WO 2006/0303552
`
`PCT/JP2005/005622
`
`- 20 -
`
`The obtained oils or fats (triglycerides) were
`acid).
`methylesterified, and the obtained fatty acid methyl
`ester mixture was analyzed by gas chromatography and
`found to have an arachidonic acid proportion of 40.84 wt%
`
`of the total fatty acid.
`The contents of palmitic acid, stearic acid, oleic
`linoleic acid, y-linolenic acid and dihomo-y-—
`acid,
`linolenic acid were 11.63%, 7.45%, 7.73%, 9.14%, 2.23%
`and 3.27% by weight, respectively.
`The arachidonic acid-
`containing oils or fats (triglycerides)
`(TGA40S) were
`also ethylesterified, and the fatty acid ethyl ester
`mixture including 40 wt% arachidonic acid ethyl ester was
`separated and purified by an established high-performance
`liquid chromatography method to obtain 99 wt% arachidonic
`acid ethyl ester.
`Example 2 Production of triglycerides including at
`least 5 mole percent 8A8
`After suspending 100 g of an ion-exchange resin
`carrier (Dowex MARATHON WBA: Dow Chemical)
`in 80 ml of
`Rhizopus delemar lipase aqueous solution (12.5% Talipase
`powder, Tanabe Pharmaceutical Co., Ltd.), 240 ml of cold
`acetone (-80°C) was stirred therewith and the mixture was
`dried under reduced pressure to obtain the immobilized
`
`lipase.
`Next, 80 gq of the triglycerides containing 40 wt%
`arachidonic acid (TGA40S) obtained in Example 1, 160 g of
`caprylic acid, 12 g of the aforementioned immobilized
`lipase and 4.8 ml of water were reacted for 48 hours at
`30°C while stirring (130 rpm). Upon completion of the
`reaction,
`the reaction solution was removed to obtain the
`activated immobilized enzyme.
`A 10 g portion of immobilized lipase (Rhizopus
`delemar lipase, carrier: Dowex MARATHON WBA) was then
`packed into a jacketed glass column (1.8 x 12.5 cm, 31.8
`mi volume), and the reaction oils or fats comprising a
`mixture of the TGA40S obtained in Example 1 and caprylic
`
`10
`
`15
`
`20
`
`25
`
`30
`
`35
`
`RIMFROST EXHIBIT 1024
`
`RIMFROST EXHIBIT 1024 page 2701
`
`page 2701
`
`

`

`WO 2006/0303552
`
`PCT/JP2005/005622
`
`- 21-
`
`acid (TGA40S: caprylic acid = 1:2) was flowed through the
`column at a fixed speed (4 ml/h) for continuous reaction,
`to obtain 400 g of reaction oils or fats.
`The coiumn
`temperature was 40-41°C.
`The unreacted caprylic acid and
`free fatty acids were removed from the obtained reaction
`oils or fats by molecular distillation, and then
`subjected to dietary oils or fats purification steps
`(degumming, deoxidation, deodorization, decolorizing) to
`obtain 8A8-containing oils or fats (triglycerides).
`The 8A8 proportion of the obtained 8A8-containing
`oils or fats (triglycerides) was determined by gas
`chromatography and high-performance liquid chromatography
`to be 31.6 mole percent.
`(Incidentally,
`the proportions
`of 8P8, 808, 8L8,
`8G8 and 8D8 were 0.6, 7.9, 15.1, 5.2
`and 4.8 mole percent, respectively.
`The fatty acids P,
`O, L, G and D bonded at the triglyceride 2-position
`
`represent palmitic acid, oleic acid,
`
`linoleic acid, y-
`
`linolenic acid and dihomo-y-linolenic acid, respectively,
`and therefore 8P8 represents 1,3-capryloyl-2-palmitolein-
`glycerol, 808 represents 1, 3-capryloyl-2-oleoyl-glycerol,
`8L8 represents 1,3-capryloyl-2-linoleoyl-glycerol,
`8G8
`represents 1,3-capryloyl-2-y-linolenoyl-glycerol and 8D8
`represents 1,3-capryloyl-2-dihomo-y-linolenoyl-glycerol).
`Separation and purification from the obtained 8A8-
`containing oils or fats (triglycerides) by an established
`high-performance liquid chromatography method yielded 96
`mole percent 8A8.
`Example 3 Evaluation of learning ability effect of
`TGA40S by Morris water maze learning test
`The experimental groups consisted of 56 two- to
`three-month-old male ICR mice, divided into a control
`diet group (27 mice) and a TGA40S-containing diet group
`(29 mice), with the control diet or TGA40S-containing
`diet shown in Table 1 being given to each group for 3
`weeks.
`Each group was further divided into non-
`restrained groups
`(non-restrained control diet group
`
`10
`
`15
`
`20
`
`25
`
`30
`
`35
`
`RIMFROST EXHIBIT 1024
`
`RIMFROST EXHIBIT 1024 page 2702
`
`page 2702
`
`

`

`WO 2006/0303552
`
`PCT/JP2005/005622
`
`—- 2? -
`
`(13), non-restrained arachidonic acid (ARA) diet group
`(15)) and restrained groups (restrained control diet
`group (14), restrained ARA diet group (14)).
`The
`restraining was accomplished using a wire mesh
`. restraining tube, once for a 6 hour period three weeks
`after the start of feeding.
`The control diet or TGA40S-—
`containing diet shown in Table 1 continued to be fed to
`each group for the remaining experiment period.
`The
`TGA40S used for the TGA40S-containing diet was the
`
`10
`
`product obtained in Example 1.
`
`Table 1 Experimental diet
`
`
`
`TY Centro diet
`200
`3 ee
`3
`150
`
`
`
`
`
`
`
`
`
`
`500
`
`-s0—t—C—~*S
`50
`50
`
`50eeee
`45
`
`35Po85
`35
`
`10 eeee
`10
`
`
`2
`Choline bitartrate
`
`0.05
`
`
`TGA40S
`
`
`
`0.05
`
`L5
`
`20
`
`25
`
`Since the daily ingestion was approximately 5 g per
`mouse,
`the daily intake of TGA40S was 25 mg per mouse.
`Also, since the total fatty acids bonded to the
`arachidonic acid-containing oils or fats (triglycerides)
`prepared in Example 1 included 40 wt% arachidonic acid,
`the daily intake of arachidonic acid was 10 mg per mouse.
`The 6-hour restraint with a wire mesh restraining
`tube was immediately followed by a Morris water maze
`learning test.
`The Morris water maze learning test is
`widely used in Europe and the U.S., and is conducted by
`pouring water blackened with India ink into a water tank
`(100 cm diameter, 35 cm height)
`(liquid surface height:
`20 cm), setting therein an escape platform of just a size
`to allow a mouse to stand (the escape platform is
`
`RIMFROST EXHIBIT 1024
`
`RIMFROST EXHIBIT 1024 page 2703
`
`page 2703
`
`

`

`WO 2006/0303552
`
`PCT/JP2005/005622
`
`-—~ 23 -
`
`submerged and invisible to a mouse swimming in the water
`tank), and then placing the mouse subject at a prescribed
`‘location of the water tank (starting point), forcing it
`to swim to the escape platform,
`in order to test its
`learning ability based on spatial recognition which is
`associated with the memory-governing hippocampus.
`
`The water temperature was 30°C +1°C, each trial was
`limited to 120 seconds with an interval of 60 seconds
`between trials, and five trials were conducted each day
`for 5 days.
`The time required for the mouse to reach the
`escape platform (escape latency time) was recorded as the
`learning index.
`No difference was observed between the
`control diet mice and ARA diet mice in the absence of
`restraint stress. However,
`the mice of the control diet
`group which had experienced restraint stress clearly
`exhibited reduced learning ability compared to the non-
`restrained mice, whereas mice given TAG40S (arachidonic
`acid) exhibited the same level of learning ability as the
`mice without restraint stress (Fig. 1).
`Thus,
`for the first time it has been clearly
`demonstrated that administration of TGA40S improves
`learning ability or cognitive ability which has declined
`as a result of stress, and that arachidonic acid exhibits
`an improving effect against decline in learning ability
`or cognitive ability as a result of stress.
`Example 4 Preparation of capsules comprising
`arachidonic acid-containing oils or fats (triglycerides)
`Water was added to 100 parts by weight of gelatin
`and 35 parts by weight of food additive grade glycerin
`for dissolution at 50-60°C,
`to prepare a gelatin coating
`with a viscosity of 2000 cp. Next, 0.05 wt% of vitamin 5
`oil was combined with the arachidonic acid-containing
`oils or fats (triglycerides) obtained in Example 1 to
`prepare filling 1. Vitamin E was also added to oils or
`fats (triglycerides) containing 32 mole percent of the
`8A8 obtained in Example 2 to prepare filling 2. Also, 50
`wt% of the arachidonic acid-containing oils or fats
`
`10
`
`15
`
`20
`
`25
`
`30
`
`35
`
`RIMFROST EXHIBIT 1024
`
`RIMFROST EXHIBIT 1024 page 2704
`
`page 2704
`
`

`

`WO 2006/0303552
`
`PCT/JP2005/005622
`
`-—- P4 -
`
`(triglycerides) obtained in Example 1 was combined with
`50 wt% fish oil
`(tuna oil:
`the eicosapentaenoic acid and
`docosahexaenoic acid proportions of the total fatty acids
`were 5.1% and 26.5%, respectively) and then 0.05 wt3
`vitamin E oil was added to prepare filling 3.
`Also, 80 wt% of the arachidonic acid-containing oils
`or fats (triglycerides) obtained in Example 1 was
`combined with 20 wt% fish oil
`(tuna oil: the
`
`eicosapentaenoic acid and docosahexaenoic acid
`proportions of the total fatty acids were 5.1% and 26.53%,
`respectively) and then 0.05 wt% vitamin E oil was added
`
`to prepare filling 4. Separately, 0.05 wt% of vitamin &E
`oil was combined with the 99% arachidonic acid ethyl
`ester obtained in Example 1 to prepare filling 5. These
`fillings 1 to 5 were used for production of soft capsules
`containing 180 mg of filling per capsule, obtained by
`capsule molding and drying by ordinary methods.
`Example 5 Use for oil infusion
`
`After combining 400 g of the oils or fats
`(triglycerides) containing 96 mole percent 8A8 obtained
`in Example 2, 48 g of purified egg yolk lecithin, 20 g of
`oleic acid, 100 g of glycerin and 40 ml of 0.1 N caustic
`
`soda and dispersing the mixture with a homogenizer,
`distilled water for injection was added to make 4 liters.
`This was emulsified with a high-pressure spray emulsifier
`to prepare a lipid emulsion.
`The lipid emulsion was
`dispensed into plastic bags at 200 ml per bag and then
`subjected to high-pressure steam sterilization treatment
`
`at 121°C for 20 minutes to prepare an oil infusion.
`Example 6 Use for juice
`
`A 2 g portion of B-cyclodextrin was added to 20 ml
`
`of 20% aqueous ethanol, and then 100 mg of the
`arachidonic acid-containing triglycerides obtained in
`Example 1
`(containing 0.05% vitamin E) were added thereto
`
`10
`
`15
`
`20
`
`25
`
`30
`
`35
`
`while stirring with a stirrer, and the mixture was
`
`incubated for 2 hours at 50°C. After room temperature
`
`RIMFROST EXHIBIT 1024
`
`RIMFROST EXHIBIT 1024 page 2705
`
`page 2705
`
`

`

`WO 2006/0303552
`
`PCT/JP2005/005622
`
`- 25 -
`
`cooling (approximately 1 hour), stirring was continued
`
`while incubating for 10 hours at 4°C. The resulting
`
`precipitate was recovered by centrifugal separation and
`
`then washed with n-hexane and lyophilized to obtain 1.8 g
`
`5
`
`of a cyclodextrin clathrate compound comprising
`
`arachidonic acid-containing triglycerides.
`
`A 1g portion
`
`of this powder was uniformly mixed into 10 L of juice to
`prepare a juice comprising arachidonic acid-containing
`
`triglycerides.
`
`RIMFROST EXHIBIT 1024
`
`RIMFROST EXHIBIT 1024 page 2706
`
`page 2706
`
`

`

`WO 2006/0303552
`
`PCT/JP2005/005622
`
`- 26 -
`
`CLAIMS
`
`A composition with a preventive or improvement
`1.
`effect on stress-induced brain function impairment and
`related symptoms or diseases, comprising arachidonic acid
`and/or a compound comprising arachidonic acid as a
`constituent fatty acid.
`2.
`A composition according to claim 1, wherein
`said compound comprising arachidonic acid as a
`constituent fatty acid is an arachidonic acid alcohol
`ester, or a triglyceride, phospholipid or glycolipid
`wherein all or a portion of the constituent fatty acid is
`arachidonic acid.
`
`A composition according to claim 2, wherein the
`3.
`triglyceride in which all or a portion of the constituent
`fatty acid is arachidonic acid is a triglyceride having
`medium chain fatty acids bonded at the 1,3-positions and
`arachidonic acid bonded at the 2-position.
`4,
`A composition according to claim 3, wherein
`Said medium chain fatty acids are selected from among Cé6-
`12 fatty acids.
`5.
`A composition with a preventive or improvement
`effect on stress-induced brain function impairment and
`related symptoms or diseases, comprising triglycerides
`which include a triglyceride in which all or a portion of
`the constituent fatty acid is arachidonic acid.
`6.
`A composition according to claim 5,
`characterized in that the arachidonic acid content of
`said triglycerides which include a triglyceride in which
`all or a portion of the constituent fatty acid is
`arachidonic acid,
`is at least 10 wt% of the total fatty
`acids of the triglycerides.
`7.
`A composition according to claim 5 or 6,
`wherein said triglycerides which include a triglyceride
`in which all or a portion of the constituent fatty acid
`is arachidonic acid, are extracted from a microorganism
`belonging to the genus Mortierella, Conidiobolus,
`Pythium, Phytophthora, Penicillium, Cladosporium, Mucor,
`
`10
`
`15
`
`20
`
`25
`
`30
`
`35
`
`RIMFROST EXHIBIT 1024
`
`RIMFROST EXHIBIT 1024 page 2707
`
`page 2707
`
`

`

`WO 2006/0303552
`
`PCT/JP2005/005622
`
`—_ 27 _
`
`Fusarium, Aspergillus, Rhodotorula, Entomophthora,
`
`Echinosporangium or Saprolegnia.
`
`8.
`
`A composition according to any one of claims 5
`
`te 7, wherein said triglycerides which include a
`triglyceride in which all or a portion of the constituent
`fatty acid is arachidonic acid, are triglycerides
`containing virtually no eicosapentaenoic acid.
`9.
`A composition with a preventive or improvement
`effect on stress-induced brain function impairment and
`
`related symptoms or diseases, comprising triglycerides of
`which at least 5 mole percent consists of a triglyceride
`having medium chain fatty acids bonded at the 1,3-
`positions and arachidonic acid bonded at the 2-position.
`10.
`A composition according to claim 9, wherein
`said medium chain fatty acids are selected from among C6é-
`12 fatty acids.
`
`A composition according to any one of claims 1
`ll.
`to 10, wherein said symptoms related to stress-induced
`brain function impairment include memory and learning
`ability impairment.
`
`
`
`A composition according to any one of claims 1
`12.
`to 10, wherein said symptoms related to stress-induced
`brain function impairment include cognitive ability
`impairment.
`
`13.
`
`A composition according to any one of claims 1
`
`to 10, wherein said symptoms related to stress-induced
`brain function impairment include depression.
`
`14.
`
`A composition according to any one of claims 1
`
`to 10, wherein said diseases related to stress-induced
`
`10
`
`15
`
`20
`
`25
`
`30
`
`brain function impairment include melancholia.
`
`A composition according to any one of claims 1
`15.
`to 14, wherein said composition is a food composition or
`pharmaceutical composition.
`
`A composition according to claim 15,
`16.
`characterized in that said food composition is a common
`food (food and drink), functional food, nutritional
`supplement,
`food for specified health uses, preterm
`
`35
`
`RIMFROST EXHIBIT 1024
`
`RIMFROST EXHIBIT 1024 page 2708
`
`page 2708
`
`

`

`WO 2006/0303552
`
`PCT/JP2005/005622
`
`- ?8 -
`
`infant formula,
`
`term infant formula,
`
`infant food,
`
`maternal food or geriatric food.
`17.
`A composition according to any one of claims 1
`to 16, which comprises docosahexaenoic acid and/or a
`compound comprising docosahexaenoic acid as a constituent
`fatty acid.
`18.
`A composition according to claim 17, wherein
`said compound comprising docosahexaenoic acid as a
`constituent fatty acid is a docosahexaenoic acid alcohol
`ester, or a triglyceride, phospholipid or glycolipid
`wherein all or a portion of the constituent fatty acid is
`
`docosahexaenoic acid.
`
`A composition according to claim 17 or 18,
`19.
`characterized in that the arachidonic
`
`in the
`acid/docosahexaenoic acid ratio (by weight)
`combination of said arachidonic acid and docosahexaenoic
`
`acid is in the range of 0.1 to 15.
`20.
`A composition according to any one of claims 1
`to 19, characterized in that the amount of
`eicosapentaenoic acid in the composition does not exceed
`1/5 of the arachidonic acid in the composition.
`21.
`A method for production of a dietary product
`having a preventive or improvement effect on stress—
`induced brain function impairment and related symptoms or
`diseases,
`the method being characterized by adding
`arachidonic acid and/or a compound comprising arachidonic
`acid as a constituent fatty acid alone, or in combination
`with a dietary material containing substantially no
`arachidonic acid or only a slight amount thereof.
`22.
`A method for prevention or medical treatment of
`stress-induced brain function impairment and related
`symptoms or diseases, which comprises administering
`arachidonic acid and/or a compound comprising arachidonic
`acid as a constituent fatty acid,
`to a patient in need of
`
`10
`
`15
`
`20
`
`25
`
`30
`
`35
`
`its administration.
`
`RIMFROST EXHIBIT 1024
`
`RIMFROST EXHIBIT 1024 page 2709
`
`page 2709
`
`

`

`WO 2006/0303552
`
`PCT/JP2005/005622
`
`
`
`
`
`dNO¥DL3TdTOULNODGSNIVULSSu-NON—e—
`
`
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`
`
`
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`
`
`
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`
`
`
`meie,
`
`|B14
`
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`
` 0001 ~0061
`“007
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`|0:09
`+008
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`6[$aD
`(SABp)IS3lYSVLFOV1d40AVGVv
`
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`
`[(dNOYDL3TGTOYLNOO-GSNIVULSTY“SA)10‘0>ds¥*“GO‘O>d:%“ASFUeOH)
`
`
`
`
`
`
`
`ESCAPE LATENCY TIME (sec)
`
`RIMFROST EXHIBIT 1024
`
`RIMFROST EXHIBIT 1024 page 2710
`
`page 2710
`
`
`
`
`

`

`Young, A
`
`-
`.
`.
`.
`"T" later document published after the internationalfiling date
`orpriority date and notin conflict with the application but
`cited to understand theprinciple or theory underlying the
`invention
`"X" documentof particular relevance; the claimed invention
`cannot be considered novel or cannot be considered to
`involve an inventive step when the documentis taken alone
`"¥" document of particular relevance; the claimed invention
`cannotbe consideredto involve an inventive step when the
`documentis combined with one or more other such docu-
`ments, such combination being obvious to a person skilled
`in the art.
`"&" document memberof the same patent family
`
`INTERNATIONAL SEARCH REPORT
`
`International Application No
`
`PCT/JP2005/005622
`
`+ CLASSIFICATION OF SUBJECT MATTER
`PC 7
`A61K31/202
`A61K31/232
`
`A23L1/30
`
`A61P25/00
`
`Accordingto International Patent Classification (IPC) or to both national classification and IPC
`B. FIELDS SEARCHED
`
`Minimum documentation searched (classification system followed byclassification symbols)
`IPC 7
`A61K A61P A23L
`
`Documentation searched other than minimum documeniation to the extent that such documentsare included in the fields searched
`
`Electronic data base consulted during the international search (name of data base and, wherepractical, search terms used)
`
`EPO-Internal, BIOSIS, EMBASE, WPI Data, PAd
`
`C. DOCUMENTS CONSIDERED TO BE RELEVANT
`
`
`
`Category °|Citation of document, with indication, where appropriate, of the relevant passages Relevantto claim No.
`
`1-22
`
`WO 2004/028529 A (SUNTORY LIMITED;
`AKIMOTO, KENGO; KOGA, YOSHIHIKO)
`8 April 2004 (2004-04-08)
`claims 1-35
`examples 1-8
`
`EP 1 419 768 A (SUNTORY LIMITED)
`19 May 2004 (2004-05-19)
`page 9,
`lines 25-32
`examples 1-8
`
`WO 02/19839 A (UNIVERSITY OF MARYLAND
`BIOTECHNOLOGY INSTITUTE)
`14 March 2002 (2002-03-14)
`claims 1,2
`
`Further documents are listed in the continuation of box C.
`
`Patent family membersarelisted in annex.
`
`Date of mailing of the international search report
`
`20/07/2005
`Authorizedofficer
`
`° Specia} categories of cited documents:
`
`"A" document defining the general state of the art whichis not
`consideredto beof particular relevance
`"E" earlier documentbut published on orafter the international
`filing date
`"L" document which may throw doubtson priority, claims)or
`which
`is cited
`to establish the publicationdate of another
`citation or other special reason (as specified)
`“O" documentreferring to an oral disclosure, use, exhibition or
`other means
`"P" document published prior to the international filing date but
`later than the priority date claime:
`Date of the actual completion of the international search
`
`29 June 2005
`
`Nameand mailing address of the ISA
`European Patent Office, P.B. 5818 Patentlaan 2
`NL — 2280 HV Rijswijk
`Tel. (431-70) 340-2040, Tx. 31 651 epo ni,
`Fax: (+31-70) 340-3016
`
`Form PCT/ISA/210 (second sheet) (January 2004)
`
`RIMFROST EXHIBIT 1024
`
`RIMFROST EXHIBIT 1024 page 2711
`
`page 2711
`
`

`

`INTERNATIONAL SEARCH REPORT
`
`International Application No
`PCT/JP2005/005622
`
`C.(Continuation) DOCUMENTS CONSIDERED TO BE RELEVANT
`
`
`
`Category °|Citation of document,with indication, where appropriate, of the relevant passages Relevantto claim No.
`
`"PREVENTION OF
`AUGUSTE L-J ET AL:
`STRESS-INDUCED EROSIVE GASTRITIS BY
`PARENTERAL ADMINISTRATION OF ARACHIDONIC
`ACID"
`JOURNAL OF PARENTERAL AND ENTERAL
`NUTRITION,
`vol. 14, no. 6, 1990, pages 615-617,
`XP009049858
`ISSN: 0148-6071
`
`abstract
`
`Form PCT/ISA/210 (continuation of second sheet) (January 2004)
`
`RIMFRO ST EXHIBIT 1024
`
`RIMFROST EXHIBIT 1024 page 2712
`
`page 9712
`
`

`

`INTERNATIONAL SEARCH REPORT
`
`
`
`
`International application No.
`
`PCT/JP2005/005622
`
`Box Il Observations where certain claims were found unsearchable (Continuation of item 2 of first sheet)
`
`This Internationa! Search Report has not been established in respect of certain claims underArticle 17(2)(a) for the following reasons:
`
`4.
`
`
`
`
`
`2, [| Claims Nos.:
`becausetheyrelate to parts of the International Application that do not comply with the prescribed requirements to such
`an extent that no meaningful International Search can be carried out, specifically:
`
`3. [| Claims Nos.:
`
`because they are dependentclaims and are not drafted in accordance with the second andthird sentences of Rule 6.4(a).
`
`Box Ill Observations where unity of invention is lacking (Continuation of item 3 of first sheet)
`
`This International Searching Authority found multiple inventions in this international application, as follows:
`
`
`Claims Nos.:
`because they relate to subject matter not required to be searched bythis Authority, namely:
`Although claim 22 is directed to a method of treatment of the human/anima]
`body,
`the search has been carried out and based on the alleged effects of the
`compound/composition.
`
`
`
`
`
`
`
`
`
` Asall required additional search fees weretimely paid bythe applicant, this International Search Report coversall
`
`
` As all searchable claims could be searched without effort justifying an additional fee, this Authority did not invite payment
`of any additional fee.
`
`
`3. [| As only someof the required additional search fees were timely paid by the applicant, this International Search Report
`covers only those claimsfor which fees were paid, specifically claims Nos.:
`
`searchable claims.
`
`
`
`
` No required additional search fees were timely paid by the applicant. Consequently, this international Search Report is
`
`restricted to the Invention first mentioned in the claims; it is covered by claims Nos.:
`
`
`Remark on Protest
`[] The additional search fees were accompanied by the applicant's protest.
`
`[| No protest accompanied the paymentof additional search fees.
`RIMFROST EXHIBIT 1024 page 2713
`
`Form PCT/ISA/210 (continuation of first sheet (2)) (January 2004)
`
`RIMFROST EXHIBIT 1024
`
`page 2713
`
`

`

`INTERNATIONAL SEARCH REPORT
`Information on patent family members
`
`Patent document
`cited in search report
`
`Publication
`date
`
`Patent family
`member(s)
`
`International Application No
`PCT/JP2005/005622
`Publication
`date
`
`WO 2004028529
`
`A
`
`08-04-2004
`
`EP 1419768
`
`A
`
`19-05-2004
`
`WO 0219839
`
`A
`
`14-03-2002
`
`2003267818
`2499902
`1542670
`2004028529
`
`2003048831
`2456049
`1419768
`2004266874
`1561206
`03013497
`
`9258601
`2424570
`1324671
`0219839
`2002110582
`
`19-04-2004
`08-04-2004
`22-06-2005
`08-04-2004
`
`21-02-2003
`20-02-2003
`19-05-2004
`30-12-2004
`05-01-2005
`20-02-2003
`
`22-03-2002
`14-03-2002
`09-07-2003
`14-03-2002
`15-08-2002
`
`
`Form PCT/ISA/210 (patent family annex) (January 2004)
`
`page 2714
`
`RIMFROST EXHIBIT 1024
`
`RIMFROST EXHIBIT 1024 page 2714
`
`

`

`(12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY(PCT)
`
`(19) World Intellectual Property
`Organization
`International Bureau
`
`
`
`(43) International Publication Date
`28 April 2005 (28.04.2005)
`
`(10) International Publication Number
`WO 2005/037848 A2
`
`(51)
`
`(21)
`
`International Patent Classification’:
`
`CO07F9/10
`
`(74)
`
`International Application Number:
`PCT/IL2004/000957
`
`(81)
`
`(22)
`
`International Filing Date: 21 October 2004 (21.10.2004)
`
`(25)
`
`Filing Language:
`
`(26)
`
`Publication Language:
`
`English
`
`English
`
`(30)
`
`Priority Data:
`158552
`
`22 October 2003 (22.10.2003)
`
`IL
`
`(84)
`
`(71)
`
`(72)
`(75)
`
`Applicant (for all designated States except US): ENZY-
`MOTEC LTD.
`[ILAL]; Ramat-Gavriel Industrial Park,
`P.O. Box 6, 23106 Migdal HaEmek (IL).
`
`Inventors; and
`Inventors/Applicants (for US only): BEN DROR,
`Gai [ILAL]; P.C. Box 126, 30835 Moshav Ofer (IL).
`PLATT, Dorit [ILAL]; P.O. Box 249, 17906 Shimshit
`(IL). FARKASH, Orly [IL/IL]; P.O. Box 217, 17906
`Shimshit UL). ZUABI, Rassan [ITLAIL]; 19320 Kfar Neen
`(IL). BAR-ON, Zohar [IL/IL]; 32 Yodfat Street, 21950
`Karmiel (IL). SHULMAN,Avidor[IL/IL]; 29 HaGomeh
`Street, 36090 Kiryat Tivon (L). PELLED, Dori [ILAL];
`29 HaShahar Street, 45325 Hod Hasharon (IL).
`
`Agents: LUZZATTO,Kfir ct al.; P.O. Box 5352, 84152
`Beer Sheva (IL}.
`
`Designated States (unless otherwise indicated, for every
`kind of national protection available): AE, AG, AL, AM,
`AT, AU, AZ, BA, BB, BG, BR, BW, BY, BZ, CA, CH, CN,
`CO, CR, CU, CZ, DE, DK, DM, DZ, EC, EE, EG, ES, FI,
`GB, GD, GE, GH, GM, HR, HU,ID, IL, IN,IS, JP, KE,
`KG, KP, KR, KZ, LC, LK, LR, LS, LT, LU, LV, MA, MD,
`MG, MK, MN, MW, MX, MZ, NA, NI, NO, NZ, OM,PG,
`PH, PL, PT, RO, RU, SC, SD, SE, SG, SK, SL, SY, TJ, TM,
`TN,TR, TT, TZ, UA, UG, US, UZ, VC, VN, YU, ZA, ZM,
`ZW.
`
`Designated States (unless otherwise indicated, for every
`kind of regional protection available): ARIPO (BW, GH,
`GM, KE, LS, MW, MZ, NA, SD, SL, SZ, TZ, UG, ZM,
`ZW), Eurasian (AM, AZ, BY, KG, KZ, MD, RU, TJ, T™),
`European(AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, FI,
`FR, GB, GR, HU,IE, IT, LU, MC, NL, PL, PT, RO,SE, SI,
`SK, TR), OAPI (BF, BJ, CF, CG, CI, CM, GA, GN, GQ,
`GW, ML, MR, NE, SN, TD, TG).
`
`Published:
`
`without international search report and to be republished
`upon receipt of that report
`
`For two-letter codes and other abbreviations, refer to the "Guid-
`ance Notes on Codes and Abbreviations" appearing at the begin-
`ning of each regular issue of the PCT Gazette.
`
`(54) Title: LIPIDS CONTAINING OMEGA-3 AND OMEGA-6 FATTY ACIDS
`
`(57) Abstract: Disclosed is a lipid preparation comprising a glycerophospholipid or salt, conjugate and derivatives thereof, particu-
`larly phosphatidylserine (PS), phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidyl-inositol (PD, phosphatidyl]-
`glycerol (PG) and phosphatidic acid (PA), and poly-unsaturated fatty acid (PUFA) acyl groups, particularly long-chain poly-unsat-
`urated fatty acid (LC-PUFA)acyl groups such as omega-3 and/or omega-6 acyl groups, wherein said PUFA is covalently bound to
`said glycerophospholipid. The disclosed preparations possess an improved bioactivity, and are useful in the treatment of various
`cognitive and mental conditions and disorders and for maintenance of normal functions of brain-related systems and processes.
`
`RIMFROST EXHIBIT 1024
`
`RIMFROST EXHIBIT 1024 page 2715
`
`page 2715
`
`
`
`2005/037848A2/IINIMIAININAINAININNATMIMMANERTAAA
`
`WwO
`
`

`

`WO 2005/037848
`
`PCT/1IL2004/000957
`
`LIPIDS CONTAINING OMEGA-3 AND OMEGA-6 FATTY ACIDS
`
`]
`
`Field of the Invention
`
`The present
`
`invention relates
`
`to. phospholipids
`
`and polar
`
`lipids
`
`preparations which are enriched with omega-3 and/or omega-6 fatty acids
`
`covalently attached to the lipid backbone. The phospholipid preparations
`
`of the invention are particularly useful as nutraceuticals, food additives
`
`and/or pharmaceutical agents for the treatment of various conditions, in
`
`particular related to cognitive functions.
`
`Background of the Invention
`
`and
`lipids, nitrogen containing lipids,
`Lipids, and especially polar
`carbohydrate containing lipids (phospholipids, sphingosines, glycolipids,
`ceramides,
`sphingomyelins) are the major building blocks of’ cell
`
`membranes, tissues, etc. Additionally they play important roles in signal
`
`transduction processes and in a variety of biochemical and biosynthetic
`
`pathways.
`
`Glycerophospholipids, lipids based on a glycerol backbone and containing
`
`a phosphate head group, are the main building blocks of cell membranes.
`
`Since most, if not all, biochemical processes involve cell membranes, the
`
`structural and physical properties of membranes in different tissues is
`
`crucial
`
`to the normal and efficient functioning of membranes in all
`
`biochemical processes.
`
`In light of the emerging functional foods category in the area of dietary
`
`lipids many health benefits have been attributed to the consumption of
`
`certain fatty acids. For example, it has been reported in many research
`
`studies that polyunsaturated fatty acids (PUFA) of the type omega-3 and
`
`omega-6, have several health benefits on cardiovascular disease, immune
`
`disorders and inflammation, renal disorders, allergies, diabetes, and
`
`RIMFROST EXHIBIT 1024
`
`RIMFROST EXHIBIT 1024 page 2716
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`

`

`WO 2005/037848
`
`PCT/IL2004/000957
`
`2
`
`cancer. These types of fatty acids are naturally occurring mainly in fish
`
`and algae, where they are randomly distributed on the sn-1, sn-2, and sn-3
`
`positions of the glycerol backbone oftriglycerides.
`
`The professional literature emphasizes the importance of an adequatediet
`
`containing omega-3 fatty acids. Extensive clinical studies investigating the
`
`importance of Docosahexaenoic acid (DHA), one of the most important
`
`omega-3 fatty acids,
`
`in the brain, found that low levels of DHA are
`
`associated with depression, memory loss, dementia, and visual problems.
`
`All studies showed a dramatic improvement in the elderly brain function
`
`as blood levels of DHA increased.
`
`Other known benefits of DHA include:
`
`lower risk of arrhythmias,
`
`reduction in the risk of sudden cardiac death, lower plasma triglyceride
`
`levels and reduced blood clotting tendency. Furthermore, DHA may have
`
`importance in the field of brain functioning enhancement, baby formula
`
`fortification, diabetics and cancer. Nutritional studies, investigating the
`
`importance of DHA in the brain, found that low levels of DHA are
`
`associated with depression, memory loss, cognitive impairment, dementia
`
`and visual problems.
`
`The human body does not adequately synthesize DHA. Therefore it is
`
`necessary to obtain it from the diet. Humans obtain DHA from their diets,
`
`initially through the placenta, then from breast milk, and later through
`
`dietary sources, such as fish, red meats, animal organ meats and eggs.
`
`Popular fish like tuna, salmon and sardines are rich sources. Until
`
`recently, the primary source of DHA dietary supplements has been fish
`
`oils. The ability of enzymes to produce the omega-6 and omega-3 family of
`
`products of linoleic and alpha-linolenic acid declines with age. Because
`
`DHA synthesis declines with age, as we get older our need to acquire DHA
`
`directly from diet or supplements increases. In fact, several recent
`
`RIMFROST EXHIBIT 1024
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`RIMFROST EXHIBIT 1024 page 2717
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`WO 2005/037848
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`PCT/IL2004/000957
`
`publications suggested DHA to be considered as essential fatty acid [for
`
`example, Muskiet, F. et al. (2004) J Nutr. 184(1):188-6].
`
`3
`
`Because DHA is important for signal transmission in the brain, eye and
`
`nervous system, many consumers concerned with maintaining mental
`
`acuity are searching for a pure, safe way to supplement their DHA levels.
`
`Polyunsaturated acids, in particular long chain, such as omega-38 and 6,
`
`have been shown to confer many valuable health benefits on the
`
`population. The global market for long-chain PUFAs, including the food
`
`segment, is rapidly growing.
`
`The majority of efforts in the industry are however invested in the
`
`improvement of PUFA processing techniques andin the creation of higher
`
`concentrated grades of PUFA derivatives
`
`to accommodate dietary
`
`supplements and functional foods needs.
`
`The academic and industrial communities are less concerned regarding
`
`the evaluation of different delivery approaches of PUFA in order to
`
`enhance their bio-availability and their efficacy in term of their known
`
`variety of health benefits. These benefits range from prevention and
`
`treatment of CVD, diabetes, cognitive disorders and/or decline, visual
`
`disorders, skin conditions, learning disorders, etc. Additionally, PUFAs
`
`have been shown to assist in the cognitive and visual development of
`
`infants.
`
`PUFA-linids
`
`PS-PUFA
`
`Phosphatidylserine, also known as PS, is a natural phospholipid with bio-
`
`functionality that has made it one of the most promising dietary
`
`supplements in the field of brain nutrition. PS and its health benefits have
`
`RIMFROST EXHIBIT 1024
`
`RIMFROST EXHIBIT 1024 page 2718
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`WO 2005/037848
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`PCT/IL2004/000957
`
`4
`
`been known to the scientific and. nutrition communities since the 1970's.
`
`Numerous studies have been conducted in order to establish this efficacy
`in a variety of cognitive and mental functions. Those studies have shown
`
`fight early stages of
`fight dementia,
`that PS can improve memory,
`Alzheimer's disease, reduce stress and tension, improve attention span,
`enhance mood andfight depression, to name but few.
`
`PS is one of the most important building blocks of cell membranes in the
`
`brain. Hence, the level of PS in brain cell membranes ensuresthefluidity
`
`and. structure of these membranes. The normal level ensures normal and
`
`efficient signal transduction processes, e