- SIPOS. Esti-
`
`0" aPP’OP”a’9)
`
`‘
`
`
`
`(516) 608-4710
`TELEPHONE
`USE ONLY FOR FILING A PROVISIONAL APPLICATION FOR PA TENT
`This collection of information is required by 37 CFR 1. 51 The information is used by the public to file (and by the PTO to process) a provisional
`application. ConfidentialityIs governed byy35 U.3.0.122 and 37 CFR 1.14. This collectIon Is estimated to take 8 hours to complete, including
`gathering, preparing, and submitting the complete provisional application to the PTO Time will vary depending upon the individual case. Any
`comments on the amount of time you require to complete this form and/or suggestions for reducing this burden should be sent to the Chief
`Information Officer, U. S. Patent and Trademark Office, U. S. Department of Commerce Washin ton, D. C. 20231. DO NOT SEND FEES OR
`COMPLETED FORMS TO THIS ADDRESS. SEND TO: Box Provisional Application, Assistant
`ommissioner for Patents, Washington D.C.
`
`nAr
`'r\ u' n
`P198MALLIREV05
`m, Acidswol wuoflflms
`I hereby certify that on the date indicated above, I deposited this paper or fee
`with the United States Posta| Service and that it was addressed for delivery
`to Assistant Commissioner for Patents, Washington, DC. 2023 I by
`"EXPRESSMAILPostOfficetoA
`see" Sew.
`
`5 3
`.Is.
`(Print Name)
`(SignaGHE)
`
`
`5
`
`

`

`
`
`PROVISIONAL APPLICA TION COVER SHEET
`Additional Page
`
`
`
`
`
`PTO/SB/16 (8-00)
`Approved for use throu h 10/31/2002. OMB 0651-0032
`U.S. Patent and Trademark Office; US. D PARTMENT OF COMMERCE
`Under the Paperwork Reduction Act of 1995, no persons are required to resend to a c ollection of information unless it dis la 5 a valid OMB control number.
`
`'
`Type a plus sign (+)
`
`
`
`INVENTOR(§)LAPPLICANT(S)
`
`
`
`
`
`Residence
`
`
`Cit and either State or Forei- n Count
`Given Name_(flrst and middle [if anyl)
`Familvgr Surname
`
`
`
`
`
`
`Number
`
`2
`
`of
`
`2
`
`WARNING: Information on this form may become public. Credit card information should not
`be included on this form. Provide credit card information and authorization on PTO-2038.
`
`6
`
`

`

`METHODS OF TREATING ACNE
`
`BACKGROUND OF THE INVENTION
`
`Acne is a common inflammatory disease in skin areas where sebaceous glands are ‘
`
`largest, most numerous, and most active. In its mildest form, acne is a more or less
`
`5
`
`superficial disorder characterized by slight, spOtty skin irritations. In such cases, ordinary
`
`skin hygiene is typically a satisfactory treatment. In the more inflammatory types of acne,
`
`however, pustules; infected cysts; and in extreme cases, canalizing, inflamed and infected
`
`sacs appear. Without effective treatment, these lesions may become extensive and leave
`
`permanent, disfiguring scars.
`
`1 0
`
`15
`
`20
`
`25
`
`Micro-organisms, especially Propionibacterium acnes, are strongly implicated in
`
`the pathogenesis of acne. The micro-organisms are thought to release microbial mediators
`
`of inflammation into the dermis or trigger the release of cytokines from ductal
`
`keratinocytes.
`
`Accordingly, the efficacy of antibiotics in treating acne is thought to be due, in
`
`significant part, to the direct inhibitory effect of the antibiotics on the growth and
`
`metabolism of these micro-organisms. Systemically-administered tetracycline antibiotics,
`
`.
`
`especially minocycline hydrochloride, are particularly effective in treating acne.
`
`The tetracyclines are a class of compounds of which tetracycline is the parent
`
`compound. Tetracycline has the following general structure:
`
`
`
`
`
`7
`
`

`

`
`
`Structure B
`
`
`
`The numbering system of the multiple ring nucleus is as follows:
`
`5
`
`Tetracycline, as well as the 5-OH (oxytetracycline, e. g. Terramycin) and 7—Cl
`
`(chlorotetracycline, e. g. Aureomycin) derivatives, exist in nature, and are all well known
`
`antibiotics. Semisynthetic derivatives such as 7-dimethylamino-tetracycline (minocycline)
`
`10
`
`and 6a—deoxy—5-hydroxy-tetracycline (doxycycline) are also known tetracycline
`
`antibiotics. Natural tetracyclines may be modified without losing their antibiotic
`
`properties, although certain elements of the structure must be retained to do so.
`
`i:
`it;
`
`In addition to the direct antimicrobial activity of tetracyclines, further activities of
`
`15
`
`antibiotic tetracyclines have been investigated for possible therapeutic effects on acne.
`
`For example, a study by Elewski et al., J. Amer. Acad; Dermatol. , 8:807-812
`
`(1983) suggests that acne therapy, consisting of orally-administered tetracycline at a total
`
`daily dose of 1000mg, may have therapeutic anti-inflammatory effects in addition to
`
`20
`
`antimicrobial effects. In particular, it was found that tetracycline inhibited neutrophil
`
`chemotaxis induced by bacterial chemotactic factors.
`
`However, a more recent study performed by Eady et al., J. Invest. Dermatol.,
`
`101:86-91 (1993) found somewhat different results with respect to the effect of
`
`25
`
`tetracyclines on cytokines. The study was designed to determine whether oral acne
`
`therapy with minocycline or tetracycline altered the cytokine content of open comedones.
`
`The total daily dose of minocycline administered was 100mg. The total daily dose of
`
`tetracycline administered was 1000mg. It was found that these therapies upregulated the
`
`production of bioactive IL-lu-like material and immunochemical IL-lB. IL-1 is
`
`30
`
`' considered to be a pro—inflammatory cytokine. The authors speculate that increased levels
`
`if:
`at;
`int.
`11.3“:
`
`E i
`
`i;
`
`ii
`iii
`iii
`a»;
`
`8
`
`

`

`
`
`_ ,\
`
`of IL-1 in comedones destined to become inflamed may enhance resolution and promote.
`
`repair of the damaged follicular epithelium.
`
`Another possible activity of tetracyclines in acne therapy was investigated by
`
`Bodokh, I., et al., Acta. Derm. Venerol., 77:255-259 (1997). Their study was designed to
`
`evaluate the action of minocycline on sebaceous excretion in acne patients. A 100mg daily 7
`
`dose of minocycline was administered. A subclinical increase in seborrhoea was reported.
`The authors propose that minocycline induces an increase in seborrhoea via a reduction in
`
`ductal obstruction. The mechanism by which the ductal obstruction is reduced is proposed
`
`to be the reduction in ductal irritation. The authors suggest that the reduction of ductal
`
`irritation is due to minocycline’s direct effect on P. acnes, or minocycline’s effect on the
`
`lipase produced by P. acnes.
`
`Bodokh et al. also found that during treatment no correlation exists between
`
`seborrhoea intensity and clinical severity of acne. The authors state that the lack of
`
`correlation shows that seborrhoea is pathogenic because it is the “culture medium” of P.
`
`acnes. Thus, it can be concluded that the authors consider the antimicrobial activity of
`minocycline as a significant therapeutic activity with respect to acne.
`
`Similarly, in a recent clinical study it was found that tetracycline doses lower than.
`antimicrobial doses had no clinical effect on acne. (Cunliffe et al., J. Am. Acad.
`Dermatol, 16:591-9 (1987).) In particular, a 100mg total daily dose of minocycline; and a
`
`1.0g total daily dose of tetracycline were found to be necessary to successfully treat acne.
`
`The antimicrobial effects of antibiotics are generally directly proportional to the
`
`dose administered of the antibiotics. Accordingly, in moderate to severe (i.e.
`
`inflammatory) forms of acne, oral antibiotics are typically administered at high doses. For
`
`example, in conventional acne therapy, tetracycline is administered at an initial dose of
`
`500 to 2,000 mg/day, followed by a maintenance dose of 250-500 mg/day.
`
`553*mu.
`
`10
`
`15
`
`20
`
`25
`
`30
`
`
`
`9
`
`

`

`
`
`Clearly, the state—of—the-art teaching is that the clinical efficacy of systemically—
`
`administered tetracyclines in the treatment of acne is due in significant part to the
`
`antimicrobial effects of the tetracyclines.
`
`5
`
`The use of tetracycline antibiotics, however, can lead to undesirable side effects.
`
`For example, the long term administration of antibiotic tetracyclines can reduce or
`
`eliminate healthy microbial flora, such as intestinal flora, and can lead to the production of
`
`antibiotic resistant organisms or the overgrowth of yeast and fungi. Other side effects
`
`include gastrointestinal symptoms such as nausea, vomiting, abdominal pain, diarrhea,
`
`10
`
`rashes, and other allergic reactions. Tetracyclines also can cause fetal harm if used during
`
`pregnancy.
`
`.
`
`it“:
`£33
`ii;
`iii
`
`Accordingly, there is a need for an effective treatment of acne which does not
`
`cause the undesirable side effects produced by the systemically-administered antibiotics
`
`15
`
`used in conventional acne therapy. ‘
`
`SUMMARY OF INVENTION
`
`The present invention provides a method of treating acne in a human in need
`
`20
`
`thereof. The method comprises administering systemically to the human a tetracycline
`
`compound in an amount that is effective to treat acne but has substantially no
`
`antimicrobial activity, without administrating a bisphosphonate compound.
`
`25
`
`30
`
`BRIEF DESCRIPTION OF THE DRAWINGS
`
`Figure 1 shows the photoirritancy factor (PIF) for some tetracycline compounds.
`
`For structure K, the compounds indicated are as follows:
`
`COL
`
`R7
`
`308
`31 l
`306
`
`hydrogen
`hydrogen
`hydrogen
`
`R8 .
`
`hydrogen
`hydrogen
`hydrogen
`
`4
`
`R9
`
`amino
`palmitamide
`dimethylamino
`
`
`
`if?
`is;
`{3%
`
`a 1
`
`333
`11m"i:
`m,
`3:“;ii
`3.3 §
`ri:
`in:
`
`10
`
`

`

`
`
`For structures L, M, N or O the compounds indicated are as follows:
`
`COL
`
`‘
`
`. 5
`
`801
`802
`804
`805
`
`10
`
`R7
`
`R8
`
`R9
`
`hydrogen
`hydrogen
`hydrogen
`hydrogen
`
`hydrogen
`hydrogen
`hydrogen
`hydrogen
`
`acetamido
`dimethylaminoacetamido
`nitro ‘
`amino
`
`For structure P, R7 is hydrogen, R8 is hydrogen and R9 is nitro.
`
`DETAILED DESCRIPTION
`
`15
`
`The present invention provides methods of treating acne. As used herein, the term
`
`\ ”acne" is a disorder of the skin characterized by papules, pustules, cysts, nodules,
`
`comedones, and other blemishes or skin lesions. These blemishes and lesions are often
`
`accompanied by inflammation of the skin glands and pilosebaceous follicles, as well as,
`
`microbial, especially bacterial, infection.
`
`20
`
`For the purposes of this specification, acne includes all known types of acne Some
`
`types of acne include, for example, acne vulgaris, cystic acne, acne atrophica, bromide
`
`acne, chlorine acne, acne conglobata, acne cosmetica, acne detergicans, epidemic acne,
`
`acne estivalis, acne fialminans, halogen acne, acne indurata, iodide acne, acne keloid, acne
`
`25
`
`mechanica, acne papulosa, pomade acne, premenstral acne, acne pustulosa, acne rosacea,
`
`acne scorbutica, acne scrofulosorum, acne urticata, acne varioliformis, acne venenata,
`
`propionic acne, acne excoriee, gram negative acne, steroid acne, and nodulocystic acne.
`
`The present invention can also be used to treat certain other types of acneiform
`
`30
`
`dermal disorders, e. g. perioral dermatitis, seborrheic dermatitis in the presence of acne,
`
`gram negative folliculitis, sebaceous gland dysfunction, hiddradenitis suppurativa, pseudo-
`
`folliculitis barbae, or folliculitis.
`
`11
`
`11
`
`

`

`
`
`The method comprises the administration of a tetracycline compound to a human
`in an amount which is effective to treat acne, but which has substantially no antimicrobial
`activity.
`
`5
`
`The tetracycline compound can be an antimicrobial or non-antimicrobial
`compound. The tetracycline compound has the general structure indicated above.
`
`13?
`£33
`g“;
`i”;
`if?
`’ :33
`
`$3
`
`ES:
`
`€33
`
`10
`
`15
`
`20
`
`25
`
`Some examples ofantimicrobial tetracycline compounds include doxycycline,
`minocycline, tetracycline, oxytetracycline, chlortetracycline, demeclocycline, lymecyclinc
`and their pharmaceutically acceptable salts. Doxyclycline is preferably administered as its
`hyclate salt or as a hydrate, preferably monohydrate.
`
`Non-antimicrobial tetracycline compounds are structurally related to the antibiotic
`tetracyclines, but have had their antibiotic activity substantially or completely eliminated
`by chemical modification. For example, non-antimicrobial tetracycline compounds are
`capable ofachieving antibiotic activity comparable to that oftetracycline at concentrations
`at least about ten times, preferably at least about twenty five times, greater than that of
`tetracycline.
`
`Examples ofchemically modified non-antimicrobial tetracyclines (CMT’s)
`include, 4-de(dimethylamino)tetracycline (CMT—l), tetracyclinonitrile (GMT-2), 6-
`demethyl-6-deoxy-4-de(dimethylamino)tetracycline (GMT-3), 7-chloro-4-
`de(dimethylamino)tetracycline (GMT-4), tetracycline pyrazole (GMT-5), 4-hydroxy-4-
`de(dimethylamino)tetracycline (GMT-6), 4-de(dimethylamino-12a-deoxytetracycline
`(GMT-7), 6-deoxy-5a-hydroxy-4-de(dimethylamino)tetracycline (GMT-8), 4-
`de(dimethylamino)-12a-deoxyanhydrotetracycline (GMT-9), 4-
`de(dimethylaminojminocycline (GMT-10).
`
`Further examples of chemically modified non-antimicrobial tetracyclines include
`Structures C-Z. (See Index of Structures.)
`
`30
`
`12
`
`12
`
`

`

`The minimal amount of the tetracycline compound administered to a human is the
`
`lowest amount capable of providing effective treatment of acne. Effective treatment is a
`
`reduction or inhibition of the blemishes and lesions associated with acne.
`
`The amount of a tetracycline compound that has substantially no antimicrobial
`
`activity is an amount that does not significantly prevent the growth of microbes, e. g.
`
`bacteria.
`
`For example, tetracycline compounds that have significant antimicrobial activity
`
`5
`
`,
`
`10
`
`may be administered in an amount which is 10-80% of the antimicrobial amount. More
`
`preferably, the antimicrobial tetracycline compound is administered in an amount which is
`
`40-70% of the antimicrobial amount.
`
`Some examplesof antimicrobial amounts of members of the tetracycline family
`
`15
`
`include 100mg/day of doxycycline, 200mg/day of minocycline, 250mg of tetracycline four
`
`times a day, lOOOmg/day of oxytetracycline, 600mg/day of demeclocycline and
`
`600mg/day of lymecycline.
`
`An example of an antimicrobial tetracycline administered in a non-antimicrobial
`
`20
`
`amount is doxycycline hyclate administered at a 20 milligram dose twice daily. Such a
`
`formulation is sold for the treatment of periodontal disease by CollaGenex
`
`Pharmaceuticals, Inc. of Newtown, Pennsylvania under the trademark Periostat ®.
`
`The tetracycline compound may be administered by sustained release. Sustained
`
`25
`
`release administration is a method of drug delivery to achieve a certain level of the drug
`
`over a particular period of time. The level typically is measured by serum concentration.
`
`Further description of methods of delivering tetracycline compounds by sustained release
`
`can be found in the patent application, “Controlled Delivery of Tetracycline and
`
`Tetracycline Derivatives,” filed on April 5, 2001 and assigned to CollaGenex
`
`30
`
`Pharmaceuticals, Inc. of Newtown, Pennsylvania. The aforementioned application is
`
`incorporated herein by reference in its entirety.
`
`Cl“:
`$3}
`it;
`
`g!)
`5.3”}
`
`:Hr
`{HE
`rain:
`we
`
`ii?
`if?X ‘
`
`isn't:
`
`13
`
`
`
`13
`
`

`

`
`
`For example, 40 milligrams of doxycycline may be administered by sustained
`
`release over a 24 hour period.
`
`5
`
`The antimicrobial tetracycline compound can be administered in an amount which
`
`results in a serum tetracycline concentration which is 10-80% of the minimum
`
`antimicrobial serum concentration. The minimum antimicrobial serum concentration is
`
`the lowest concentration known to exert a significant antimicrobial effect.
`
`10
`
`Some examples of the approximate minimum antimicrobial serum concentrations
`
`of members of the tetracycline family follow.
`
`i393
`€23
`1‘13
`ii:
`ifixa‘.
`rift
`gut.
`£3”?
`3..
`£31':
`ab:
`
`1;
`3.5?
`’3”?
`ga
`
`A single dose of two 100mg minocycline HCl tablets administered to adult humans
`
`results in minocycline serum levels ranging from 0.74 to 4.45 ug/ml over a period of an
`
`15
`
`hour. The average level is 2.24 rig/ml.
`
`Two hundred and fifty milligrams of tetracycline HCl administered every six hours
`
`over a twenty-four hour period produces a peak plasma concentration of approximately 3
`
`ug/ml. Five hundred milligrams of tetracycline HCl administered every six hours over a
`
`20
`
`twenty-four hour period produces a serum concentration level of 4 to 5 ug/ml.
`
`The minimum antimicrobial serum concentration of doxycycline is lug/ml.
`
`Accordingly, a serum concentration of doxycycline in the range of about- 0.1 to about 0.8
`
`ug/ml is 10-80% of the minimum antimicrobial serum concentration of doxycycline. The
`
`25
`
`more preferred range is about 0.4 to about 0.7 ug/ml.
`
`To achieve this serum concentration, doxycycline can be administered by sustained
`
`release, or by intermittent administration.
`
`30
`
`In another embodiment, the tetracycline compound can be administered in an
`
`amount which results in a serum concentration between about 0.1 and 10.0 ug/ml, more
`
`14
`
`14
`
`

`

` preferably between 0.3 and 5.0 rig/ml. For example, doxycycline is administered in an
`
`amount which results in a serum concentration between about 0.1 and 0.8 ug/ml, more
`
`preferably between 0.4 and 0.7 ug/ml.
`
`Non—antimicrobial tetracycline compounds can be used in higher amounts than
`
`antimicrobial tetracyclines, while avoiding the indiscriminate killing of microbes, and the
`
`emergence of resistant microbes. For example, 6—demethyl-6-deoxy-
`
`4-de(dimethylamino)tetracycline (GMT-3) may be administered in doses of about 40 to
`
`about 200 mg/day, or in amounts that result in serum levels of about 1.5 Sug/ml to about 10
`
`10
`
`ul/ml.
`
`a?
`£33
`1’5.3
`F “3
`it.
`
`a:
`{as
`{i3
`
`a;
`up:
`
`ii:
`iii
`£23
`jut.
`
`The actual preferred amounts of tetracycline compound in a specified case will
`
`vary according to the particular compositions formulated, the mode of application, and the
`
`particular sites and subject being treated.
`
`15
`
`The tetracycline compounds can be in the form of pharmaceutically acceptable
`
`salts of the compounds. The term "pharmaceutically acceptable salt" refers to a salt
`
`prepared from tetracyCline compounds and pharmaceutically acceptable non-toxic acids or
`
`bases. The acids may be inorganic or organic acids of tetracycline compounds. Examples
`
`20
`
`of inorganic acids include hydrochloric, hydrobromic, hydroiodic, sulfuric, and phosphoric
`
`acids. Examples of organic acids include carboxylic and sulfonic acids. The radical of the
`
`organic acids may be aliphatic or aromatic. Some examples of organic acids include
`
`formic, acetic, phenylacetic, propionic, succinic, glycolic, glucuronic, maleic, furoic,
`
`glutamic, benzoic, anthranilic, salicylic, phenylacetic, mandelic, embonic (pamoic),
`
`25
`
`methanesulfonic, ethanesulfonic, panthenoic, benzenesulfonic, stearic, sulfanilic, alginic,
`
`as tartaric, citric, gluconic, gulonic, arylsulfonic, and galacturonic acids. Appropriate
`
`organic bases may be selected, for example, from N,N-dibenzylethylenediamine,
`
`chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (N-
`
`methylglucamine), and procaine.
`
`30
`
`15
`
`
`
`15
`
`

`

`Preferably, the tetracycline compounds have low phototoxicity, or is administered
`
`in an amount that results in a serum level at which the phototoxicity is acceptable.
`
`Phototoxicity is a chemically-induced photosensitivity. Such photosensitivity renders skin
`
`susceptible to damage, e. g. sunburn, blisters, accelerated aging, erythemas and eczematoid
`
`5
`
`lesions, upon exposure to light, in particular ultraviolet light. The preferred amount of the
`
`tetracycline compound produces no more phototoxicity than is produced by the
`
`administration of a 40mg total daily dose of doxycycline.
`
`Phototoxicity can be evaluated in terms of a photoirritancy factor (PIF), as
`
`10
`
`described in the examples. A PIF value of about 1.0 indicates that a compound is
`
`considered to have no measurable phototoxicity.
`
`The low phototoxic derivatives preferably have PIF values no greater than about 5,
`
`preferably no greater than about 2, more preferably no greater than about 1.5, most
`
`15
`
`preferably no greater than about 1.2, and optimally about 1.
`
`Some antimicrobial tetracyclines having low phototoxicity include, for example,
`
`minocycline and tetracyline.
`
`20
`
`Some non-antimicrobial tetracyclines having low phototoxicity include, but are not
`
`limited to, tetracycline compounds having the general formulae:
`
`STRUCTURE K
`
`25
`
`wherein: R7, R8, and R9 taken together in each case, have the following meanings:
`
`R7
`
`R8
`
`R9
`
`30
`
`and
`
`hydrogen
`hydrogen
`hydrogen
`
`amino
`palmitamide
`dimethylamino
`
`hydrogen
`hydrogen
`hydrogen
`
`l O
`
`,._.,.._.___,..__.__..H.._.,,
`
`,,
`
`,
`
`, ,WmTFW ,
`
`,,
`
`16
`
`

`

`STRUCTURE L
`
`STRUCTURE N
`
`STRUCTURE M '
`
`STRUCTURE O
`
`5
`
`10 '
`
`1 5
`
`wherein: R7, R8, and R9 taken together in each case, have the following meanings:
`
`R7
`
`hydrogen
`hydrogen
`hydrogen
`hydrogen
`
`and
`
`R8
`
`R9
`
`‘
`
`hydrogen
`hydrogen
`hydrogen
`hydrogen
`
`acetamido
`dimethylaminoacetamido
`. nitro
`amino .
`
`STRUCTURE P
`
`wherein: R7, R8, and R9 taken together are, respectively, hydrogen, hydrogen and nitro.
`
`20
`
`The tetracycline compounds are administered without administering a
`
`bisphosphonate compound. Bisphosphonates compounds are related to inorganic
`
`pyrophosphonic acid. The bisphosphonates include, as non-limiting examples,
`
`alendronate ((4-amino-l- hydroxybutylidene) bisphosphonic acid), clodronate
`
`(dichloromethane diphosphonic acid), etidronate ((l—hydroxyethylidene) diphosphanic
`
`25
`
`acid) and pamidronate ((3 —amino-1- hydroxypropylidene) bisphosphonic acid); also
`risedronate ([-hydroxy-2-(3 -pyridiny1)ethylidene] bisphosphonic acid), tiludronate, i.e.,
`
`tiludronic acid ([(4-chlorophenyl) thio] methylene] bisphosphonic acid) and zolendronate.
`
`The tetracycline compounds are administered systemically. For the purposes of
`
`30
`
`this specification, “systemic administration” means administration to a human by a
`
`method that causes the compounds to be absorbed into the bloodstream.
`
`ll
`
`17
`
`

`

`{F
`
`“a"
`
`"3?a“:““3man.1.zw,.“I'd”Siamiimir,my
`
`
`‘53”“12“”‘€5.31“51'
`s:t:m1-1?=a“...:1ii1.55my:sn!
`
`‘L
`if;an
`a
`if.
`3.3
`
`For example, the tetracyclines compounds can be administered orally by any
`
`method known in the art. For example, oral administration can be by tablets, capsules,
`
`pills, troches, elixirs, suspensions, syrups, wafers, chewing gum and the like.
`
`5
`
`Additionally, the tetracycline compounds can be administered enterally or
`
`parenterally, e.g., intravenously, intramuscularly, or subcutaneously, as injectable
`
`solutions or suspensions. Administration can also be intranasally, in the form of, for
`
`example, an intranasal spray; or transdermally, in the form of, for example, a patch.
`
`10
`
`For the pharmaceutical purposes described above, the tetracycline compounds of
`
`the invention can be formulated per se in pharmaceutical preparations optionally with a
`
`suitable pharmaceutical carrier (vehicle) or excipient as understood by practitioners in the
`
`art. These preparations can be made according to conventional chemical methods.
`
`15
`
`Examples of carriers and excipients include starch, milk, sugar, certain types of
`
`clay, gelatin, stearic acid or salts thereof, magnesium or calcium stearate, talc, vegetable
`
`fats or oils, gums and glycols.
`
`Inthe embodiment in which the tetracycline compound ‘is a non-antimicrobial
`
`20
`
`tetracycline compound, administration can include topical application. Particular non-
`
`antimicrobialtetracycline compounds have only limited biodistribution, e. g. CMT-S. In
`
`such cases, topical application is the preferred method of administration of the compound.
`
`Carrier compositions deemed to be suited for topical use include gels, salves,
`
`25
`
`lotions, creams, ointments and the like. The non—antimicrobial tetracycline compound can
`
`also be incorporated with a support base or matrix or the like which can be directly applied
`
`to skin.
`
`Topical application of non-antimicrobial tetracycline compounds in amounts of up
`
`30
`
`to about 25% (w/w) in a vehicle are effective in treating acne while not inducing
`
`1,2
`
`18
`
`18
`
`

`

`significant toxicity in the human. Amounts of from about 0.1% to about 10% are
`
`preferred.
`
`Combined or coordinated topical and systemic administration of the tetracycline
`
`5
`
`compounds is also contemplated under the invention. For example, a non-absorbable non-
`
`antimicrobial tetracycline compound can be administered topically, while a tetracycline
`
`compound capable of substantial absorption and effective systemic distribution in a human
`
`can be administered systemically.
`
`10
`
`The tetracycline compounds are prepared by methods known in the art. For
`
`:1;
`
`ll;um!3
`
`fl:
`
`:1.
`.
`iii}
`E‘aa
`up?
`
`$33W
`‘iu:
`
`£33
`13?
`{:33
`
`15
`
`20
`
`25
`
`30
`
`example, natural tetracyclines may be modified without losing their antibiotic properties,
`
`although certain elements of the structure must be retained. The modifications that may
`
`and may not be made to the basic tetracycline structure have been reviewed by Mitscher in
`
`The Chemistry of Tetracyclines, Chapter 6, Marcel Dekker, Publishers, New York (1978).
`According to Mitscher, the substituents at positions 5—9 of the tetracycline ring system
`may be modified without the complete loss of antibiotic properties. Changes to the basic
`ring system or replacement of the substituents at positions 1-4 and 10-12, however,
`
`generally lead to synthetic tetracyclines with substantially less or effectively no
`
`antimicrobial activity.
`
`Further methods of preparing the tetracycline compounds are described in the
`
`examples.
`
`EXAMPLES
`
`The following examples serve to provide fiirther appreciation of the inVention but
`
`are not meant in any way to restrict the effective scope of the invention.
`
`Preparation of Compounds
`
`EXAMPLE 1
`
`4-Dedimethylamino-7-dimethylamino—6-demethyl-6-deoxy-9-nitrotetracycline sulfate
`
`13
`
`19
`
`
`
`19
`
`

`

`
`
`
`
`To a solution of one millimole of 4-dedimethylamino-7-dimethylamino—6-
`
`demethyl-6-deoxytetracycline in 25 ml of concentrated sulfuric acid at 0°C was added
`
`1.05 mmole of potassium nitrate. The resulting solution was stirred at ice bath
`
`temperature for 15 minutes and poured in one liter of cold etherwith stirring. The
`
`precipitated solid was allowed to settle and the majority of solvent decanted. The
`
`remaining material wasfiltered through a sintered glass funnel and the collected solid was
`
`washed'well with cold ether. The product was dried in a vacuum desiccator overnight.
`
`10
`
`EXAMPLE 2
`
`9-amino-4-dedimethylamino—7—dimethylamino-6-demethyl-6-deoxytetracycline
`
`sulfate
`
`gar
`a?
`
`To a solution of 300 mg of the 9-nitro compound from example 1, in 30 m1 of
`
`15
`
`ethanol was added 50 mg of PtOz. The mixture was hydrogenated at atmospheric pressure
`until the theoretical amount of hydrogen was absorbed. The system is flushed with I
`
`nitrogen, the catalyst PtOz is filtered and the filtrate added dropwise to 300 m1 of ether.
`
`The product that separates is filtered and dried in a vacuum desiccator.
`
`20
`
`EXAMPLE 3
`
`9-Acetamido-4-dedimethylamino-7-dimethylamino-6-demethy1—6—deoxytetracycline
`
`sulfate
`
`To a well stirred cold solution of 500 mg of 9-amino-4-dedimethylamino-7-
`
`25
`
`dimethylamino-6-demethyl-6-deoxytetracycline sulfate from example 2, in 2.0 ml of 1.3-
`
`dimethyl-2-imidazolidinone, 500 mg of sodium bicarbonate was added followed by 0.21
`
`ml of acetyl chloride. The mixture is stirred at room temperature for 30 minutes, filtered
`
`and the filtrate was added dropwise to 500 m1 of ether. The product that separated was
`
`filtered and dried in a vacuum desiccator.
`
`30
`
`14
`
`20
`
`20
`
`

`

`
`
`EXAMPLE 4
`
`
`
`4-Dedimethylamino-7-dimethylamino-6-demethyl-6-deoxy-9-diazoniumtetracycline
`
`sulfate
`
`To a solution of 0.5 g of 9-amino-4-dedimethylamino-7-dimethy1amino-6-
`
`demethyl-6-deoxytetracycline sulfate, from example 2,
`
`in 10 ml of 0.1N hydrochloric acid
`
`in methanol cooled in an ice bath, 0.5 m1 of n-butyl nitrite was added. The solution was
`
`stirred at ice bath temperature for 30 minutes and then poured into 250 m1 of ether. The
`
`product that separated was filtered, washed with ether and dried in a vacuum desiccator.
`
`EXAMPLE 5
`
`9-Azido-4-dedimethylamino-7-dimethylamino-6-demethyl-6-deoxytetracycline
`
`sulfate
`
`To a solution of 0.3 mole of 4-dedimethylamino-7-dimethylamino-6-demethyl-6-
`
`deoxy-9-diazoniumtetracycline sulfate, from example 4, 10 ml of 0.1 N methanolic
`hydrogen chloride was added 0.33 mmole of sodium azide. The mixture was stirred at
`room temperature for 1.5 hours. The reaction mixture was then poured into 200 ml of
`
`ether. The product that separated was filtered and dried in a vacuum desiccator.
`
`EXAMPLE 6
`
`9-Amino—8-chloro—4-dedimethylamino-7-dimethylamino-6—demethy1-6-deoxy-
`
`tetracycline sulfate
`
`10
`
`15
`
`20
`
`25
`
`One gram of 9-azido-4-dedimethylamino—7-dimethylamino-6-demethyl-6—
`
`deoxytetracycline hydrochloride, from example 4, was dissolved in 10 m1 of concentrated
`
`sulfuric acid saturated with HCL at 0°C. The mixture was stirred at ice bath temperature
`
`for 1.5 hours and then slowly added dropwise to 500 m1 of cold ether. The product that
`
`3O
`
`separated was filtered, washed with ether and dried in a vacuum desiccator.
`
`15
`
`21
`
`
`
`21
`
`

`

`
`
`W
`
`4-Dedimethylamino-7—dimethylamino-6-demethyl-6-deoxy—9-ethoxythiocarbony1thio-
`
`tetracycline sulfate
`
`A sol