`
`1111111111111111111111010!!),1110t !11011121111111111111111111111111
`
`(12) United States Patent
`Cavaleri
`
`(10) Patent No.: US 10,945,970 B2
`Mar. 16, 2021
`(45) Date of Patent:
`
`(54)
`
`CURCUMIN-BASED COMPOSITIONS AND
`METHODS OF USE THEREOF
`
`(71)
`
`Applicant: Franco Cavaleri, Surrey (CA)
`
`(72)
`
`Inventor: Franco Cavaleri, Surrey (CA)
`
`* )
`
`Notice:
`
`Subject to any disclaimer, the term of this
`patent is extended or adjusted under 35
`U.S.C. 154(b) by 207 days.
`
`(21)
`
`Appl. No.: 16/111,502
`
`(22)
`
`Filed:
`
`Aug. 24, 2018
`
`(65)
`
`Prior Publication Data
`
`US 2018/0360774 Al
`
`Dec. 20, 2018
`
`Related U.S. Application Data
`
`(63)
`
`application
`of
`Continuation-in-part
`PCT/CA2018/050275, filed on Mar. 8, 2018.
`
`No.
`
`(60)
`
`Provisional application No. 62/469,554, filed on Mar.
`10, 2017.
`
`(51)
`
`Int. Cl.
`A61K 36/00
`A61K 31/12
`A61K 9/48
`A61P 3/06
`A61K 36/9066
`(52) U.S. Cl.
`CPC
`
`(2006.01)
`(2006.01)
`(2006.01)
`(2006.01)
`(2006.01)
`
` A61K 31/12 (2013.01); A61K 9/485
`(2013.01); A61K 9/4858 (2013.01); A61K
`
`9/4866 (2013.01); A61P 3/06 (2018.01); A61K
`36/9066 (2013.01); A61K 2300/00 (2013.01)
`(58) Field of Classification Search
`None
`See application file for complete search history.
`
`(56)
`
`References Cited
`
`U.S. PATENT DOCUMENTS
`
`2008/0193573 A1*
`
`8/2008 Gow
`
`2015/0366815 A1* 12/2015 Teles
`
`* cited by examiner
`
`Primary Examiner
`
` Qiuwen Mi
`
` A61P 25/28
`424/756
` A61K 9/485
`514/769
`
`ABSTRACT
`(57)
`Methods and compositions for increasing LDL receptor
`levels in a plurality of cells are disclosed. For example, the
`methods include providing to the cells a curcumin compo-
`sition that is at least 15% curcumin II or at least 5%
`curcumin III, in order to increase LDL receptor levels. In
`addition, compositions for increasing LDL receptor levels in
`a plurality of cells are disclosed, which include a curcumin
`composition consisting of at least 15% curcumin II or at
`least 5% curcumin III (or combinations of such curcumi-
`noids), along with a pharmaceutically-acceptable solvent,
`filler, or carrier. Still further, methods and compositions for
`decreasing MSK1 levels in a plurality of cells are disclosed,
`which involve administering a curcumin III-enriched com-
`position to the cells.
`
`7 Claims, 12 Drawing Sheets
`
`SAB1001
`U.S. Pat. No. 10,945,970
`
`

`

`U.S. Patent
`
`Mar. 16, 2021
`
`Sheet 1 of 12
`
`US 10,945,970 B2
`
`Empower
`
`Curcumtn Extract
`
`—
`
`Curcumin
`
`—
`
`— Curcum n
`
`1
`
`Curcurnin
`
`•LE
`
`50
`
`0
`0
`
`74%
`
`,Isssame
`
`0
`
`0
`
`0
`
`0
`00
`
`Concentration
`
`FIGURE 1
`
`

`

`U.S. Patent
`
`Mar. 16, 2021
`
`Sheet 2 of 12
`
`US 10,945,970 B2
`
`MU Assay
`BV2 (rnicrogtiai} Cell Line
`
`moircir, Mpower
`Curcumin Extract
`Curcumin l
`
`j
`
`
`
`Curtumin tit
`
`120
`
`100
`
`80
`
`60
`
`40
`
`20
`
`S
`
`1.0
`Concentration in ughni
`
`20
`
`40
`
`80
`
`FIGURE 2
`
`

`

`U.S. Patent
`
`Mar. 16, 2021
`
`Sheet 3 of 12
`
`US 10,945,970 B2
`
`1.4
`43 • 1.2
`
`u 1
`0.8
`2 0.6
`C▪ • 0 4
`P5▪ 0.2
`a.
`0
`
`0 65 Mal protein
`iu3s
`glia 3 iter) .... ............................
`............................................. ... t.. . .............. .........
`in
`8V2
`ement IVISO
`
`trot
`
`111 NI
`111111 1111
`11111 1111 1111
`III III III IIIII
`
`111 WIN
`Ill 11111 III III
`111111 111 Nil
`1111
`
`sv
`.4‘
`
`0
`
`ft) Q`)
`
`\
`
`4 )
`
`
`Q
`
`et`v
`‘-'
`
`
`
`it)
`„,...N.,
`444kY
`
`qc)
`48,se
`
`
`4.1 %,..
`0
`\
`
`,\
`4t )
`,,
`.'7
`
`4 1
`\,
`',.41
`
`(A
`• 44.
`
`45>
`
`4%
`(it)
`
`FIGURE 3
`
`

`

`U.S. Patent
`
`Mar. 16, 2021
`
`Sheet 4 of 12
`
`US 10,945,970 B2
`
`Nuc-p65/Total Protein
`(1.00mcgirni IPS In BV2 Microglia - 3 'ter)
`Percent D 50 cor troi
`
`6
`
`5
`
`ie 1
`
`41
`‘.
`
`Ati)
`V
`
`4 7
`CI\*
`
`0
`%re
`• "''6.1`
`szt+
`
`\*$‘
`
`0
`...%,
`
`\..Q
`C"
`4
`\
`
`%1 7
`
`')
`4
`'V
`4
`
`(>
`
`'61/4
`
`0 #1
``2.
`
`sk
`0 )
`
`4*6Z
`
`CAP
`
`FIGURE 4
`
`

`

`U.S. Patent
`
`Mar. 16, 2021
`
`Sheet 5 of 12
`
`US 10,945,970 B2
`
`Cyto-p65 Ser276/Total Protein
`
`(1:00mcgimi IPS in 8V2 Microglia - alter)
`Percent DIMS° control
`
`1.4
`
`I
`
`0.8
`
`0.6
`
`0.4
`
`0.2
`
`0
`
`Percent IMMO Control
`
`C
`'2.
`
`Cf\Y
`
`\ C .>
`V
`
`4,f)
`Q.,
`
`(.2\‘'
`
`FIGURE 5
`
`

`

`U.S. Patent
`
`Mar. 16, 2021
`
`Sheet 6 of 12
`
`US 10,945,970 B2
`
`1.4
`
`1.2
`
`g
`
`1
`
`U
`0 0.8
`
`0.2
`
`0
`
`Nuc-p65 Ser276/Total Protein
`
`(100mcgiml IPS in 8W Microglia - 3 Iter)
`
`Percent DMSO control
`
`111
`
`Aes,'
`
`cq4)
`
`o
`4')
`
`' '':..k
`
`e
`
`.
`
`S
`Sik .
`
`ke44
`
`C
`
`Or
`'iC'
`
`cec\
`
`FIGURE 6
`
`

`

`U.S. Patent
`
`Mar. 16, 2021
`
`Sheet 7 of 12
`
`US 10,945,970 B2
`
`1.4
`
`1.2
`
`
`
`T
`
`Cyto-MSKI Total/Total Protein
`(100mceml LPS in BV2 Microglia - 3 ter
`Percent DIVISO Control -
`
`O
`
`* S>
`
`
`
`q 4"
`
`.14 )
`cr
`
`Ni l
`o
`
`47q
`
`,s4..N
`
`,A4‘ .
`'ter
`
`s k ' •
`
`' ' : .` >'
`
` '
`
`CP
`X•.>
`
`45'd
`
`,,\* et`
`
`0
`
`FIGURE 7
`
`

`

`U.S. Patent
`
`Mar. 16, 2021
`
`Sheet 8 of 12
`
`US 10,945,970 B2
`
`Nuc MSKI Totaltrotal Protein
`(100mcgimi IPS in BV2 Microglia - 3 Iter)
`Percent DIVISO Control
`
`2
`
`8
`0
`
`g 03
`
`,...p
`
`\-\
`
`.f.,
`
`4-)
`
`FIGURE 8
`
`

`

`U.S. Patent
`
`Mar. 16, 2021
`
`Sheet 9 of 12
`
`US 10,945,970 B2
`
`1.2
`
`0
`
`MSKI Ser 376/1otal Protein
`tit .4 eml LPS in..B 2 Micri, lia -3 tier
`Percent
`.s co
`
`\*
`
`0§‘ 0§
`
`41\
`
`N..
`
`0
`
`s.'
`
`co
`
`FIGURE 9
`
`

`

`U.S. Patent
`
`Mar. 16, 2021
`
`Sheet 10 of 12
`
`US 10,945,970 B2
`
`Nuo.IVISK1Ser 316/Tota I Protein
`
` tioorneginaps in 6V2 Microglia -3 itOr
`
`
`Percent •DA/1S0 control
`
`1.6
`
`1.4
`
`43c 1.2
`c..)
`0
`11 0.8
`
`,
`
`,
`
`T
`
`ger
`0 ,
`
`4 t, k.CY
`,
`<4,4.
`
`i k'''
`Xlk
`,0 *
`
`to-
`(.,
`
`„,•`'
`(A
`\(,.,
`.4>
`4,cio
`
`FIGURE 10
`
`

`

`U.S. Patent
`
`Mar. 16, 2021
`
`Sheet 11 of 12
`
`US 10,945,970 B2
`
`LDL receptor expression by HepG2 cells after treating
`with curcuminoids (pooled, n=5)
`
`[CELLRANGEI
`
`ICELLRANGE)
`
`[cELLRANGEI
`
`T
`!,........t.A
`,z.........,...,
`NzZzZ ". .....
`
`ZZZ‘Z‘Z Z Z.•
` ZZZZZZZ Z Z.•
`ZZZZZZZ Z Z.•
`ZZZZZZZ Z Z.•
`ZZZZZ‘ Z % %
`
`E.)
`
`....... <
`.
`
`-.1
`
`0.6
`
`0.4
`
`0.2
`
`[CELLRANGEJ
`
`........Z
`
`
`
`M1 M1 M1 ......
`
`,
`,
`,
`,
`,
`'
`
`M1...M1
`
`....M1
`....M1
`
`....M1 ....
`....M1 ....
`.. ....
`....M1 ..
`.....M1 ....
`....M1 ..
`....M1 ....
`....M1 ....
`....M1 ....
`....M1 ....
` ....M1 ....
`....M1 ....
`.. .. Z
`.. .. Z
`\Z‘ M1 M1
`Z........%
`
`....M1
`....M1
`....M1
`....M1
`
`Curcurnin i
`
`Curcurnin t1
`
`Curcumin 111
`
`DMSO
`
`Control
`
`FIGURE 11
`
`

`

`U.S. Patent
`
`Mar. 16, 2021
`
`Sheet 12 of 12
`
`US 10,945,970 B2
`
`LD receptor expression by ilepS2 cells after
`treating with curcuminoids (3Ougi 1 n
`
`ce OS
`0.6
`0.4
`
`Curcurain
`
`Curcuri : 1>
`
`:rcun#€
`
`11
`
`0MSO
`
`Contmi
`
`FIGURE 12
`
`•
`

`

`US 10,945,970 B2
`
`1
`CURCUMIN-BASED COMPOSITIONS AND
`METHODS OF USE THEREOF
`
`CROSS-REFERENCE TO RELATED
`APPLICATIONS
`
`This application is a continuation-in-part application of
`PCT application serial number PCT/CA2018/050275, filed
`on Mar. 8, 2018, which claims priority to, and incorporates
`by reference, U.S. provisional patent application 62/469,
`554, filed on Mar. 10, 2017.
`
`FIELD OF THE INVENTION
`
`The field of the present invention relates to certain cur-
`cumin-containing compositions and methods of use thereof,
`which can be used to increase low-density lipoprotein cho-
`lesterol (LDL) receptor expression levels and thereby lower
`LDL levels in a plurality of cells or subject. In addition, the
`field of the present invention relates to certain curcumin-
`containing compositions and methods of use thereof, which
`can be used to modulate MSK1 production and thereby
`ameliorate a variety of health conditions.
`
`BACKGROUND OF THE INVENTION
`
`The health benefits of curcumin, particularly whole tur-
`meric extract, are known and have been demonstrated by
`researchers in recent years. However, several challenges
`continue to exist, with respect to the formulation of cur-
`cumin-based pharmaceuticals and dietary supplements.
`More specifically, the most common source of curcumin, the
`Indian spice turmeric (a member of Zingiberaceae), does not
`contain a sufficient amount of curcumin to provide an
`efficacious dose to a subject. In fact, the therapeutic benefits
`provided by natural curcumin extracts have been relatively
`modest, very inconsistent, and not well understood. Accord-
`ingly, there is a continuing need for improved curcumin-
`based formulations, which address these current challenges.
`The present invention, as described further below,
`addresses many of the foregoing challenges.
`
`SUMMARY OF THE INVENTION
`
`0
`
`2
`plurality of cells which results in lower LDL levels in the
`cells and/or subject (which produces a number of therapeutic
`and health benefits).
`According to further aspects of the present invention,
`5 LDL receptor-modulating therapeutic compositions are dis-
`closed that comprise a curcumin composition that includes
`at least 15% (w/v) curcumin II, along with a pharmaceuti-
`cally acceptable solvent, filler, or carrier. The invention
`provides that while the curcumin composition employed
`may comprise 15% (w/v) curcumin II, in certain preferred
`embodiments, the curcumin composition employed may
`comprise at least 30% (w/v) curcumin II. Still more prefer-
`ably, the invention provides that the curcumin composition
`15 may comprise at least 50% (w/v) curcumin II, at least 70%
`(w/v) curcumin II or, even more preferably, at least 90%
`(w/v) curcumin II.
`According to still further aspects of the present invention,
`LDL receptor-modulating therapeutic compositions are dis-
`20 closed that include a curcumin composition that includes at
`least 5% (w/v) curcumin III and a pharmaceutically accept-
`able solvent, filler, or carrier. The invention provides that
`while the curcumin composition employed may comprise
`5% (w/v) curcumin III, in certain preferred embodiments,
`25 the curcumin composition employed may comprise at least
`30% (w/v) curcumin III. Still more preferably, the invention
`provides that the curcumin composition may comprise at
`least 50% (w/v) curcumin III, at least 70% (w/v) curcumin
`III or, even more preferably, at least 90% (w/v) curcumin III.
`30 According to additional aspects of the invention, LDL
`receptor-modulating therapeutic compositions are disclosed
`that include a combination of the curcumin II and curcumin
`III enriched compositions described above.
`According to additional aspects of the present invention,
`35 certain curcumin III enriched compositions described herein
`may be used to further modulate mitogen- and stress-
`activated protein kinase 1 (MSK1), which is a nuclear kinase
`that plays a significant role in transcription regulation
` produces a number of therapeutic and health ben-
`(whichs).
`40 efit
`The above-mentioned and additional features of the pres-
`ent invention are further illustrated in the Detailed Descrip-
`tion contained herein.
`
`BRIEF DESCRIPTION OF THE FIGURES
`
`According to certain aspects of the present invention, 45
`methods for increasing low-density lipoprotein cholesterol
`(LDL) receptor expression levels in a plurality of cells are
`disclosed (and, by extension, methods of reducing LDL
`levels in the cells). In certain embodiments, the methods
`comprise providing to the cells a composition that includes
`an effective and enriched amounts of curcumin II, curcumin
`III, or a combination of curcumin II and curcumin III. In
`certain embodiments, the composition is at least 15% (w/v)
`curcumin II or, preferably, at least 30% (w/v) curcumin II or,
`more preferably, at least 50% (w/v) curcumin II or, even
`more preferably, at least 70% (w/v) curcumin II, such as at
`least 90% (w/v) curcumin II. In other embodiments, the
`composition is at least 5% (w/v) curcumin III or, preferably,
`at least 30% (w/v) curcumin III or, more preferably, at least
`50% (w/v) curcumin III or, even more preferably, at least
`70% (w/v) curcumin III, such as at least 90% (w/v) cur-
`cumin III. In still further embodiments, the composition
`includes a combination of the curcumin II and curcumin III
`enriched compositions summarized above. As described and
`exemplified further herein, the invention provides that
`administration of such curcumin II and curcumin III
`enriched compositions elevates LDL receptor levels in a
`
`55
`
`FIG. 1: MTT assay results demonstrating HEK293 cell
`survival of approximately 80% for all three curcuminoids
`(ranging from 20 to 22 1,tg/mL of the applicable curcumi-
`so noid).
`FIG. 2: MTT assay results demonstrating BV2 cell sur-
`vival of approximately 80% for all three curcuminoids
`(ranging from 20 to 22 1,tg/mL of the applicable curcumi-
`noid).
`FIG. 3: measurements of cytoplasmic NFkB-p65 protein
`levels relative to total protein concentration in BV2 cell lines
`provided with curcumin extract, curcuminoid I, curcuminoid
`II, curcuminoid III, and controls.
`FIG. 4: measurements of nuclear NFkB-p65 protein levels
`60 relative to total protein concentration in BV2 cell lines
`provided with curcumin extract, curcuminoid I, curcuminoid
`II, curcuminoid III, and controls.
`FIG. 5: measurement of cytoplasmic NFkB p65 that is
`phosphorylated at the serine 276 phosphosite, relative to
`65 total protein concentration in BV2 cell lines provided with
`curcumin extract, curcuminoid I, curcuminoid II, curcumi-
`noid III, and controls.
`
`

`

`US 10,945,970 B2
`
`3
`FIG. 6: measurement of nuclear NFkB p65 that is phos-
`phorylated at the serine 276 phosphosite, relative to total
`protein concentration in BV2 cell lines provided with cur-
`cumin extract, curcuminoid I, curcuminoid II, curcuminoid
`III, and controls.
`FIG. 7: measurement of cytoplasmic MSK1 protein levels
`relative to total protein concentration in BV2 cell lines
`provided with curcumin extract, curcuminoid I, curcuminoid
`II, curcuminoid III, and controls.
`FIG. 8: measurement of nuclear MSK1 protein levels
`relative to total protein concentration in BV2 cell lines
`provided with curcumin extract, curcuminoid I, curcuminoid
`II, curcuminoid III, and controls.
`FIG. 9: measurement of cytoplasmic MSK1 that is phos-
`phorylated at the serine 376 phosphosite, relative to total
`protein concentration in BV2 cell lines provided with cur-
`cumin extract, curcuminoid I, curcuminoid II, curcuminoid
`III, and controls.
`FIG. 10: measurement of nuclear MSK1 that is phospho-
`rylated at the serine 376 phosphosite, relative to total protein
`concentration in BV2 cell lines provided with curcumin
`extract, curcuminoid I, curcuminoid II, curcuminoid III, and
`controls.
`FIG. 11: measurement of LDL receptor expression levels
`in HepG2 cells after treatment with 22 ng/mL curcuminoid
`I, curcuminoid II, curcuminoid III, and controls.
`FIG. 12: measurement of LDL receptor expression levels
`in HepG2 cells after treatment with 30 ng/mL curcuminoid
`I, curcuminoid II, curcuminoid III, and controls.
`
`DETAILED DESCRIPTION OF THE
`INVENTION
`
`10
`
`4
`invention provides that curcumin III (and not curcuminoids
`I and II) can be used to selectively and efficaciously inhibit
`cytoplasmic and nuclear MSK1 production, the inhibition of
`MSK1 serine376 phosphorylation, and inhibition of the
`5 recruitment of MSK1 at inflammatory gene promotors. The
`curcumin III compositions described herein and related
`methods of using such compositions provide a major step
`in the transactivation regulation of downstream transcription
`factors that are key to cell survival and recruitment of
`inflammatory and immune system events. For example, as
`demonstrated in the Examples below, the ability of the
`curcumin III compositions described herein to inhibit MSK1
`production (or otherwise significantly reduce MSK1 levels)
`15 indicates that such compositions may also (indirectly) be
`used to modulate NFkB (nuclear factor kappa-light-chain-
`enhancer of activated B cells)
`the aberrant expression and
`transactivation of which has been linked to cancer, inflam-
`mation, and autoimmune diseases. The curcumin III com-
`20 positions (and related methods) of the present invention
`provide improved efficacy, reliability, and drug target selec-
`tivity, relative to natural curcum in extracts.
`A natural curcumin extract comprises a mixture of cur-
`cumin I, desmethoxycurcumin (curcumin II), and bisde-
`25 methoxycurcumin (curcumin III). The term curcumin refers
`to the principal curcuminoid in the Indian spice turmeric
`plant (a member of Zingiberaceae). The IUPAC name for the
`curcumin
`I molecule
`is (1E,6E)-1,7-Bis(4-hydroxy-3-
`methoxypheny1)-1,6-heptadiene-3,5-dione. Although cur-
`30 cumin I may exist in several different tautomeric forms, the
`enol form is illustrated below:
`
`The following will describe, in detail, several preferred
`embodiments of the present invention. These embodiments 35
`are provided by way of explanation only, and thus, should
`not unduly restrict the scope of the invention. In fact, those
`of ordinary skill in the art will appreciate upon reading the
`present specification and viewing the present drawings that
`the invention teaches many variations and modifications, 40
`and that numerous variations of the invention may be
`employed, used and made without departing from the scope
`and spirit of the invention.
`According to certain preferred embodiments, the present
`invention includes certain curcumin-enriched compositions 45
`(and methods of using such compositions). More particu-
`larly, the present invention includes certain compositions
`that contain elevated and concentrated levels of (1) cur-
`cumin II (relative to the amount of curcumin II found in
`natural curcumin extract); (2) curcumin III (relative to the 50
`amount of curcumin III found in natural curcumin extract);
`or (3) a combination of curcumin II and curcumin III
`(relative to the amounts of curcumin II and curcumin III
`found in natural curcumin extract). Notably, the LDL recep-
`tor-modulating compositions described herein will prefer-
`ably exclude curcumin I. The invention provides that such
`compositions can be used to increase low-density lipopro-
`tein cholesterol (LDL) receptor expression levels in a plu-
`rality of cells (which, in turn, results in lower LDL levels in
`a subject and ameliorates a variety of associated health 60
`conditions and/or impart one or more associated health
`benefits).
`In addition, the invention provides that certain curcumin
`III enriched compositions described herein may be used to
`modulate mitogen- and stress-activated protein kinase 1
`(MSK1), which is a nuclear kinase that plays a significant
`role in transcription regulation. As described below, the
`
`55
`
`65
`
`HO
`
`S
`
`OH
`
`CH3
`
`0
`
`OH
`
`CH3
`
`The IUPAC name for the desmethoxycurcumin (curcumin
`II) molecule is (1E,6E)-1-(4-Hydroxy-3-methoxypheny1)-7-
`(4-hydroxyphenyl)hepta-1,6-diene-3,5-dione, and has the
`chemical structure shown below:
`
`0 0
`
`The IUPAC name for bis-desmethoxycurcumin (cur-
`cumin III) that is used in the compositions and methods of
`the present invention is (1E,6E)-1,7-bis(4-hydroxyphenyl)
`hepta-1,6-diene-3,5-dione, and has the chemical structure
`shown below:
`
`0
`
`

`

`US 10,945,970 B2
`
`5
`According to certain preferred embodiments, the inven-
`tion provides that curcumin II and curcumin III may be
`extracted from turmeric plant rhizome (Curcuma longa) and
`subsequently concentrated to the desired levels. Alterna-
`tively, the invention provides that the curcumin II and
`curcumin III molecules may be chemically synthesized and
`used to formulate a therapeutic composition described
`herein. As explained below, the desired concentration of
`curcumin II is at least 15%, 30%, 50%, 70%, or 90% (w/v)
`curcumin II, while the desired concentration of curcumin III
`is at least 5%, 30%, 50%, 70%, or 90% (w/v) curcumin III.
`According to certain preferred embodiments of the pres-
`ent invention, methods for increasing LDL receptor expres-
`sion levels (and thereby lowering LDL) in a plurality of cells
`(and subject) are provided. In such embodiments, the meth-
`ods include providing to the cells (or administering to a
`biological system that comprises a plurality of cells) an
`effective amount of a LDL receptor-modulating curcumin
`composition that is (1) at least 15% curcumin II; (2) at least
`5% curcumin III; or (3) a combination of (1) and (2).
`According to additional preferred embodiments of the pres-
`ent invention, methods for inhibiting MSK1 serine376 phos-
`phorylation in a plurality of cells are provided. Such meth-
`ods include providing to the cells (or administering to a
`biological system that comprises a plurality of cells) an
`effective amount of a curcumin composition that is at least
`5% curcumin III.
`The "effective amount" of a LDL receptor-modulating
`curcumin composition will preferably be sufficient to sig-
`nificantly increase LDL receptor expression levels (such as
`by at least 10% relative to a control cell line or, even more
`preferably, by at least 20% relative to a control cell line), to
`thereby reduce the amount of LDL in the target cells (and
`subject). Similarly, the "effective amount" of a MSK1-
`modulating curcumin composition will preferably be suffi-
`cient to significantly reduce the amount of MSK1 protein
`being expressed in the target cells (such as by at least 10%
`relative to a control cell line or, even more preferably, by at
`least 20% relative to a control cell line).
`According to certain preferred embodiments of the pres-
`ent invention, methods for preventing and/or ameliorating
`the effects of certain diseases associated with high choles-
`terol (LDL) levels are provided. Such methods generally
`include providing to a subject an effective amount of the
`curcumin II and/or curcumin III enriched compositions
`described herein. Non-limiting examples of such diseases
`include cardiovascular diseases (including heart disease,
`stroke, peripheral vascular disease, atherosclerosis, arterio-
`sclerosis, and serum LDL elevation), diabetes, and high
`blood pressure.
`According to yet further preferred embodiments of the
`present invention, methods for preventing and/or ameliorat-
`ing the effects of an adverse medical condition in which
`MSK1 is implicated are provided, including glucocorticoid-
`resistant inflammatory diseases and chemotherapy-resistant
`cancers. In such embodiments, the methods include provid-
`ing to a subject a curcumin composition that is at least 5%
`curcumin III (or, alternatively, at least 30%, 50%, 70%, or
`90% (w/v) curcumin III). According to certain related
`embodiments of the present invention, therapeutic compo-
`sitions are provided that include a curcumin composition
`consisting of at least 5% (w/v) curcumin III; optionally,
`glucocorticoids; and a pharmaceutically acceptable solvent,
`filler, or carrier. As used herein, "glucocorticoids" refers to
`certain steroid hormones that are known to bind to gluco-
`corticoid receptors (RCEs). Non-limiting examples of glu-
`cocorticoids include: cortisol, cortisone, prednisone, pred-
`
`6
`dexamethasone,
`methylprednisolone,
`nisolone,
`betamethasone, triamcinolone, beclometasone, fludrocorti-
`sone, deoxycorticosterone, and aldosterone. In the foregoing
`embodiments of the invention, while the curcumin compo-
`5 sition employed may comprise 5% (w/v) curcumin III, in
`certain preferred embodiments, the curcumin composition
`employed may comprise at least 30% (w/v) curcumin III.
`Still more preferably, the invention provides that the cur-
`cumin composition may comprise at least 50% (w/v) cur-
`io cumin III, at least 70% (w/v) curcumin III or, even more
`preferably, at least 90% (w/v) curcumin III—depending on
`the desired potency.
`The invention provides that the concentrated forms of the
`curcumin III based compositions (and methods of using such
`15 compositions) described herein exhibit many benefits
`for
`humans, canines, cats and equine. First, as demonstrated
`below and described herein, the invention provides that
`elevated levels of curcumin III will selectively inhibit MSK1
`production, which thereby produces desirable anti-inflam-
`2o matory activity. In addition, the invention provides that the
`compositions and methods described herein may be used for
`therapeutic nutrition; anti-inflammatory therapy for autoim-
`mune disease and other chronic and acute inflammatory
`ailments; treatment of pain, swelling and inflammation;
`25 nutritional supplementation; superbug treatments; and anti-
`microbial, antifungal, antibacterial, and antiviral therapies.
`Still further, according to certain additional embodiments,
`the present invention encompasses therapeutic compositions
`(and methods of use thereof) that include a curcumin com-
`30 position consisting of at least 15% curcumin II, along with
`a pharmaceutically acceptable solvent, filler, or carrier. In
`such embodiments, as described above, the curcumin II-
`enriched compositions can be used to increase LDL receptor
`expression levels and thereby lower LDL levels in target
`35 cells and/or a subject. Indeed, as shown in the Example
`below, curcumin II-enriched and curcumin III-enriched
`compositions can be used to increase LDL receptor expres-
`sion levels (and, therefore, lower LDL levels in target cells
`and/or a subject).
`In certain specific embodiments, the compositions and
`methods described herein may also be used to ameliorate the
`effects of autoimmune diseases (and other inflammatory
`conditions), such as rheumatoid arthritis, colitis, non-spe-
`cific inflammatory bowel diseases, crohn's disease, lupus,
`45 multiple sclerosis, psoriasis, type-I diabetes, diabetes, myo-
`carditis, thyroiditis, uveitis, systemic lupus erythromatosis,
`myasthenia and gravis. Furthermore, the compositions and
`methods described herein may be used to ameliorate the
`effects of autoimmune syndromes, such as the sources of
`so immune-mediated
`inflammation
`(which can promote
`chronic inflammation, Alzheimer's, asthma, allergies, obe-
`sity, chronic fatigue, fibromyelia, premature aging, and
`general memory impediments). Still further, the composi-
`tions and methods may be used for the purpose of perfor-
`55 mance enhancement; recovery from physical exercise; and
`to help neutralize lactic acid, oxidation and associated
`inflammatory responses to workload to improve recovery
`rate, anabolism, reduce post-workout soreness and associ-
`ated fatigue (and allow for repeat workout sessions earlier
`60 than could otherwise be executed in typical workout and
`training cycles).
`The invention provides that the compositions described
`herein may be administered in any desired and effective
`manner, e.g., as pharmaceutical compositions or nutritional
`65 supplements for oral ingestion. More particularly, for
`example, pharmaceutically acceptable compositions or
`nutritional supplements of the invention may comprise one
`
`40
`
`

`

`US 10,945,970 B2
`
`Each acceptable carrier used in a pharmaceutical compo- 2 o
`
`8
`7
`particular, cottonseed, groundnut, corn, germ, olive, castor
`or more of the compositions described herein with one or
`and sesame oils), glycerol, tetrahydrofuryl alcohol, polyeth-
`more acceptable carriers. Regardless of the route of admin-
`ylene glycols and fatty acid esters of sorbitan; (23) propel-
`istration selected, the compositions may be formulated into
`lants, such as chlorofluorohydrocarbons and volatile unsub-
`acceptable dosage forms by conventional methods known to
`stituted hydrocarbons, such as butane and propane; (24)
`those of skill in the art. For example, acceptable carriers 5
`antioxidants; (25) agents which render the formulation iso-
`include, but are not limited to, sugars (e.g., lactose, sucrose,
`tonic with the blood of the intended recipient, such as sugars
`mannitol, and sorbitol), silicon dioxide, starches, cellulose
`and sodium chloride; (26) thickening agents; (27) coating
`preparations (such as microcrystalline cellulose), calcium
`materials, such as lecithin; (28) vitamins and minerals; (29)
`phosphates (e.g., dicalcium phosphate, tricalcium phosphate
`proteins that carry therapeutic or nutritional benefits, such as
`and calcium hydrogen phosphate), sodium citrate, water, 10
`whey protein and other milk-derived proteins; and (30)
`aqueous solutions, alcohols (e.g., ethyl alcohol, propyl alco-
`sweetening, flavoring, coloring, perfuming and preservative
`hol, and benzyl alcohol), polyols (e.g., glycerol, propylene
`agents. Each such ingredient or material must be "accept-
`glycol, and polyethylene glycol), organic esters (e.g., ethyl
`able" in the sense of being compatible with the other
`oleate and tryglycerides), biodegradable polymers (e.g.,
`ingredients of the formulation and not injurious to the
`polylactide-polyglycolide, poly(orthoesters), and poly(an- 1
`5 subject. Ingredients and materials suitable for a selected
`hydrides)), elastomeric matrices, liposomes, microspheres,
`dosage form and intended route of administration are well
`oils (e.g., corn, germ, olive, castor, sesame, cottonseed, and
`known in the art, and acceptable ingredients and materials
`groundnut), cocoa butter, waxes, paraffins, silicones, talc,
`for a chosen dosage form and method of administration may
`silicylate, etc.
`be determined using ordinary skill in the art.
`
`Pharmaceutical compositions and nutritional supplements
`suitable for oral administration may be in the form of
`sition or nutritional supplement of the invention must be
`capsules, cachets, pills, tablets, powders, granules, a solution
`"acceptable" in the sense of being compatible with the other
`or a suspension in an aqueous or non-aqueous liquid, an
`ingredients of the formulation and not injurious to the
`oil-in-water or water-in-oil liquid emulsion, an elixir or
`subject. Carriers suitable for a selected dosage form and
`5 syrup, or a paste. These formulations may be prepared by
`intended route of administration are well known in the art, 2
`methods known in the art, e.g., by means of conventional
`and acceptable carriers for a chosen dosage form and
`pan-coating, mixing, granulation or lyophilization pro-
`method of administration can be determined using ordinary
`cesses.
`skill in the art.
`Solid dosage forms for oral administration (capsules,
`The pharmaceutical compositions and nutritional supple-
`tablets, pills, powders, granules and the like) may be pre-
`ments of the invention may, optionally, contain additional 30
`pared by mixing the active ingredient(s) with one or more
`ingredients and/or materials commonly used in pharmaceu-
`acceptable carriers and, optionally, one or more fillers,
`tical compositions and/or nutritional supplements. Such
`extenders, binders, humectants, disintegrating agents, solu-
`ingredients and materials include (1) fillers or extenders,
`tion retarding agents, absorption accelerators, wetting
`such as starches, lactose, sucrose, glucose, mannitol, and
`agents, absorbents, lubricants, and/or coloring agents. Solid
`silicic acid; (2) binders, such as carboxymethylcellulose, 3
`5 compositions of a similar type may be employed as fillers in
`alginates, gelatin, polyvinyl pyrrolidone, hydroxypropylm-
`soft and hard-filled gelatin capsules using a suitable excipi-
`ethyl cellulose, sucrose and acacia; (3) humectants, such as
`ent. A tablet may be made by compression or molding,
`glycerol; (4) disintegrating agents, such as agar-agar, cal-
`optionally with one or more accessory ingredients. Com-
`cium carbonate, potato or tapioca starch, alginic acid, certain
`pressed tablets may be prepared using a suitable binder,
`o lubricant, inert diluent, preservative, disintegrant, surface-
`silicates, sodium starch glycolate, cross-linked sodium car- 4
`active or dispersing agent. Molded tablets may be made by
`boxy methyl cellulose and sodium carbonate; (5) solution
`molding in a suitable machine. The tablets, and other solid
`retarding agents, such as paraffin; (6) absorption accelera-
`dosage forms, such as capsules, pills and granules, may
`tors, such as quaternary ammonium compounds; (7) wetting
`optionally be scored or prepared with coatings and shells,
`agents, such as cetyl alcohol and glycerol monosterate; (8)
`5 such as enteric coatings and other coatings well known in the
`absorbents, such as kaolin and bentonite clay; (9) lubricants, 4
`art. The tablets, and other solid dosage forms, may also be
`such as talc, calcium stearate, magnesium stearate, solid
`formulated so as to provide slow or controlled release of the
`polyethylene glycols, and sodium lauryl sulfate; (10) sus-
`active ingredient therein. They may be sterilized by, for
`pending agents, such as ethoxylated isostearyl alcohols,
`example, filtration through a bacteria-retaining filter. These
`polyoxyethylene sorbitol and sorbitan esters, microcrystal-
`compositions may also optionally contain opacifying agents
`line cellulose, aluminum metahydroxide, bentonite, agar- 5
`0 that release the active ingredient only, or preferentially, in a
`agar and tragacanth; (11) buffering agents; (12) excipients,
`certain portion of the gastrointestinal tract, optionally, in a
`such