Patent Application: US-57855104-A

Abstract:
methods of controlling serum glucose levels in an individual are described , the methods including the step of administering to said individual a therapeutic food composition comprising a waxy and / or hydrothermally treated starch . the method may be used to treat or prevent hypoglycaemia in patients susceptible to hypoglycaemic episodes , for example patients with glycogen storage disease , diabetes or liver disease . the method may also be used in sports nutrition . also described are compositions for use in the methods .

Description:
as described above , the present inventors have discovered that existing treatments for conditions characterised by hypoglycaemic episodes may be improved and / or supplemented by the use of waxy starches as sources of α - glucan , thus enabling significant improvement to control over the rate of glucose formation and appearance in the blood mammals . such starches significantly outperform the conventionally used ‘ corn starch ’ ( native maize starch ) in terms of duration of glucose release due to amylase hydrolysis in the small intestine . moreover , the inventors have shown that the glucose release profile may be further dramatically prolonged by modifications to the optimised starch e . g . by hydrothermal treatment for example , by heat moisture treatment . indeed , hydrothermal treatment also provides considerable improvement in conventional non - waxy starches . thus , the invention also extends to the methods of the first , second and third aspect of the invention , wherein the waxy starch is substituted by any hydrothermally treated starch , preferably heat moisture treated starch ( whether waxy or non - waxy ). starches are produced by plants as roughly spherical granules ranging in diameter from & lt ; 5 to & gt ; 50 μm . depending on source they contain ˜ 11 - 17 % moisture , ˜ 82 - 88 % α - glucan , & lt ;˜ 1 . 5 % lipid and & lt ;˜ 0 . 6 % protein . the α - glucan comprises two types of molecules : amylose and amylopectin . the former is an essentially linear molecule comprising about 99 % α -( 1 - 4 ) and about 1 % α -( 1 - 6 ) bonds with a molecular weight of ˜ 500 , 000 . amylopectin is much bigger than amylose with a molecular weight of a few million and is heavily branched with ˜ 95 % α -( 1 - 4 ) and ˜ 5 % α -( 1 - 6 ) bonds . the exterior chains of amylopectin associate together as double helices which themselves register together to form crystalline laminates . these crystalline laminates are interspersed with amorphous material comprising non - crystalline ( branched regions ) of amylopectin plus amylose . the amylose may form inclusion complexes in cereal starches with lipids causing the presence of two forms of the molecule : lipid complexed and lipid free . in normal starches , amylopectin is the ‘ seat ’, of crystallinity . waxy starches have a greater proportion of crystallinity due to the higher amylopectin content . high amylose starches contain both amylopectin and amylose generated crystalline material . starches containing & lt ;˜ 20 % amylose ( 80 % amylopectin ) are commonly referred to as ‘ waxy ’, ˜ 20 - 40 % are commonly referred to as ‘ normal ’ and ˜& gt ; 40 % are commonly referred to as high amylose or amylo - starches . normal maize and wheat starches are , for example , ˜ 30 % amylose . the semi - crystalline native starch granules are insoluble and largely indigestible by man &# 39 ; s digestive enzymes . the control of native starch digestion in man is not well understood although it does not provide a major nutritional focus as most starches are processed prior to cooking . processing of starch incorporates cooking in water which disrupts the crystalline regions and ‘ gelatinises ’ the starch . gelatinised starches are very digestible because of their amorphous nature by amylases and related enzymes in the small intestine of man . native and resistant starches ( see below ), although in part digested in the small intestine , are fermented in the colon . products of carbohydrate fermentation in the colon include short chain fatty acids ( scfas ) and gasses like carbon dioxide , hydrogen and methane . resistant starch takes a number of forms and simply resists hydrolysis by enzymes synthesised in the small intestine of man . this includes : small food particles entrapping starch ; native starch ; recrystallised ( retrograded ) starch and ; chemically modified starch . if starches are hydrolysed ( typically chemically with acids or enzymatically with α - amylase and amyloglucosidase ) smaller molecules called ‘ dextrins ’ are generated . products may be as small as the smallest possible monosaccharide glucose or be slightly hydrolysed but still polymeric . glucose syrups are made from starch hydrolysis and contain variable proportions of sugars and dextrins depending on the nature and extent of conversion . the extent of conversion is usually defined as dextrose equivalence ( de ) which equates reducing power of the hydrolysate to that of pure dextrose ( glucose ). maltodextrins are dp20 or less , gras quality , tasteless and very soluble . they are easily digestible and are used in energy drinks because of their solubility and reportedly relatively slow digestibility compared to glucose ( which is simply absorbed ). the difference in rate of glucose appearance in the blood as a consequence of drinking glucose or maltodextrin solutions is relatively small ( e . g . ˜ 45 minutes ) because of the extent of conversion of the maltodextrin . in the present invention , any suitable semi - crystalline or crystalline starch may be used . in preferred embodiments , the starch of and for use in the invention is a waxy starch . the starch may be a naturally produced starch or may be synthetically produced using any suitable method e . g . plant breeding or biotechnological methods ( including transgenic technology etc .). preferred native starches are waxy with an average diameter of approximately 15 - 35 μm . as discussed above and shown in the examples below , the inventors have found that particularly good results are obtained when using hydrothermally treated starch . two main methods are currently used for the hydrothermal treatment of starch : heat - moisture treatment ( high temperature , low moisture ) and annealing ( high moisture , low temperature ). heat and moisture treated starch is typically produced by exposing moist starch ( e . g . 15 - 30 % moisture ) to temperatures of e . g . 95 ° c . to 130 ° for periods up to 30 hours ( typically 16 - 24 ). these ranges do not exclude other heat - moisture profiles . for example , hmt starch for use in the invention may be produced by thermally treating starch in a sealed container under the following conditions : 20 % moisture and 105 ° c . for 16 hours . the treated starch may then be cooled to room temperature , air - dried and then passed through 300 um sieve . such heat moisture treatment results in a number of significant property changes to starches . the extent of the effect varies with the type of starch but in general the effects are : increased gelatinisation temperature reduced water absorption and swelling power changed x - ray diffraction pattern increased enzyme susceptibility as described herein , although heat moisture treatment results in starches having increased susceptibility to enzymatic degradation , the inventors have surprisingly shown that when used in methods of the invention , heat moisture treated starches provide significantly greater prolongation of the time period over which serum glucose levels are maintained compared to the corresponding non heat moisture treated starches . in certain embodiments of the invention the starch of and for use in the invention is annealing treated starch . any suitable annealing treated starch may be used . annealing is a process in which starch granules are treated for a relatively long time in excess amounts of water at a temperature slightly higher then room temperature . typically , annealing of starch involves incubation of starch granules in water (& gt ; 40 % w / w ), for a time period in the range 1 hour to 10 days at a temperature between the glass transition and the gelatinisation temperature . preferred annealing conditions are less than 10 ° c . below the onset of gelatinisation temperature , in excess water for up to 7 days . “ treatment ” ( which , unless the context demands otherwise , is used interchangeably with “ therapy ”, includes any regime that can benefit a human or non - human animal . the treatment may be in respect of an existing condition or may be prophylactic ( preventative treatment ). treatment may include curative , alleviation or prophylactic effects . the invention extends to a therapeutic food composition for the treatment of diseases characterised by hypoglycaemic episodes , wherein said composition comprises a semi - crystalline starch . the therapeutic food compositions of and for use in the present invention may consist solely of said starches or preferably may comprise further additives . such additives may contribute merely to the palatability of the composition , e . g . flavourings , or may contribute significant calorific value , for example , sugars with a more rapid release profile than the starches , or lipids . these compounds may be incorporated to slow gastric emptying and facilitate the effect ( e . g . amino acids , lipids etc .). the therapeutic food composition can take a variety of forms , for example as a food , a food supplement , a liquid , an emulsion or mixture thereof . preferably , it is prepared as a ready to eat foodstuff , for example as a snackbar , a baked product , pasta or drink . alternatively , the therapeutic food composition may be administered as a pharmaceutical composition , which will generally comprise a suitable pharmaceutical excipient , diluent or carrier selected dependent on the intended route of administration . some suitable routes of administration include ( but are not limited to ) oral , rectal or parenteral ( including subcutaneous , intramuscular , intravenous , intradermal ) administration . for intravenous injection the active ingredient will be in the form of a parenterally acceptable aqueous solution which is pyrogen - free and has suitable ph , isotonicity and stability . those of relevant skill in the art are well able to prepare suitable solutions using , for example , isotonic vehicles such as sodium chloride injection , ringer &# 39 ; s injection , lactated ringer &# 39 ; s injection . preservatives , stabilisers , buffers , antioxidants and / or other additives may be included , as required . however , the composition is preferably for administration orally . pharmaceutical compositions for oral administration may be in tablet , capsule , powder or liquid form . a tablet may comprise a solid carrier such as gelatin or an adjuvant . liquid pharmaceutical compositions generally comprise a liquid carrier such as water , petroleum , animal or vegetable oils , mineral oil or synthetic oil . physiological saline solution , dextrose or other saccharide solution or glycols such as ethylene glycol , propylene glycol or polyethylene glycol may be included . examples of the techniques and protocols mentioned above and other techniques and protocols which may be used in accordance with the invention can be found in remington &# 39 ; s pharmaceutical sciences , 16th edition , oslo , a . ( ed ), 1980 . the therapeutic food compositions of and for use in the invention are preferably administered to an individual in a “ therapeutically effective amounts ”, this being sufficient to show benefit to the individual . the actual amount administered , and rate and time - course of administration , will depend on the nature and severity of what is being treated . prescription of treatment , e . g . decisions on dosage etc , is ultimately within the responsibility and at the discretion of general practitioners and other medical doctors , and typically takes account of the disorder to be treated , the condition of the individual patient , the site of delivery , the method of administration and other factors known to practitioners . the optimal dose can be determined by physicians based on a number of parameters including , for example , age , sex , weight , severity of the condition being treated , the active ingredient being administered and the route of administration . the invention will now be described further in the following non - limiting examples . reference is made to the accompanying drawings in which : fig2 shows a comparison of the hydrolysis profile of native starches using the karkalas et al ( 1992 ) procedure ; fig3 shows blood glucose level after consumption of native starches ; fig4 shows a comparison of the blood lactate level after consumption of native starches ; fig5 shows a comparison of blood glucose after consumption of two native starches ( wheat and waxy maize ) with added pregelatinised ( maize ) starch ; fig6 shows a comparison of the blood lactate level after consumption of two native starches ( wheat and waxy maize ) with added pregelatinised ( maize ) starch ; fig7 shows a comparison of blood glucose after consumption of starch ( native waxy maize and soluble ) encapsulated with pectin and alginate . fig8 shows a comparison of blood lactate after consumption of starch ( native waxy maize and soluble ) encapsulated with pectin or alginate . fig9 shows a comparison of blood glucose after consumption of starch ( native waxy maize , soluble ) encapsulated with lipid . fig1 shows a comparison of blood glucose after consumption of heat - moisture treated waxy maize starch , waxy maize and normal maize starch . fig1 shows a comparison of blood lactate after consumption of heat - moisture treated waxy maize starch , waxy maize and normal maize starch . fig1 shows a comparison of digestibility of native and heat - moisture treated waxy maize starches . fig1 shows a comparison of digestibility of native and heat - moisture treated normal maize starches . common native starches ( barley , maize , potato , rice and wheat ) were evaluated using the karkalas et al ( 1992 ) ( in vitro ) method to identify their amylase hydrolysis profile and potential for slow release of energy in individuals . these data are presented in fig2 . as can be seen from fig2 that rice starch has a fast energy release profile initially followed by a much slower process . in contrast , potato and high amylose starches show great resistance towards amylase hydrolysis and are nearly untouched by the enzyme . starches from normal maize , waxy maize and wheat show continuous slow release energy profile . these data provide the basis for an in vitro selection of the most appropriate starch for this purpose ( as discussed later ). note that they do not define the rate or extent of hydrolysis in the actual gut but provide a means of ordering the rate of extent of hydrolysis based on the in vitro system . under clinical supervision , individuals suffering from gsd were fed 60 g samples of native starches dispersed in semi - skimmed milk . the amount of blood glucose and lactate were monitored and are presented in fig3 and 4 . native potato starch was not consumed in view of is resistance to digestion ( and cause of potential colonic disturbance accordingly ). these data show that waxy rice starch released glucose very quickly where the highest ( too high ) initial glucose peak ( 8 . 7 mmoll − 1 ) at 1 hour post ingestion was obtained . the blood glucose level then dropped to 3 mmoll − 1 within 4 . 5 hours ( 270 minutes ). normal rice showed a much lower initial glucose peak with a longer release profile corresponding to 3 . 2 mmoll − 1 within 5 hours ( 300 minutes ) but less glucose released in the time course of the experiment compared to the waxy rice starch . high amylose starch too extensively restricted glucose release ( although this could be moderated by physical / chemical / enzymatic or biotechnological modification ). the normal maize starch (‘ corn starch ’) exhibited a low glucose peak 1 hour ( 6 . 6 mmoll − 1 ) after ingestion with an extended release of 2 . 9 mmoll − 1 after 300 minutes . the waxy maize starch surprisingly showed the optimal release profile with less than 7 mmoll − 1 blood glucose 1 hour post ingestion , a significant glucose release profile for up to 6 hours ( 330 minutes ) which dropped to 2 . 9 mmoll − 1 at this point . lactate in the blood also reflected the starch consumed ( fig4 ). the high amylose maize starch provided the least lactate response ( highest lactate ) as it was little digested ( fig3 ). the greatest reduction in lactate was achieved by the waxy maize starch and in common with the previous data promotes its potential use for gsd and similar conditions requiring slow release of energy . based on these data , there is clearly a granule size and compositional effect that regulates native starch hydrolysis to glucose in the gut . there is a balance between choosing a starch for therapy based on the 1 hour glucose peak , duration of release and the amount ( integrated area ) of glucose release with time . a preferred starch for the purpose , therefore : a ) is highly crystalline ( semi - crystalline ) with waxy starches providing the most appropriate crystalline ( amylopectin ) matrices for this purpose . b ) has reasonably large granules without exceeding the digestive capacity . rice starches (˜ 5 μm diameter on average ) are too small . maize starch granules are preferred (˜ 20 - 25 μm diameter on average ). it is recognised that the cereal starches contain lipid and that other starches may be more appropriate in terms of size and composition . however , in view of the lack of digestibility and potential dangers of eating large granules ( which may cause colonic lesions ) it is proposed that granules in excess of ˜ 40 μm diameter are not consumed for this purpose . digestion of native starches in the presence of a pre - gelatinised starch thickener under clinical supervision , individuals suffering from gsd were fed 60 g samples of two native starches ( wheat or waxy maize ), each sample containing 54 g of either starch and 6 g pregelatinised maize starch ( national b37 , national starch & amp ; chemical ) dispersed in cold semi - skimmed milk . the amount of blood glucose and lactate were monitored and are presented in fig5 and 6 . these data show that even in the presence of amorphous ( pre - gelatinised ) starch the waxy maize starch resists digestion ( fig5 ) more than the wheat starch , which contains a bi - modal distribution of small (˜ 10 μm average diameter ) and large (˜ 25 μm average diameter ) granules but with similar composition ( amylose , lipid , moisture and protein ). this is reflected in a lower blood lactate ( even though the patients started with a higher lactate content when presented with the waxy maize starch ( as shown in fig6 ). the importance of this work is that it shows that even if the waxy starch is mixed with other materials that have the capacity to provide a quicker glucose response it can still provide a slow release function . native waxy maize starch ( amioca powder t , national starch ) was encapsulated in soluble starch ( crystal tex 626 , national starch ) and pectin ( lm - 104as - fs , cpkelco ) or alginic acid ( manugel gmb , manugel ) according to tester and karkalas ( 1999 ). the final starch to non - starch polysaccharide ( nsp ) ratio was 5 . 7 : 1 or 19 : 1 . the proportion of the soluble starch to native starch varied according to the proportion of native starch used for the two conditions but was the same for both non - starch polysaccharide conditions and simply serves as a comparison . under clinical supervision , individuals suffering from gsd were fed 70 g or 63 g ( depends on the starch to nsp ratio ) samples of nsp encapsulated starch dispersed in cold semi - skimmed milk . the amount of blood glucose and lactate were monitored and are presented in fig7 and 8 . these data show that , although the amount of starch modifies the extent of glucose release as expected , the alginate or pectin components do not stretch out the release profile much beyond 5 hours ( 300 minutes ). hence , the presence of a non - starch polysaccharide ‘ raft ’ or food matrix is not enough in itself to slow the rate of starch hydrolysis accordingly ( whether native or soluble ). the blood lactate response reflects the blood glucose where alginate appears to reduce lactate production more markedly than pectin ( since it restricts hydrolysis more ). starch ( amioca powder t , national starch ) with or without addition of soluble starch ( crystal tex 626 , national starch ) was encapsulated in lipid ( revel a , loders croklaan b . v .) as follows . the lipid was dissolved in the minimal amount of ethanol possible to dissolve the starch . the starch was then thoroughly mixed with the ethanol solution until homogeneous . the starch was laid on a tray and air at 35 ° c . was allowed to flow over the ethanol / lipid / starch system ( in a fume cupboard ) until the ethanol had all evaporated from the system . the final starch to lipid ratio was 9 : 1 . when used , the proportion of soluble starch was 10 % of the total starch fraction . under clinical supervision , individuals suffering from gsd were fed 66 g samples of lipid encapsulated starch dispersed in cold semi - skimmed milk . the amount of blood glucose was monitored and is presented in fig9 . these data show that the lipid restricts the amount of starch digestion at all times ( see previous figures for comparison ). overall , this approach is not appropriate for the control of glucose release ( extent of hydrolysis ) from the starch as the amount released over time and the actual duration is reduced . starch ( amioca powder t , national starch ) was thermally treated in a sealed container under the following conditions : 20 % moisture and 105 ° c . for 16 hours . the treated starches were then cooled to room temperature , air - dried and then passed through 300 μm sieve . under clinical supervision , individuals suffering from gsd were fed 60 g or 90 g samples of heat - moisture treated starch dispersed in cold semi - skimmed milk . the amount of blood glucose and lactate were monitored and are presented in fig1 and 11 . ( i ) heat moisture treated ( hmt ) waxy maize starch has a much reduced initial glucose response at 60 minutes than native waxy maize starch ( fig1 ). ( ii ) because of the reduced initial response more can be fed to be within acceptable levels of glucose increase at this time ( where a preferred response is & lt ; 8 mmol l − 1 ). ( iii ) as a consequence of the above , greater amounts could be fed ( 90 g versus 60 g ) leading to 7 . 5 hour ( 450 minutes ) profile where the hmt starch can still maintain the blood glucose at ˜ 2 . 5 mmol l − 1 . ( iv ) the glucose response provides an acceptable and desirable lactate response accordingly ( fig1 ). similar results were obtained when repeating the experiments on further patients ( results not shown ). these data are reinforced by the in vitro assay as shown in fig1 . here the hmt treatment can be shown to clearly restrict the hydrolysis of the waxy maize starch . hence , the combination of a waxy starch and its heat moisture treatment allows for the formation of a desirable slow release of glucose therapy . the waxy maize starch is potentially more crystalline than normal or high amylose starches in view of the high amylopectin content . a particularly preferred type of starch for this purpose is : semi crystalline with , preferably , the highest proportion of crystallinity possible and with amylase accessibility enhanced by the heat moisture processing . moreover , in order to show that the advantages conferred by hydrothermal treatment is not limited to waxy starches , the digestibility of native and heat - moisture treated normal maize starch was tested using the same assay as in fig1 . the results are shown in fig1 . as shown in fig1 , hydrothermal treatment of normal maize starch ( i . e . non - waxy starch ) improves the hydrolysis profile of the starch . thus , the results support the use of hydrothermally treated normal starch for slow release glucose therapy in the methods of the invention . all documents referred to in this specification are herein incorporated by reference . various modifications and variations to the described embodiments of the inventions will be apparent to those skilled in the art without departing from the scope and spirit of the invention . although the invention has been described in connection with specific preferred embodiments , it should be understood that the invention as claimed should not be unduly limited to such specific embodiments . indeed , various modifications of the described modes of carrying out the invention which are obvious to those skilled in the art are intended to be covered by the present invention . berggren , a ., johansson , m . l ., larsson , k ., lindberg , a - m . and wiklander , j . 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