Note: Descriptions are shown in the official language in which they were submitted.
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. 1
LIQUID DETERGENTS CONTAINING PROTEOLYTIC
ENZYME, PEPTIDE ALDEHYDE AND CALCIUM IONS
-
TECHNICAL FIELD
This invention relates to liquid detergent compositions cont~ining en_ymes.
10 More specifically, this invention pertains to liquid d~ r~elll compositions
cont~ining a detersive ~ulr~ l, a proteolytic enzyme, a peptide aldehyde, and
calcium ions. The combination of peptide aldehyde and calcium ions act to provide
synergistic protease inhibitor benefits.
BACKGROUND OF THE INVENTION
Protease-co~ g Iiquid aqueous detergents are well-known, especially in
the context of laundry washing. A commonly encountered problem in such
protease-cont~ining liquid aqueous detergents is the degradation phenomenon by the
proteolytic enzyme of second enzymes in the composition, such as amylase, lipase,
and cellulase, or on the protease itself. As a result, the stability of the second
20 enzyme or the protease itself in the detergent composition is affected and the
detergent composition consequently performs less well.
In response to this problem, it has been proposed to use various protease
inhibitors or stabilizers. For inct?nre, various references have proposed the use of
the following compounds to aid in the stabilization of enzymes: bçn7~midine
25 hydrochloride, lower aliphatic alcohols or carboxylic acids, mixtures of a polyol and
a boron compound, aromatic borate esters, and calcium, particularly calcium
formate. Recently, it was discovered that certain peptide aldehydes act to stabilize
protease enzyme.
Although these compounds have been used to varying success in liquid
30 detergents, they are not free of problems. For example peptide aldehydes are rather
ex~ si~/e and create complexities for the fonnnl~tors, especially for liquid
d~enl~. Other inhibitors such as calcium and boric acids are less expensive but
do not stabilize enzymes ~ well as peptide aldehydes. It is thus an object of the
present invention to provide a protease inhibitor system which is economical,
35 effective and suitable for use in a liquid detergent composition.
~ n ~ onse to this object, the present invention proposes to use a combination
of calcium ions and peptide aldehydes as reversible protease inhibitors in aqueous
~ ... .. . .. .
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liquid detergent compositions. The presence of both calcium and peptide aldehydeprovides a synergistic stabilization of the protease. This novel combination provides
the formulator added flexibility in cle~igning a stabilization system. The levels of
peptide aldehyde and calcium can be adjusted to deliver the most cost effective
formula and to minimi7e product stability problems that o~en arise from the
presence of divalent ions in a liquid detergent matrix.
In particular, the present invention allows for the use of very low levels of
peptide aldehydes in the liquid detergent compositions herein. This is particularly
critical in the formulation of relatively inexpensive, concentrated liquid detergent
10 compositions which are encompassed by the present invention.
Because the combination of calcium and peptide aldehydes are so efficient in
inhibiting proteases, another advantage of the present invention is that even enzymes
which are highly sensitive to proteolytic degradation can now be incorporated inliquid detergent compositions comprising a protease. Moreover, it has also been
15 discovered that the increased stability of the protease enzyme allows for improved
skincare benefits. These benefits include softening of the skin and hands and less
drying from exposure of the hands to the dishwashing liquor.
BACKGROUND ART
It has been proposed to use various protease inhibitors or stabilizers. For
20 instance, US 4,566,985 proposes to use benzamidine hydrochloride; EP 376 705
proposes to use lower aliphatic alcohols or carboxylic acids; EP 381 262 proposes to
use a mixture of a polyol and a boron compound; and EP91870072.5 proposes to usearomatic borate esters. See also U.S. Pat. No. 5,030,378 issued July 9, 1991. Also
see US4,261,868; US4,404,115; US4,318,818; and EP130,756.
The use of peptide derivatives for the inhibition of proteins appears to have
been disclosed in therapeutic applications. EP 293 881 discloses the use of peptide
boronic acids as inhibitors of trypsin-like serine proteases. EP 185 390 and US
4,399,065 disclose the use of certain peptide aldehydes derivatives for the inhibition
of blood coagulation. J 90029670 discloses the use of optically active alpha amino
30 aldehydes for the inhibition of enzymes in general. See also "Inhibition of
Thrombin and Trypsin by Tripeptide Aldehydes", Int. J. Peptide Protein Res., Vol12 (1978), pp. 217-221; Gaal, Bacsy & Rappay, and "Tripeptide Aldehyde Protease
Inhibitors May Depress in Vitro Prolactin and Growth Hormone Release"
Endocrinolo~y, Vol. 116, No. 4 (1985), pp. 1426-1432; Rappay, Makara, Bajusz &
35 Nagy. Certain peptide aldehydes have also been disclosed in EP-A-473 502 for
inhibiting protease-me~ tecl skin irritation.
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In particular see EP185,390, WO94/04651, published 3 March 1994,
W094/04652, published 3 March 1994, EP 583,536, published February 23, 1994,
EP 583,535, published February 3, 1994, EP 583,534, published February 23, 1994,WO 93/13125, published July 8, 1993, US4,529,525, US4,537,706, US4,537,707,
5 and US5,527,487.
SUMMARY OF THE INVENTION
The invention herein is a liquid detergent composition comprising:
a) an effective amount of a detersive surfactant;
b) an active proteolytic enzyme;
10 c) a source of calcium ions; and
d) a peptide aldehyde having the formula:
Z-B-NH-CH(R)-C(O)H
wherein B is a peptide chain comprising from 1 to 5 amino acid moieties; Z is
an N-capping moiety selected from the group consisting of phosphoramidate
[(R"O)2(O)P-3, sulfenamide [(SR")2-], sulfonamide [(R"(O)2S-], sulfonic acid
[SO3H], phosphinamide [(R")2(O)P-], sulfamoyl derivative [R"O(O)2S-], thiourea
[(R")2N(O)C-], thiocarbamate [R"O(S)C-], phosphonate [R"-P(O)OH],
amidophosphate [R"O(OH)(O)P-], carbamate (R"O(O)C-), and urea (R"NH(O)C-),
wherein each R" is independently selected from the group consisting of straight or
branched Cl-C6 llncubstituted alkyl, phenyl, C7-Cg alkylaryl, and cycloalkyl
moieties, wherein the cycloalkyl ring may span C4-Cg and may contain one or moreheteroatoms selected from the group consisting of O,N,and S (preferred R" is
selected from the group con.~icting of methyl, ethyl, and benzyl); and R is selected
from the group con.cicting of straight or branched C1 - C6 unsubstituted alkyl,
phenyl, and C7 - Cg alkylaryl moieties.
Without being limited by theory, it is believed that the combined source of
calcium ion and peptide aldehyde provides more than additive stability to the
proteolytic enzyme.
Preferably, the liquid detergent compositions herein comprise, by weight of
composition:
a) from about I to about 9S%, preferably from about 8% to about 70%, of said
detersive surfactant;
b) from about 0.0001% to about 5%, preferably from about 0.0003% to about
0.1 %, of an active proteolytic enzyme;
c) from about 0.00001% to about 5%, preferably from about 0.0001% to about
1 %, more preferably from about 0.0006% to about 0.5%, of a peptide aldehyde as
described hereinbefore; and
.
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d) from about 0.01% to about 1%, preferably from about 0.05% to about 0.5%, of
calcium ion.
The proteolytic enzyme useful herein is preferably a subtilisin-type protease
and may be selected from the group consisting of Alcalase~), Subtilisin BPN',
Protease A, Protease B, and mixtures thereof.
The source of calcium ion for use herein is preferably selected from calcium
formate, calcium xylene sulfonate, calcium chloride, calcium acetate, calcium
sulfate. and mixtures thereof.
The dishcare compositions herein may contain further detersive adjuncts,
including but not limited to, one or more of the following: suds boosters, chelants,
polyacrylate polymers, dispersing agents, dyes, perfumes, processing aids, and
mixtures thereof. Moreover for dishcare compositions, the liquid detergent
compositions may further comprise an effective amount of amylase enzyme.
Additionally, the dishcare compositions may optionally comprise an effective
amount of a source of boric acid and a diol. Typically dishcare compositions will
optionally, but preferably, comprise from about 0.25% to about 10%, preferably
from about 0.5% to about 5%, more preferably from about 0.75% to about 3%, by
weight of boric acid or a compound capable of forming boric acid and a diol, e.g.
1 ,2-propaneidiol .
In a preferred embodiment for heavy duty detergent compositions useful in
laundry care, the liquid detergent composition further comprises an effective amount
one or more of the following enzymes: lipase, amylase, cellulase, and mixtures
thereof. Preferably for laundry compositions, the second enzyme is lipase and isobtained by cloning the gene from Humicola Lanuginos~ and ~x~lc;sslllg the gene in
Aspergillus Oryzae. Lipase is utilized in an amount of from about 10 to about 18000
lipase units per gram, preferably from about from about 60 to about 6000 units per
gram.
In another pl~rell~d composition useful for laundry care, the second enzyme is
a cellulase derived from Humicola Insolens and is utilized in an amount of from
about 0.0001% to about 0.1% by weight of the total composition of said cellulase.
The compositions herein may contain further detersive adjuncts, including but
not limited to, one or more of the following: suds boosters, builders, soil release
polymers, polyacrylate polymers, dispersing agents, dye transfer inhibitors, dyes,
perfumes, processing aids, brighteners, and mixtures thereof. Additionally, for
laundrycare compositions, the detersive surfactant is typically present in an amount
of from about 10% to about 70%, by weight of total composition. Moreover, the
laundry compositions may optionally comprise an effective amount of a source of
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boric acid and a diol. Typically laundry compositions will optionally, but
preferably, comprise from about 0.25% to about 10%, preferably from about 0.5% to
about 5%, more preferably from about 0.75% to about 3%, by weight of boric acid
or a compound capable of forming boric acid and a diol, e.g. 1,2-propaneidiol.
All percentages and proportions herein are by weight, and all references cited
are hereby incorporated by reference, unless otherwise specifically indicated.
DETAILED DESCRIPTION OF THE INVENTION
Definitions - The present detergent compositions comprise an "effective
amount" or a "stain removal-improving amount" of individual components defined
herein. An "effective arnount" or "stain removal-improving amount" is any amountcapable of measurably improving soil cleaning or stain removal from a substrate,i.e., soiled fabric or soiled dishware, when it is washed by the consumer. In general,
this amount may vary quite widely.
By "synergy" or "more than additive" as used herein is meant that the enzyme
stability benefit when the calcium and peptide aldehydes are combined is greaterthan the sum of the individual benefits obtained when only one of the components is
present in a detergent composition.
The liquid aqueous detergent compositions according to the present invention
comprise four essential ingredients: (A) a peptide aldehyde or a mixture thereof, (B)
a proteolytic enzyme or a mixture thereof, (C) a detersive surfactant, and (D)
calcium ion. The compositions according to the present invention preferably further
comprise (E) a detergent-compatible second enzyme or a mixture thereof, and may
further comprise (F) other optional ingredients.
PeDtide aldeh~des - The detergent compositions according to the present
invention comprise, as a first essential ingredient, a peptide aldehyde having the
forrnula:
Z-B-NH-CH(R)-C(O)H
wherein B is a peptide chain colllpllsing from 1 to 5 amino acid moieties; Z
is an N-capping moiety selected from the group concicting of phosphorarnidate
[(R"0)2(0)P-], sulf~on~mi~le [(SR")2-], sulfonamide [(R"(0)2S-], sulfonic acid
[S03H], phosphin~mide [(R")2(0)P-], s--lf~mc yl derivative [R"0(0)2S-], thiourea[(R")2N(O)C-], thioc~banlate [R"O(S)C-], phosphonate [R"-P(O)OH],
arnidophosphate [R"O(OH)(O)P-], carbamate (R"O(O)C-), and urea (R"NH(O)C-),
wherein each R" is indep~nflently selected from the group consisting of straight or
branched C I -C6 unsubstituted alkyl, phenyl, C7-Cg alkylaryl, and cycloalkyl
moieties, wherein the cycloalkyl ring may span C4-Cg and may contain one or moreheteroatoms selected from the group concicting of O,N,and S (preferred R" is
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selected from the group con~icting of methyl, ethyl, and benzyl); and R is selected
from the group consisting of straight or branched C I - C6 unsubstituted alkyl,
phenyl, _nd C7 - Cg alkylaryl moieties.
Preferred R moieties are selected from the group consisting of methyl, iso-
propyl, sec-butyl, iso-butyl, -C6Hs, -CH2-C6Hs, and -CH2CH2-C6Hs, which
respectively may be derived from the _mino acids Ala, Val, Ile, Leu, PGly
(phenylglycine), Phe, and HPhe (homophenylAIAnine) by converting the carboxylic
acid group to an aldehyde group. While such moieties are therefore not amino acids
(_nd they may or may not have been synthPci7Pcl from an amino acid precursor), for
purposes of simplification of the exemplification of inhibitors useful here, thealdehyde portion of the inhibitors are indicated as derived from amino acids by the
addition of "H" after the A~alogous amino acid [e.g., "-Ala~" represents the
chemical moiety "-NHCH(CH3)C(O)H"].
Preferred B peptide chains are selected from the group consisting of peptide
chains having the amino acid sequences according to the general formula:
Z-A5-A4-A3-A2-A I -NH-CH(R)-C(O)H
such that the following amino acids, when present, are:
Al is selected from Ala, Gly;
A2, if present, is selected from Val, Ala, Gly, Ile;
A3, if present, is selected from Phe, Leu, Val, Ile;
A4, if present, is any amino acid, but preferably is selected from Gly, Ala;
A5, if present, is any amino acid, but preferably is Gly, Ala, Lys.
The present invention aldehydes may be prepared from the corresponding
amino acid whereby the C-tf rrninAI end of said amino acid is converted from a
carboxylic group to ~ aldehyde group. Such aldehydes may be prepared by ~nown
processes, for instance as described in US 5015627, EP 185 930, EP 583,534, and
DE3200812.
While not wanting to be bound by theory it is believed that the peptide
aldehydes according to the present invention bind to the proteolytic enzyme in the
liquid detergent composition, thereby inhibiting said proteolytic enzyme. Upon
dilution in water, the proteolytic activity is restored by dissociation of the proteolytic
enzyme/peptide aldehyde complex.
The N-tenninAl end of said protease inhibitors according to the present
invention is protected by one of the N-capping moiety protecting groups selectedfrom the group con~i~ting of c~bA.nAtPs, ureas, sulfon~mi-lPs,
phosphonamides,thioureas, sulfenamides, sulfonic acids, phosphinAmides,
thiocarbAm~tes, amidophosphates, and phosphonamides. However, in a highly
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plefelled embodiment of the present invention, the N-terminal end of said protease
inhibitor is protected by a methyl, ethyl or benzyl carbamate [CH30-(O)C-;
CH3CH20-(O)C-; or C6HsCH20-(O)C-], methyl, ethyl or benzyl urea [CH3NH-
(O)C-; CH3CH2NH-(O)C-; or C6HsCH2NH-(O)C-], methyl, ethyl or benzyl
sulfonamide [CH3S02-; CH3CH2S02-; or C6HsCH2S02-], and methyl, ethyl or
benzyl amidophosphate ICH30(0H)(O)P-; CH3CH20(0H)(O)P-; or
C6HsCH20(0H)(O)P-] groups.
Synthesis of N-capping groups can be found in the following references:
Protective Groups in Or~anic Chemistry. Greene, T., Wuts, P., John Wiley & Sons,New York, 1991, pp 309-405; March, J, Advanced Or~anic ChemistrY, Wiley
Interscience, 1985, pp. 445, 469, Carey, F. Sundberg, R., Advanced Or~anic
Chemistr~, Part B, Plenum Press, New York, 1990, pp. 686-89; Atherton, E.,
Sheppard, R., Solid Phase Peptide Svnthesis, Pierce Chemical, 1989, pp. 3-4; Grant,
G., Svnthetic Peptides, W. H. Freeman & Co. 1992, pp. 77-103; Stewart, J., Young,
1 s J., Solid Phase Peptide SYnthesis. 2nd Edition, IRL Press, 1984, pp. 3,5,11,14- 18,
28-29. Bodansky, M., Principles of Peptide Synthesis, Springer-Verlag, 1988, pp.62, 203, 59-69; Bodansky, M., Peptide Chemistry, Springer-Verlag, 1988, pp. 74-81,
Bodansky, M., Bodansky, A., The Practice of Peptide Synthesis, Springer-Verlag,
1984, pp. 9-32.
Examples of peptide aldehydes for use herein are: CH3S02Phe-Gly-Ala-Leu-
H, CH3S02Val-Ala-Leu-H, C6HsCH20(0H)(O)P-Val-Ala-Leu-H, CH3CH2S02-
Phe-Gly-Ala-Leu-H, C6HsCH2S02-Val-Ala-Leu-H, C6HsCH20(0H)(O)P-Leu-
Ala-Leu-H, C6HsCH20(0H)(O)P-Phe-Ala-Leu-H, and CH30(0H)(O)P-Leu-Gly-
Ala-Leu-H.
In the Synthesis Examples hereinafter methods are disclosed to synthesi7~o
certain of these peptide aldehydes.
Synthesis Example 1
Synthesis of the te~lal)el)tide aldehyde Moc-Ala-Phe-Gly-Ala-LeuH
(a) Ala-Leu-OMe.HCL: To a solution of 3.0 g (14.83 mrnol) Ala-Leu-OH, which is
dissolved in 50 ml of MeOH and cooled to 0~C, is added 2.43 ml (33.36 mmol)
thionyl chloride dropwise. This solution is stirred overnight at room temperature and
evaporated to dryness providing quantitative recovery of the desired product.
(b) Cbz-Gly-Ala-Leucine methyl ester: To a solution of 0.414 g (1.98 mmol) Cbz-
Gly-OH and 0.500 g (1.98 mmol) Ala-Leu-OMe.HCl in CH2C12 is add 0.607 ml
TEA followed immediately by 0.355 ml DEPC. The solution is stirred overnight,
evaporated, and the residue partitioned between EtOAc and lN HCI. The organic
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phase is washed successively with saturated NaE~C03 and saturated NaCl, dried
(MgSO4)and evaporated to afford 0.650 g of pure product.
(c) Moc-Ala-Phe-OH: To a solution of 1.0 g (4.23 mrnol) Ala-Phe which is
dissolved in 4.23 ml IN NaOH and cooled to 0~C, 0.419 g (4.44 mmol) is aded
methyl chloroformate dropwise. At the same time, in a separate addition funnel, an
additional 4.23 ml IN NaOH is added such that the pH is m~int~ined between 9.0-
9.5. After addition is complete the reaction is stirred 30 minutes at 0~C and 2 h at
room temperature. At this point the solution is cooled to 0~ and the pH adjusted to
0 9.5. This basic solution is washed with EtOAc (lX, 100 ml). The aqueous (0~C) is
then adjusted to pH = 2.5 (2N HCl) and extracted with EtOAc (3X, 50 ml), dried
(MgSO4) and evaporated to provide 1.07 g pure product.
(d) Moc-Ala-Phe-Gly-Ala-Leu-OMe: To a solution of 0.500 g (1.22 mrnol) Cbz-
Gly-Ala-Leucine methyl ester in 10 ml MeOH is added 0.100 g 10% Pd/C. This
solution is hydrogenated in the presence of 0.600 ml 4.0M HCI/Dioxane (under
balloon pressure) for 1 h, filtered through celite and evaporated. This residue is
suspended in CH2C12, 0.342 ml (2.45 mmol) TEA is added followed by 0.359 g
(1.22 mmol) Moc-Ala-Phe-OH and 0.219 ml (1.34 mmol) DEPC. After stirring
overnight the solvent is evaporated, the residue partitioned between EtOAc and IN
HCI and washed successively with saturated NaHCO3 and NaCI. Drying,
evaporation and column chromatography yield 0.450 g of the pure product.
(e) Moc-Ala-Phe-Gly-Ala-Leucinol: A solution is prepared by dissolving 0.182 g
(1.64 munol) CaC12 in a mixture of 4 ml ethanol and 2 ml THF. This mixture is
cooled to -15~C and 0.450 g (0.820 mrnol) Moc-Ala-Phe-Gly-Ala-Leu-OMe is
added followed by 0.124 g (3.28 mrnol) NaBH4. The reaction is stirred for 2 h and
q~en~he~l with 10 ml lN HCI. The solvents are evaporated and the rçm~ining
aqueous layer partitioned with EtOAc. The organic phase is then washed with
saturated NaHCO3 and saturated NaCI. Drying (MgSO4), evaporation and
chromatography affords 0.256 g of pure product.
(f) Moc-Ala-Phe-Gly-Ala-LeuH: A solution is prepared by adding 0.623 g (1.47
mmol) Dess-Martin periodinane to 1.8 L CH2Cl2 followed by stirring for 10
minlltes This solution is then cooled to 0~C and 0.256 g (0.490 mmol) Etoc-Phe-
Gly-Ala-Leucinol is added in one portion. The reaction is continued for 2 h and
poured into a solution con.ci~ting of 2.55 g (10.47 mmol) Na2S2O3 in 30 ml
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saturated NaHCO3. After stirring for 10 minutes the mixture is extracted with
EtOAc (2X, 50 ml). The combined extracts are dried (MgSO4), evaporated. and
chromatographed on silica to provide 0.125 g of pure product.
SYnthesis Example 2:
SYnthesis of the tripeptide aldehyde Etoc-Phe-Gly-Ala-LeuH
(a) Ala-Leu-OMe.HCL: To a solution of 450 g (2.20 mol) Ala-Leu-OH, which is
dissolved in 4.5 L of MeOH and cooled to 0~C, is added 178.6 ml (4.95 mol) of
thionyl chloride dropwise. The solution is stirred overnight at room tenl~e,d~ lre and
evaporated to dryness providing 543 g (97.1 % yield) of the desired product to be
used as is.
(b) Etoc-Phe-Gly-OH: To a solution of 450 g (2.03 mol) Phe-Gly which is
dissolved in 2026 ml IN NaOH and cooled to 0~C, is added methyl chloroformate
(3. l ml, 40.0 mmol) dropwise. At the same time, in a separate addition funnel, an
additional 2026 ml IN NaOH is added such that the pH is m~int~ine-l between 9.0-9.5. After addition is complete the reaction is stirred 30 minutes at 0~C and 2 h at
room temperature. At this point the solution is cooled to 0~ and the pH adjusted to
9.5. This basic solution is washed with EtOAc (IX, 4 L). The aqueous (0~C) is then
adjusted to pH = 2.5 (2N HCI) and extracted with EtOAc (3X, 8L), dried (MgSO4),
filtered, and the solvent removed to afford 546 g (91.3% yield) pure product.
(c) Etoc-Phe-Gly-Ala-Leu-OMe: To a solution of 470 g (1.86 mol) Etoc-Phe-Gly-
OH and 546 g (1.86 mol) Ala-Leu-OMe.HCl in 8 liters CH2CI2 570 ml (4.09 mol)
TEA is added followed by 310.4 ml (2.046 mol) DEPC. After stirring overnight thesolvent is evaporated and replaced with EtOAc (4 L). This solution is washed
con~ec~l~ively with 2 liters each of 2N HCI, sat'd NaHCO3 and sat'd NaCI. The
organic phase is then dried (MgSO4), filtered and evaporated to yield 916 g (93%yield) of the desired m~t~
(d) Etoc-Phe-Gly-Ala-Leucinol: To a solution of 45.10 g (0.406 mol) CaC12 in l Lethanol and I L THF 100 g (0.203 mol) of Etoc-Phe-Gly-Ala-Leu-OMe is added
and the mixture cooled to -15~C. To this solution 30.7 g (0.812 mmol) NaBH4 is
carefully added followed by stirring for 2 h. Subsequently the reaction is quenched
with 100ml 0.1N HCI. This solution is transfered to 4 L of IN HCI and extracted
with EtOAc (3X, 2.75 L). The combined EtOAc layers are washed with 4 L
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saturated NaHC03, dried (MgSO4) and evaporated. Trituration (twice) with ether (4
L) provides 69.2 g (73.4% yield) of the product.
(e) Etoc-Phe-Gly-Ala-LeuH: A solution is prepared by adding 165.4 g (0.39 mol)
Dess-Martin periodinane to 1.8 L CH2C12 followed by stirring for 10 minutes. This
solution is then cooled to 0~C and 60 g (0.13 mol) Etoc-Phe-Gly-Ala-Leucinol
added in one portion. The reaction is continued for 105 minutes and poured into a
solution consisting of 6 L H2O, 393 g NaHCO3 and 431.7 g (1.74 mol) Na2S2O3.
After stirring for 10 minutes the phases are separated and 2 additional extractions
10 (1.5 L each) with CH2C12 are ~c~rulllled. The combined extracts are dried (MgSO4),
evaporated, and triturated with (2X, I L) ether to provide 51.7 g (86.2% yield) of the
product.
Synthesis Exarnple 3:
Svnthesis of the dipeptide aldehyde Moc-GlY-Ala-LeuH
(a) Ala-Leu-OMe.HCL: To a solution of 3.0 g (14.83 mmol) Ala-Leu-OH, which is
dissolved in 50 ml of MeOH and cooled to 0~C, is added 2.43 ml (33.36 mrnol) of
thionyl chloride dropwise. The solution is stirred overnight at room temperature and
evaporated to dryness providing a quantitative yield of the desired product.
(b) Cbz-Gly-Ala-Leucine methyl ester: To a solution of 0.414 g (1.98 rnrnol) Cbz-
Gly-OH and 0.500 g (1.98 mmol) Ala-Leu-OMe.HCI in CH2C12 0.607 ml TEA is
added followed im mer~i~tely by 0.355 mi DEPC. The solution is stirred overnightand then evaporated. The residue is partitioned between EtOAc and lN HCI, the
25 organic phase is washed with saturated NaHCO3 and saturated NaCI, dried
(MgS04) and evaporated providing 650 mg of pure product.
(c) Moc-Gly-Ala-Leucine methyl ester: To a solution of 2.0 g (4.90 mmol) Cbz-
Gly-Ala-Leucine methyl ester which is dissolved in 20 ml MeOH is added 0.200 g
30 10% Pd/C. This is hydrogenated in the presence of 2.45 ml (9.81 mmol) 4.0M
HCI/Dioxane for 2 h after which the reaction is thoroughly ou~g~c~ed and filtered
through Celite to remove the catalyst. Evaporation of the MeOH affords 1.45 g ofpure product which is suspended in 45 ml CH2C12 and cooled to 0~C. To this
solution 1.45 ml (3.25 mmol) TEA is added followed by 0.362 ml methyl
35 chloroformate. After stirring overnight the CH2C12 is evaporated and the residue
partitioned between EtOAc and 1 N HCI. The organic phase is separated and washed
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WO 98/13459 PCTIUS97/16622
11
sequentially with NaHCO3 and NaCl. Drying, (MgS04), evaporation and
chromatographic purification affords 0.~20 g of desired product.
(d) Moc-Gly-Ala-Leucinol: To a solution of 0.168 g (1.51 mmol) CaC12 in 25 ml
ethanol and 15 ml THF is added 0.250 g Moc-Gly-Ala-Leucine methyl ester. This
solution is cooled to -15~C and 0.114 g (3.02 mmol) NaBH4 is added in one portion.
After stirring 2 h the reaction is quenched with 20 ml lN HCI, concentrated on
rotovape and extracted with EtOAc (2x 50ml). The combined extracts are washed
with saturated NaHCO3 and NaCI, dried (MgSO4) and evaporated. Purification on
0 silica provides 0.167 g of the pure product.
(e) Moc-Gly-Ala-LeuH- A solution is prepared by adding 0.418 g (0.989 rnmol)
Dess-Martin periodinane to 5 ml CH2C12 followed by stirring for 10 minutes. Next0.100 g (0.330 mmol) Moc-Gly-Ala-Leucinol is added in one portion and the
15 reaction stirred for 2 h and poured into a 25 ml solution of saturated NaHCO3cont~ining 1.72 g (6.93 mmol) Na2S2O3. After stirring an additional 10 minutes the
solution is extracted with EtOAc (3X, 50 ml), dried (MgSO4) and evaporated.
Chromatography on silica affords 0.016 g of the desired product.
Svnthesis Example 4:
Synthesis of N-(methylsulfonyl)-Phe-Gly-Ala-LeuH
(a) N-Ms-Phe-Gly-OH: To a solution of 2.0 g (9.0 mmol) Phe-Gly-OH, which is
dissolved in 9 ml lN NaOH and cooled to 0~C, is added simultaneously 0.766 ml (
9.9 mmol) of methane sulfonyl chloride and 9 ml lN NaOH, in separate addition
25 funnels. After addition is complete the reaction is stirred 15 minlltes at 0~C and 1 h
at room temperature. At this point the solution is cooled to 0~C, the pH adjusted to
9.5 and is washed with EtOAc (lX, 50 m!). The aqueous phase (0~C) is then
adjusted to pH = 2.5 (2N HCI) and extracted with EtOAc (3X, 50 ml), dried
(MgSO4), filtered, and the solvent removed to afford 2.0 g pure product.
(b) N-Ms-Phe-Gly-Ala-Leucinol: A solution of is prepared by dissolving 0.500 g
(1.67 mmol) N-Ms-Phe-Gly-OH in 15 ml THF, cooling to -15~C, and adding 0.366
ml (3.33 mmol) NMM followed by 0.216 ml ( 1.67 mmol) isobutyl chloroformate.
This solution is stirred 5 minutes and 0.374 g (1.67 mmol) Ala-Leucinol.HCI, in a
35 mixture of 10 ml THF and minim~l DMF, are added. Stirring is continued at 0~C for
15 minutes and 2 h at room teln~ldlllre. The solution is quenched with S ml lN
HCl, extracted with EtOAc (3X, 50 ml), the combined extracts are washed with sat'd
CA 02266~27 1999-03-23
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12
NaHCO3 and sat'd NaCI. The resulting organic phase is then dried (MgSO4),
filtered, evaporated and chromatographed on silica to yield 0.260 g of the desired
material.
(c) N-Ms-Phe-Gly-Ala-LeuH: A solution is prepared by adding 0.337 g (0.798
mmol) Dess-Martin periodinane to S ml CH2C12 and stirring for 10 minutes. To this
solution 0.125 g (0.266 mmol) N-Ms-Phe-Gly-Ala-Leucinol is added in one portion.The reaction is continued until TLC showed complete conversion at which time thesolution is poured into 25 ml sat'd NaHCO3 cont~inin~ 1.8 g (5.586 mmol)
Na2S2O3. After stirring for 10 minutes the mixture is extracted with EtOAc (3X, 50
ml). The combined extracts are dried (MgSO4), evaporated, and chromatographed
on silica to afford 0.048 g of the product.
Synthesis Example 5:
SYnthesis of an aldeh~lde protease inhibitor
Moc-Leu-OH-L-Leucine (5.0 g, 38.2 mmol) is dissolved in 38 ml lN NaOH and
cooled to 0~C. Methyl chloroformate (3.1 ml, 40.0 mmol) is added dropwise while
in a separate addition funnel lN NaOH is added as to m~int~in pH at 9.0-9.5. After
addition is complete and the pH stabilized at 9.0-9.5 the solution is washed with 200
ml EtOAc, the aqueous phase is then acidified to pH = 2. This mixture is extracted
with EtOAc (2X 100 ml), dried (MgSO4), filtered, and the solvent removed to afford
7.15 g pure product.
Moc-Leu-Leucinol- To a solution of 3.5 g (18.52 mmol) Moc-Leu-OH in 100 ml
THF, cooled to -15~C, 2.04 ml (18.52 mmol) of N-methyl morpholine is added
followed imme~ te~ly by 2.4 ml (18.52 mmol) isobutyl chloroformate. After
stirring for 10 minutes 2.37 ml (18.52 mmol) of leucinol in 25 ml of THF is added
and the reaction stirred 0.5 h at -15~C and 1 h at room tc~ ,e.~l lre. The mixture is
then diluted with 100 ml of H20 and the THF evaporated. The re.n~ining a~ueous
phase is partitioned between EtOAc and lN HCl, the organic phase washed with
NaHCO3, dried (MgSO4) and evaporated to afford 5.33 g pure product.
Moc-Leu-LeuH-A solution cont~ining 4.4 g (10.41 mmol) Dess-Martin periodinane
suspended in 100 ml CH2C12 is pre~ ed and stirred for 10 minutes. To this solution
1.0 g ~3.47 mmol) Moc-Leu-Leucinol is added and the solution stirred 2 h at roomt~ pc.dlllre followed by pouring into 100 ml of saturated NaHCO3 cont~ining 18 g(72.87 mmol) Na2S2O3. This solution is stirred 10 minnt~c and then extracted with
CA 02266~27 1999-03-23
WO 98/13459 PCTIUS97/16622
13
..
EtOAc (2X, 125ml), dried (MgSO4) and the solvent evaporated. Chromatography
on silica affords 0.550 g of pure product.
Synthesis Exarnple 6:
Additional peptide aldehydes are synth~i7e~1 according to the following
procedures. Some of the interme.li~tçc are purchased from suppliers and in theseinct~nf~es it is noted within the procedure. Dess-Martin periodinane is synthesized
according to the procedure of Martin, J.Org. Chem., 1983, 48, 4155.
1 O I. Z-Gly-Ala-Leu-OMe - To a solution of Z-Gly-Ala-OH (20.0 g, 0.071 M) and
Leu-OMe.HCl (12.9 g, 0.071 M) in 250 ml dichloromethane is added 21.9 ml (0.157
M) triethylamine (TEA) dropwise over a period of 10 min. This addition is followed
by the addition of 11.9 ml (0.078 M) of diethylcyanophosphonate (DECP). The
mixture is stirred overnight and the solvent removed. The residue is dissolved in
ethyl acetate and washed with lN HCl, saturated NaHCO3, and brine. The solution
is dried with MgSO4, filtered and the solvent removed. Recovered will be 29.0 g of
product that is homogeneous by TLC. 13C NMR (CDC13) 15.93, 18.60, 21.77,
22.69, 24.72, 40.80, 44.20, 48.70, 50.87, 52.13, 65.28, 66.84, 127.92, 128.00,
128.41, 136.36, 156.76, 169.31, 172.58, 173.24.
II. Moc-Phe-Gly-Ala-Leu-OMe - Z-Gly-Ala-Leu-OMe (29.0 g, 0.071 M) is
dissolved in 300 ml MeOH and 35 ml 4.0 M HCI in dioxane. To this solvent
mixture is added 5.8 g of 10% Pd/C portionwise. The slurry is ~eg~csed with an
aspirator and H2 introduced via balloon. The slurry is m~int~in~d under a positive
pressure of H2 and stirred overnight. The slurry is filtered through Celite and a
sintered glass funnel and washed thoroughly with MeOH. The solvent is removed
and the residue is triturated with ether. The slurry is filtered and the filter cake dried
under vacuurn. Recovered 20.2 g of an off-white powder. The crude product and
Moc-Phe-OH (15.3 g, 0.068 M) are dissolved in 500 ml CH2Cl2 and 29.9 ml TEA
(0.143 M) added dropwise followed by the dropwise addition of 11.7 ml (0.072 M)
of DECP. The mixture iss stirred overnight and the solvent is removed. The residue
is dissolved in EtOAc and washed with lN HCI, saturated NaHCO3, and brine. The
organic phase is dried (MgS04), filtered and the solvent removed to afford 21.3 g
product. 13C NMR (CDC13) 16.66, 16.83, 20,01, 22.46, 23.41, 25.40, 40.11, 41.72,43.75, 49.39, 51.37, 52.87, 56.42, 65.92, 77.39, 77.55, 77.81, 78.24, 127.42, 128.96,
129.19, 130.09, 137.41, 157.62, 169.00, 172.63, 173.24, 174.00.
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WO 98/134S9 PCT/US97/16622
14
III. Moc-Phe-Gly-Ala-Leucinol - Moc-Phe-Gly-Ala-Leu-OMe (21.3 g, 44.5 mmol)
is dissolved in a mixture of 400 ml EtOH and 250 ml THF. The solution is cooled
to 0~C and 9.88 g (89.0 mmol) CaC12 is added. In 5 min the slurry will be
homogenized and 6.73 g (178.0 mmol) NaBH4 added portionwise over a period of 5
5 min. The solution is stirred at 0~C for 2 hours and the reaction carefully quenched
with lN HCI. The EtOH and THF are removed under vacuum and the rem~ining
aqueous mixture extracted with 500 ml EtOAc. This organic phase is washed with
saturated NaHCO3, brine, and the organic phase dried with MgSO4. Filtration and
removal of solvent affords 20.0 g of an off-white crystalline material.
l o Chromatography on silica (3.5% MeOH/CH2C12) gives 13.0 g pure product. Rf =
0.3 (10% MeOH/CH2Cl2), 13C NMR (CDC13) 17.50, 22.23, 23.12, 24.84, 37.22,
39.76, 43.96, 49.88, 50.93, 52.48, 58.22, 65.27, 98.46, 98.54, 127.04, 128.68,
129.10, }36.62, 157.85, 170.71, 173.85, 174.45
1 5 IV. Moc-Phe-Gly-Ala-Leu-H - 29.9 g (70.7 mmol) of Dess-Martin periodinane is
suspended in 500 ml CH2C12 and stirred for 10 min. Moc-Phe-Gly-Ala-Leucinol
(10.6 g, 23.5 mmol) iss dissolved in 100 ml C1~2C12 and added at a moderate rate to
the periodinane slurry. The mixture is stirred for Ih and poured into 150 ml
NaHCO3 cont~ining 123 g Na2S2O3. The mixture is allowed to stir for 15 min and
20 extracted with EtOAc. The organic phase is dried and filtered followed by removal
of solvent. Chromatography (3.5% MeOH/CH2C12) on silica gives 5.1 g of pure
white solid that is a mixture of the methoxy hemi~ret~l and aldehyde. 13C NMR
(CDC13,CD30D) 17.62, 17.94, 21.53, 21.71, 22.99, 23.30, 23.39, 24.54, 37.05,
37.70, 37.92, 38.24, 42.87, 49.83, 51.79, 52.14, 52.40, 56.75, 57.19, 98.40, 99.18,
25 127.00, 128.60, 129.06, 136.44, 157.27, 169.19, 169.67, 172.73, 173.40, 200.43.
V. Moc-Phe-OH - E-Phenylalanine (5.0 g, 30.2 rnmol) is dissolved in 30 ml lN
NaOH and cooled to 0~C. Methyl chloroformate (2.53 ml, 31.8 mmol) is added
dropwise while in a sep~ate addition funnel 30 ml of lN NaOH is added
30 simultaneously. After addition is complete, the solution is washed with 200 ml
EtOAc and the aqueous phase acidified to pH = 2. The mixture is extracted with
EtOAc (2X 100 ml), dried (MgSO4), filtered, and the solvent removed to afford 6.0
g product. 13C NMR (CDC13) 37.75, 52.57, 54.64, 128.63, 129.35, 135.74, 156.77,
175.76.
VI. Mac-Phe-OH - To a solution of 1.00 g (2.34 mmol) of Phe-OBn.PTSA in Et2O
at room ten,p~.dl~Ire is added 0.36 ml (2.57 mmol) of TEA. This is followed by the
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WO 98/13459 ~CT/US97/16622
addition of 10 ml MeOH and then 0.14 ml (2.34 mmol) of methyl isocyanate in 4 mlEt2O is added dropwise. The reaction mixture is poured into 50 ml water and the
phases separate. The organic phase is dried with MgSO4, filtered and the solventremoved to give 0.66 g of product (96% yield). 13C NMR (CDC13) 27.05, 38.47,
53.45, 54.64, 65.90, 127.43, 127.85, 128.48, 129.28, 130.21, 135.23, 136.22, 158.17,
173.08. To a solution of the crude product (2.11 mmol) in 25 ml MeOH is added
0.120 g Pd/C and the slurry deg~cse(l The slurry is stirred under a positive pressure
of H2 via balloon for 1.5 h. The slurry is filtered through Celite and the filter cake
washed with MeOH. The solvent is removed to afford 0.430 g product. 13C NMR
0 26.50, 37.92, 54.28, 126.69, 128.28, 129.28, 136.65, 159.36, 175.33.
VII. Mac-Phe-Gly-Ala-Leucinol - To a solution of 0.200 g Mac-Phe-OH (0.900
mmol) and 0.253 g Gly-Ala-Leu-OMe.HCI (0.818 mmol, generated by
hydrogenation of I., above, according to the procedure outlined for compound II.) in
15 ml DMF is added 0.250 ml TEA (1.80 mmol) followed by the addition of 0.147
ml DECP (0.900 mmol). The mixture is stirred overnight and the solvent removed.
The residue is redissolved in EtOAc and washed successively with 0.3 N HCI,
saturated NaHCO3, and brine. The solution is dried, filtered and the solvent
removed to give 0.300 g product. The crude product (0.628 mmol) is dissolved in
17 ml EtOH and cooled to 0~C. To this solution is added 0.140 g CaCl2 (1.25
mmol) in 4 ml THF. To the resulting slurry is added 0.095 g NaBH4 in one portion.
After 45 min. the solution is quenched with water and extracted with EtOAc. The
organic phase is dried with MgSO4, filtered and the solvent removed.
Chromatography with 4% MeOH/CH2C12 gave 0.200 g pure product. 13C NMR
(CD30D) 16.84, 21.05, 22.60, 24.51, 25.66, 37.41, 39.73, 42.67, 49.65, 56.63,
64.33, 126.63, 128.32, 128.96, 137.12, 160.01, 170.45, 173.60, 175.03.
VIII. Mac-Phe-Gly-Ala-Leu-H - To a slurry of Dess-Martin periodinane (0.565 g,
1.33 mmol) in 15 ml CH2C12 is added a suspension of Mac-Phe-Gly-Ala-Leucinol
(0.200 g, 0.445 mmol) in CH2Cl2 and the resulting slurry stirred for 0.5 h. The
mixture is poured into saturated NaHC03 col-t~ i..g 2.32 g Na2S203 and the
- solution stirred for 10 min., followed by extraction ~vith EtOAc. The organic phase
is dried with MgS04, filtered and the solvent removed. The residue is
chromatographed on silica to give 0.081 g product. 13C NMR (10% CD30D in
CDC13) 17.18, 17.43, 21.35, 21.55, 23.26, 23.34, 24.40, 24.47, 26.36, 26.60, 37.25,
37.38, 38.60, 42.86, 42.97, 51.77, 51.93, 54.94, 56.75, 57.00, 98.7, 99.32, 126.87,
128.49, 128.91, 136.51, 159.53, 159.55, 169.93, 170.39, 173.63, 173.85, 174.70.
CA 02266~27 1999-03-23
WO 98/134S9 PCT/US97/16622
16
Cbz= carbobenzyloxy
Gly = glycine
Ala= alanine
5 Leu = leucine
Phe = phenylalanine
OMe = methyl ester
TEA = triethylamine
DECP = diethylcyanophosphonate
10 TLC = thin layer chromatography
MeOH = methanol
Pd/C = palladium on activated carbon
EtOH = ethanol
THF = tetrahydrofuran
Mac = methylaminocarbonyl
Moc = methoxycarbonyl
Etoc = ethoxycarbonyl
Ms = meth~nesulfonyl
Proteolytic ~nz,vme - Another essential ingredient in the present liquid
detergent compositions is active proteolytic enzyme. Mixtures of proteolytic
enzyme are also included. The proteolytic enzyme can be of animal, vegetable or
microorganism (preferred) origin. The proteases for use in the detergent
compositions herein include (but are not limited to) trypsin, subtilisin, chymotrypsin
and elastase-type proteases. Preferred for use herein are subtilisin-type proteolytic
enzymes. Particularly p~cr~ . d is bacterial serine proteolytic enzyme obtained from
Bacillus subtilis and/or Bacillus licheniforrnis. Protease enzymes are usually present
in such liquid detergent compositions at levels sufficient to provide from 0.005 to
0.1 Anson units (AU) of activity per grarn of composition.
Suitable proteolytic enzymes include Novo Industri A/S Alcalase~) (preferred),
Esperase~), Savinase~ (Copenhagen, Del~n~k), Gist-brocades' Maxatase~,
Maxacal~ and Maxapem 1 5~g~ (protein engineered Maxacal(~)) (Delft, Netherlands),
and subtilisin BPN and BPN'(prcfe.lcd), which are commercially available.
Preferred proteolytic enzymes are also modified bacterial serine proteases, such as
3~ those made by Genencor Int~rn~tional, Inc.(San Francisco, California) which are
described in European Patent 251,446, filed April 28, 1987 (particularly pages 17,
24 and 98), and which is called herein "Protease B", and U.S. Patent 5,030,378,
CA 02266~27 1999-03-23
W O 98/134S9 PCTAUS97/16622
17
Venegas, issued July 9, 1991, which refers to a modified bacterial serine proteolytic
enzyme (Genencor International) which is called "Protease A" herein (same as
BPN'). In particular see columns 2 and 3 of U.S. Patent 5,030,378 for a completedescription, including amino sequence, of Protease A and its variants. Preferredproteolytic enzymes, then, are selected from the group consisting of Alcalase (~)
(Novo Industri A/S), BPN', Protease A and Protease B (Genencor), and mixtures
thereof. Protease B is most preferred.
Another preferred protease, referred to as "Protease D" is a carbonyl
hydrolase variant having an amino acid sequence not found in nature, which is
10 derived from a precursor carbonyl hydrolase by substituting a dirr~.elll amino acid
for a plurality of amino acid residues at a position in said carbonyl hydrolase
equivalent to position +76, preferably also in combination with one or more amino
acid residue positions equivalent to those selected from the group con~ictine of +99,
+101, +103, +104, +107, +123, +27, +105, +109, +126, +128, +135, +156, +166,
15 +195, +197, +204, +206, +210, +216, +217, +21~, +222, +260, +265, and/or +274according to the numbering of Bacillus amyloliquefaciens subtilisin, as described in
WO 95/10615 published April 20, 1995 by Genencor International.
Useful proteases are also described in PCT publications: WO 95/30010
published Novenber 9, 1995 by The Procter & Gamble Company; WO 95/30011
20 published Novenber 9, 1995 by The Procter & Gamble Company; WO 95/29979
published Novenber 9, 1995 by The Procter & Gamble Company.
Calcium - Any water-soluble calcium salt can be used as a source of calcium
ions, including calcium acetate, calcium formate, calcium xylene sulfonate, and
calcium propionate. Divalent ions, such as zinc and magnesium ions, can replace
25 the calcium ion completely or in part. Thus in the liquid detergent compositions
herein, the source of calcium ions can be partially substituted with a source ofanother divalent ion.
The calcium useful herein is enzyme-~ccessihle. Therefore, the cl~imçd
compositions are substantially free of sequestrants, for example, polyacids capable
30 of forming calcium complexes which are soluble in the composition. However,
minor amounts of sequestrants such as polyacids or mixtures of polyacids can be
used. The enzyme-accessible calcium is defined as the amount of calcium-ions
effectively available to the enzyme component. From a practical standpoint the
enzyme-accessible calcium is therefore the soluble calcium in the composition in the
35 absence of any storage sequestrants, e.g., having an equilibrium constant of
complexation with calcium equal to or greater than 1.5 at 20~C.
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W O 98/13459 PCTrUS97/16622
18
Boric Acid - The compositions herein optionally contain from about 0.25% to
about 10%, preferably from about 0.5% to about 5%, more preferably from about
0.75% to about 3%, by weight of boric acid or a compound capable of forming boric
acid in the composition (calculated on the basis of the boric acid). Boric acid is
preferred, although other compounds such as boric oxide, borax and other alkali
metal borates (e.g., sodium ortho-, meta-, pyroborate, an sodium pentaborate) are
suitable. Substituted boric acids (e.g., phenylboronic acid, butane boronic acid, and
p-bromo phenylboronic acid) can also be used in place of boric acid.
The compositions of the present invention can also contain polyols, especially
10 diols, cont~ining only carbon, hydrogen and oxygen atoms. They preferably contain
from about 2 to about 6 hydroxy groups. Examples include propylene glycol
(especially 1,2 propanediol, which is preferred), ethylene glycol, glycerol, sorbitol,
mannitol, glucose, and mixtures thereof. The polyol generally represents from about
1% to about 15%, preferably from about 1.5% to about 10%, more preferably from
about 2% to about 7%, by weight of the composition.
Detersive Surfactant - An effective amount, typically from about I to 95,
preferably about 8 to 70, weight %, of detersive surfactant is yet another escenti~l
ingredient in the present invention. The detersive surfactant can be selected from the
group concicting of anionics, nonionics, cationics, ampholytics, zwitterionics, and
20 mixtures thereof. By selecting the type and amount of detersive surfactant, along
with other adjunct ingredients disclosed herein, the present detergent compositions
can be formulated to be used in the context of laundry cleaning or in other different
cleaning applications, particularly including dishwashing. The particular surfactants
used can therefore vary widely depending upon the particular end-use envisioned.The benefits of the present invention are especially pronounced in
compositions cont~ining ingredients that are harsh to enzymes such as certain
detergency builders and surf~rt~ntc These include (but are not limited to) anionic
surfactants such as alkyl ether sulfate linear alkyl benzene sulfonate, alkyl sulfate,
etc. Suitable surfactants are described below.
Anionic Surfactants - One type of anionic surfactant which can be utilized
encomp~ccec alkyl ester sulfonates. These are desirable because they can be madewith renewable, non-petroleum resources. Preparation of the alkyl ester sulfonate
surfactant component can be effected according to known methods disclosed in thetechnical literature. For inct~n~e, linear esters of Cg-C20 carboxylic acids can be
35 sulfonated with gaseous S03 according to "The Journal of the American Oil
Chemists Society," 52 (1975), pp. 323-329. Suitable starting materials would
include natural fatty substances as derived from tallow, palm, and coconut oils, etc.
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W O 98/134S9 PCTrUS97/16622
19
The preferred alkyl ester sulfonate surfactant, especially for laundry
applications, comprises alkyl ester sulfonate surfactants of the structural forrnula:
o
R3 -CH-C-oR4
S03M
wherein R3 is a Cg-C20 hydrocarbyl, preferably an alkyl, or combination thereof,R4 is a C 1 -C6 hydrocarbyl, preferably an alkyl, or combination thereof, and M is a
soluble salt-forming cation. Suitable salts include metal salts such as sodium,
potassium, and lithium salts, and substituted or unsubstituted ammoniurn salts, such
0 as methyl-, dimethyl, -trimethyl, and quaternary ammonium cations, e.g.
tetramethyl-ammonium and dimethyl piperdinium, and cations derived from
alkanolAmines, e.g. monoethanol-amine, diethanolamine, and triethanolamine.
Preferably, R3 is Clo-Cl6 alkyl, and R4 is methyl, ethyl or isopropyl. Especially
preferred are the methyl ester sulfonates wherein R3 is C 1 4-C 16 alkyl.
Alkyl sulfate surfactants are another type of anionic surfactant of importance
for use herein. In addition to providing excellent overall cleaning ability when used
in combination with polyhydroxy fatty acid amides (see below), including good
grease/oil cleaning over a wide range of tel,lpeldlures, wash concentrations~ and
wash times, dissolution of alkyl sulfates can be obtained, as well as improved
formulability in liquid detergent formulations are water soluble salts or acids of the
formula ROSO3M wherein R preferably is a C l o-C24 hydrocarbyl, preferably an
alkyl or hydroxyalkyl having a C l o-c20 alkyl component, more preferably a C 12-
C 1 8 alkyl or hydroxyalkyl, and M is H or a cation, e.g., an alkali metal cation (e.g.,
sodium, potassium, lithium), sub~liluled or unsubstituted ammonium cations such as
methyl-, dimethyl-, and trimethyl Ammonium and qUAtenlAry ammonium cations,
e.g., t~llalll~;l}lyl-ammonium and dimethyl piperdinium, and cations derived from
alkanolAmines such as ethanolamine, diethanolamine, triethanolamine, and mixtures
thereof, and the like. Typically, alkyl chains of C 12-16 are preferred for lower wash
t~ pcldlules (e.g., below about 50~C) and C16 18 alkyl chains are p~ ed for
higher wash telll~ alules (e.g., above about 50~C).
Alkyl alkoxylated sulfate surfAct~nt~ are another category of useful anionic
surfactant. These surfAct~nt~ are water soluble salts or acids typically of the formula
RO(A)mSO3M wherein R is an unsubstituted Clo-C24 alkyl or hydroxyalkyl group
having a C1o-C24 alkyl component, preferably a C12-C20 alkyl or hydroxyalkyl,
more preferably C12-C18 alkyl or hydroxyalkyl, A is an ethoxy or propoxy unit, mis greater than zero, typically between about 0.5 and about 6, more preferably
between about 0.5 and about 3, and M is H or a cation which can be, for example, a
CA 02266~27 1999-03-23
WO 98/13459 PCT/US97/16622
metal cation (e.g., sodium, potassium, lithium, calcium, magnesium, etc.),
ammoniurn or substituted-ammonium cation. Alkyl ethoxylated sulfates as well as
- alkyl propoxylated sulfates are contemplated herein. Specific exarnples of
substituted ammonium cations include methyl-, dimethyl-, trimethyl-amrnonium and5 quaternary ammonium cations, such as tetramethyl-ammonium, dimethyl
piperidinium and cations derived from alkanol~min.-s, e.g. monoethanolamine,
diethanolamine, and triethanolamine, and mixtures thereof. Exemplary surfactantsare C12-CIg alkyl polyethoxylate (1.0) sulfate, C12-Clg alkyl polyethoxylate (2.25)
sulfate, C 12-c 18 alkyl polyethoxylate (3.0) sulfate, and C 12-c 18 alkyl
1 o polyethoxylate (4.0) sulfate wherein M is conveniently selected from sodium and
potassium.
Other Anionic Surfactants - Other anionic surfactants useful for detersive
purposes can also be included in the compositions hereof. These can include salts
(including, for example, sodium, potassium, ammonium, and substituted amrnonium
15 salts such as mono-, di- and triethanolamine salts) of soap, Cg-C20 linear
alkylben7erlesnll honates, Cg-C22 primary or secondary alkanesulphonates, Cg-C24olefinsulphonates, sulphonated polycarboxylic acids prepared by sulphonation of the
pyrolyzed product of ~Ik~lin~ earth metal citrates, e.g., as described in British patent
specification No. 1,082,179, alkyl glycerol sulfonates, fatty acyl glycerol sulfonates,
20 fatty oleyl glycerol sulfates, alkyl phenol ethylene oxide ether sulfates, paraffin
sulfonates, alkyl phosphates, isothionates such as the acyl isothionates, N-acyltaurates, fatty acid amides of methyl tauride, alkyl succin~m~tes and sulfosuccinates,
monoesters of sulfosuccinate (especially saturated and unsaturated C 12-c 18
monoesters) diesters of sulfosuccinate (especially saturated and unsaturated C6-C 14
25 diesters), N-acyl sarcosin~es, sulfates of alkylpolysaccharides such as the sulfates of
alkylpolyglucoside (the nonionic non~lllf~te~l compounds being described below),blanched primary alkyl snlf~t~s, alkyl polyethoxy carboxylates such as those of the
formula ~O(CH2CH20)kCH2COO-M+ wherein R is a Cg-C22 alkyl, k is an
integer from O to 10, and M is a soluble salt-forming cation, and fatty acids
30 esterified with isethionic acid and neutralized with sodium hydroxide. Resin acids
and hydrogen~te~ resin acids are also suitable, such as rosin, hydrogenated rosin,
and resin acids and hydrogenated resin acids present in or derived from tall oil.
Further exarnples are given in "Surface Active Agents and Detergents" (Vol. I and II
by Schwartz, Perry and Berch). A variety of such surfactants are also generally
3~ disclosed in U.S. Patent 3,929,678, issued December 30, 1975 to ~,~ughlin, et al. at
Column 23, line 58 through Column 29, line 23 (herein incorporated by reference).
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21
Nonionic Deter~ent Surfactants - Suitable nonionic detergent surfactants are
generally disclosed in U.S. Patent 3,929,678, I.~llghlin et al., issued December 30,
1975, at column 13, line 14 through column 16, line 6, incorporated herein by
reference. Exemplary, non-limiting classes of useful nonionic surfactants are listed
below
The polyethylene, polypropylene, and polybutylene oxide condensates of alkyl
phenols. In general, the polyethylene oxide con~lenc~tes are preferred. These
compounds include the con-lenc~tion products of alkyl phenols having an alkyl
group cont~ining from about 6 to about 12 carbon atoms in either a straight chain or
branched chain configuration with the alkylene oxide. In a preferred embodiment,the ethylene oxide is present in an amount equal to from about 5 to about 25 moles
of ethylene oxide per mole of alkyl phenol. Commercially available nonionic
surfactants of this type include Igepal~ C0-630, marketed by the GAF Corporation;
and Triton~) X-45, X-l 14, X-100, and X-102, all marketed by the Rohm & Haas
Company. These compounds are commonly referred to as alkyl phenol alkoxylates,
(e.g., alkyl phenol ethoxylates).
The conrienc~tion products of aliphatic alcohols with from about 1 to about 25
moles of ethylene oxide. The alkyl chain of the aliphatic alcohol can either be
straight or branched, primary or secondary, and generally contains from about 8 to
about 22 carbon atoms. Particularly p~er~,~cd are the condensation products of
alcohols having an alkyl group cont~ining from about 10 to about 20 carbon atomswith from about 2 to about 18 moles of ethylene oxide per mole of alcohol.
Examples of co,.,.,lelcially available nonionic surf~ct~ntc of this type includeTergitol~) 1 5-S-9 (the conden.c~tion product of C 1 1 -C 15 linear secondary alcohol
with 9 moles ethylene oxide), Tergitol(g) 24-L-6 NMW (the con~en.c~tion product of
C12-C14 primary alcohol with 6 moles ethylene oxide with a narrow molecular
weight distribution), both marketed by Union Carbide Corporation; Neodol(~ 45-9
(the con~len~tion product of C 1 4-C 15 linear alcohol with 9 moles of ethylene
oxide), Neodol~ 23-6.5 (the con~ien~tion product of C 1 2-C 13 linear alcohol with
6.5 moles of ethylene oxide), Neodol(~ 45-7 (the con~en~tion product of C 1 4-C 15
linear alcohol with 7 moles of ethylene oxide), Neodol~) 45-4 (the con-len.c~tion
product of C 1 4-C 15 linear alcohol with 4 moles of ethylene oxide), marketed by
Shell Chemical Col"pal1y, and Kyro~) EOB (the condPnc~tion product of C 1 3-C 15alcohol with 9 moles ethylene oxide), marketed by The Procter & Gamble Company.
This category of nonionic surfactant is referred to generally as "alkyl ethoxylates."
The con-len~tion products of ethylene oxide with a hydrophobic base fonned
by the conden~tion of propylene oxide with propylene glycol. The hydrophobic
, ... _ ... .. . . ....
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22
portion of these compounds preferably has a molecular weight of from about 1500 to
about 1800 and exhibits water insolubility. The addition of polyoxyethylene
moieties to this hydrophobic portion tends to increase the water solubility of the
molecule as a whole, and the liquid character of the product is retained up to the
point where the polyoxyethylene content is about 50% of the total weight of the
con~c~tion product, which corresponds to con.1enc~tion with up to about 40 molesof ethylene oxide. Examples of compounds of this type include certain of the
commercially-available Pluronic(~) surf~.~t~nt.~, marketed by BASF.
The con-len~tion products of ethylene oxide with the product resulting from
0 the reaction of propylene oxide and ethylene.1i~mine. The hydrophobic moiety of
these products consists of the reaction product of ethylene~ rnin~- and excess
propylene oxide, and generally has a molecular weight of from about 2500 to about
3000. This hydrophobic moiety is condensed with ethylene oxide to the extent that
the condensation product contains from about 40% to about 80% by weight of
polyoxyethylene and has a molecular weight of from about 5,000 to about 11,000.
Examples of this type of nonionic surfactant include certain of the cornmercially
available Tetronic~ compounds, marketed by BASF.
Semi-polar nonionic surf~ct~nt.c are a special category of nonionic surfactants
which include water-soluble amine oxides cont~ining one alkyl moiety of from
20 about 10 to about 18 carbon atoms and 2 moieties selected from the group consisting
of alkyl groups and hydroxyalkyl groups cont~ining from about 1 to about 3 carbon
atoms; water-soluble phosphine oxides cont~ining one alkyl moiety of from about 10
to about 18 carbon atoms and 2 moieties selected from the group consisting of alkyl
groups and hydroxyalkyl groups cont~inin~ from about I to about 3 carbon atoms;
25 and water-soluble sulfoxides CO~ ing one alkyl moiety of from about 10 to about
18 carbon atoms and a moiety selected from the group consisting of alkyl and
hydroxyalkyl moieties of from about 1 to about 3 carbon atoms.
Semi-polar nonionic detergent surfactants include the arnine oxide surfactants
having the forrnula
O
R3(oR4)XN(R5)2
wherein R3 is an alkyl, hydroxyalkyl, or alkyl phenyl group or mixtures thereof
cont~inin~ from about 8 to about 22 carbon atoms; R4 is an alkylene or
hydroxyalkylene group cont~ining from about 2 to about 3 carbon atoms or mixtures
35 thereof; x is from 0 to about 3; and each R5 is an alkyl or hydroxyalkyl group
cont~ining from about I to about 3 carbon atoms or a polyethylene oxide group
cont~ining from about 1 to about 3 ethylene oxide groups. The R5 groups can be
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23
rhP~l to each other, e.g., through an oxygen or nitrogen atom, to form a ring
structure.
These amine oxide surfactants in particular include C 1 o-C 18 alkyl dimethyl
amine oxides and Cg-C12 alkoxy ethyl dihydroxy ethyl amine oxides.
Alkylpolysaccharides disclosed in U.S. Patent 4,565,647, Llenado, issued
January 21, 1986, having a hydrophobic group cont~ining from about 6 to about 30carbon atoms, preferably from about 10 to about 16 carbon atoms and a
polysaccharide, e.g., a polyglycoside, hydrophilic group cont~ining from about 1.3
to about 10, preferably from about 1.3 to about 3, most preferably from about 1.3 to
about 2.7 saccharide units. Any reducing saccharide cont~ining 5 or 6 carbon atoms
can be used, e.g., glucose, galactose and galactosyl moieties can be substituted for
the glucosyl moieties. (Optionally the hydrophobic group is attached at the 2-, 3-, 4-,
etc. positions thus giving a glucose or galactose as opposed to a glucoside or
galactoside.) The intersaccharide bonds can be, e.g., between the one position of the
additional saccharide units and the 2-, 3-, 4-, and/or 6- positions on the preceding
saccharide units.
Optionally, and less desirably, there can be a polyalkylene-oxide chain joining
the hydrophobic moiety and the polysaccharide moiety. The preferred alkyleneoxide
is ethylene oxide. Typical hydrophobic groups include alkyl groups, either saturated
or unsaturated, branched or unbranched cont~ining from about 8 to about 18,
preferably from about 10 to about 16, carbon atoms. Preferably, the alkyl group is a
straight chain saturated alkyl group. The alkyl group can contain up to about 3
hydroxy groups and/or the polyalkyleneoxide chain can contain up to about 10,
preferably less than 5, alkyleneoxide moieties. Suitable alkyl polysaccharides are
octyl, nonyl, decyl, undecyldodecyl, tridecyl, tetradecyl, pent~r~Pcyl, hexadecyl,
heptadecyl, and octadecyl, di-, tri-, tetra-, penta-, and hexaglucosides, galactosides,
lactosides, glucoses, fructosides, fructoses and/or galactoses. Suitable mixtures
include coconut alkyl, di-, tri-, tetra-, and pentaglucosides and tallow alkyl tetra-,
penta-, and hexa-glucosides.
The pr~felled alkylpolyglycosides have the formula
R20(CnH2nO)t(glYC~sYl)x
wherein R2 is selected from the group consisting of alkyl, alkyl-phenyl,
hydroxyalkyl, hydroxyalkylphenyl, and mixtures thereof in which the alkyl groupscontain from about 10 to about 18, preferably from about 12 to about 14, carbon
atoms; n is 2 or 3, preferably 2; t is from 0 to about 10, preferably 0; and x is from
about 1.3 to about 10, preferably from about 1.3 to about 3, most preferably from
about 1.3 to about 2.7. The glycosyl is preferably derived from glucose. To prepare
, ~ . . . . . . . ...
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24
these compounds, the alcohol or alkylpolyethoxy alcohol is forrned first and then
reacted with glucose, or a source of glucose, to form the glucoside (attachment at the
l-position). The additional glycosyl units can then be attached between their 1-position and the prece~ling glycosyl units 2-, 3-, 4- and/or 6-position, preferably
5 predominantly the 2-position.
Fatty acid amide surf~ct~nt~ having the formula:
o
R6 -C-N (R7 ) 2
wherein R6 is an alkyl group cont~inine from about 7 to about 21 (preferably from
10 about 9 to about 17) carbon atoms and each R7 is selected from the group consisting
of hydrogen, C 1 -C4 alkyl, C I -C4 hydroxyalkyl, and -(C2H40)XH where x varies
from about 1 to about 3.
Preferred amides are Cg-C20 ammonia amides, monoethanolamides,
diethanolamides, and isopropanol~mi~les
Cationic Surf~ct~nt.~ - Cationic detersive surfactants can also be included in
detergent compositions of the present invention. Cationic surfactants include the
ammonium surfactants such as alkyldimethylammonium halogenides, and those
surfactants having the formula:
[R2(oR3 )y] [R4(oR3 )y]2R5N+X-
wherein R2 is an alkyl or alkyl benzyl group having from about 8 to about 18 carbon
atoms in the alkyl chain, each R3 is selected from the group consisting of-CH2CH2-
-CH2CH(CH3)-, -CH2CH(CH2OH)-, -CH2CH2CH2-, and mixtures thereof; each
R4 is selected from the group con~i~ting of Cl-C4 alkyl, Cl-C4 hydroxyalkyl,
benzyl, ring structures formed by joining the two R4 groups, -
CH2CHOHCHOHCOR6CHOH-CH20H wherein R6 is any hexose or hexose
polymer having a molecular weight less than about 1000, and hydrogen when y is
not O; R5 is the same as R4 or is an alkyl chain wherein the total number of carbon
atoms of R2 plus RS is not more than about 18; each y is from 0 to about 10 and the
sum of the y values is from 0 to about 15; and X is any compatible anion.
Other cationic surfA~-tAnt~ useful herein are also described in U.S. Patent
4,228,044, Cambre, issued October 14, 1980, incorporated herein by reference.
Other Surfactants - Ampholytic surf~ct~nt~ can be incorporated into the
detergent compositions hereof. These surfactants can be broadly described as
aliphatic derivatives of secondary or tertiary amines, or aliphatic derivatives of
heterocyclic secondary and tertiary amines in which the aliphatic radical can bestraight chain or branched. One of the aliphatic substituents contains at least about 8
carbon atoms, typically from about 8 to about 18 carbon atoms, and at least one
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contains an anionic water-solubilizing group, e.g., carboxy, sulfonate, sulfate. See
U.S. Patent No. 3,929,678 to L ~ghlin et al., issued December 30, 1975 at column19, lines 18-35 forexamples of ampholytic surfactants.
Zwitterionic surfactants can also be incorporated into the detergent
compositions hereof. These surfactants can be broadly described as derivatives of
secondary and tertiary amines, derivatives of heterocyc}ic secondary and tertiary
amines, or derivatives of quat~ arnmonium, quaternary phosphonium or tertiary
sulfonium compounds. See U.S. Patent No. 3,929,678 to T ~llghlin et al., issued
December 30, 1975 at column 19, line 38 through column 22, line 48 for examples
10 of zwitterionic surf~ct~ntc Arnpholytic and zwitterionic surf~ct~nts are generally
used in combination with one or more anionic and/or nonionic surfactants.
PolYhydroxy Fatty Acid Amide Surfactant - The liquid detergent compositions
hereof may also contain an enzyme-enhancing amount of polyhydroxy fatty acid
amide surfactant. By "enzyme-enhancing" is meant that the formulator of the
15 composition can select an amount of polyhydroxy fatty acid amide to be
incorporated into the compositions that will improve enzyme cleaning performanceof the detergent composition. In general, for conventional levels of enzyme, theincorporation of about 1 %, by weight, polyhydroxy fatty acid amide will enhanceenzyme performance.
The detergent compositions herein will typically comprise about 1% weight
basis, polyhydroxy fatty acid amide surfactant, preferably from about 3% to about
30%, of the polyhydroxy fatty acid arnide. The polyhydroxy fatty acid amide
surfactant component comprises compounds of the structural formula:
O Rl
R2 - C - N - z
wherein: Rl is H, Cl-C4 hyd~oc~l,yl, 2-hydroxy ethyl, 2-hydroxy propyl, or a
mixture thereof, p~fcl~bly C1-C4 alkyl, more preferably C1 or C2 alkyl, most
preferably C I alkyl (i.e., methyl); and R2 is a Cs-C31 hydrocarbyl, preferably
straight chain C7-C 19 alkyl or alkenyl, more preferably straight chain Cg-C 17 alkyl
30 or alkenyl, most preferably straight chain C 11 -C 15 alkyl or alkenyl, or mixtures
thereof; and Z is a polyhydroxyhydrocarbyl having a linear hydrocarbyl chain with
at least 3 hydroxyls directly co~nectecl to the chain, or an alkoxylated derivative
(preferably ethoxylated or propoxylated) thereof. Z preferably will be derived from
a reducing sugar in a reductive amination reaction; more preferably Z will be a
35 glycityl. Suitable reducing sugars include glucose, fructose, maltose, lactose,
galactose, mannose, and xylose. As raw materials, high dextrose corn syrup, highfructose corn syrup, and high maltose corn syrup can be utilized as well as the
... ., . . . .. .... , .. ~ . . ~ .
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26
individual sugars listed above. These corn syrups may yield a mix of sugar
components for Z. It should be understood that it is by no means intended to
exclude other suitable raw materials. Z preferably will be selected from the group
consisting of-CH2-(CHOH)n-CH2OH, -CH(CH2OH)-(CHOH)n l-CH2OH, -CH2-
5 (CHOH)2(CHOR')(CHOH)-CH2OH, and alkoxylated derivatives thereof, where n is
an integer from 3 to 5, inclusive, and R' is H or a cyclic or aliphatic monosaccharide.
Most preferred are glycityls wherein n is 4, particularly -CH2-(CHOH)4-CH2OH.
R can be, for exarnple, N-methyl, N-ethyl, N-propyl, N-isopropyl, N-butyl, N-
2-hydroxy ethyl, or N-2-hydroxy propyl.
1 o R2-CO-NC can be, for example, cocamide, stearamide, oleamide, lauramide,
myristamide, capricamide, palmitamide, tallowamide, etc.
Z can be l-deoxyglucityl, 2-deoxyfructityl, I-deoxymaltityl, I-deoxylactityl, 1-deoxygalactityl, 1-deoxymannityl, l-deoxymaltotriotityl, etc.
Methods for m~ing polyhydroxy fatty acid amides are known in the art. In
general, they can be made by reacting an alkyl amine with a reducing sugar in a
reductive amination reaction to form a corresponding N-alkyl polyhydroxyamine,
and then reacting the N-alkyl polyhydroxyamine with a fatty aliphatic ester or
triglyceride in a conflen~tion/arnidation step to form the N-alkyl, N-polyhydroxy
fatty acid amide product. Processes for making compositions cont~ining
polyhydroxy fatty acid amides are disclosed, for exarnple, in G.B. Patent
Specification 809,060, published February 18, 1959, by Thomas Hedley & Co., Ltd.,
U.S. Patent 2,965,576, issued December 20, 1960 to E. R. Wilson, and U.S. Patent2,703,798, Anthony M. Schwartz, issued March 8, 1955, and U.S. Patent 1,985,424,issued December 25, 1934 to Piggott, each of which is incorporated herein by
reference.
Second Enzvme - ~efe~ d compositions herein further comprise a
performance-enhancing amount of a detergent-compatible second enzyme. By
"d~lelgelll-compatible" is meant compatibility with the other ingredients of a liquid
detergent composition, such as detersive surfactant and detergency builder. These
second enzymes are preferably selected from the group con~i~ting of lipase, amylase,
cellulase, and mixtures thereof. The term "second enzyme" excludes the proteolytic
enzymes discussed above, so each composition which has a second ert7yme containsat least two kinds of enzyme, including at least one proteolytic enzyme. The amount
of second enzyme used in the composition varies according to the type of enzyme.In general, from about 0.0001 to 0.3, more preferably 0.001 to 0.1, weight % of
these second enzymes are preferably used. Mixtures of the same class of enzymes
.. .... ... . . . ... . . .
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27
(e.g. Iipase) or two or more classes (e.g. cellulase and lipase) may be used. Purified
or non-purified forms of the enzyme may be used.
Any lipolytic enzyme suitable for use in a liquid detergent composition can be
used in these compositions. Suitable lipase enzymes for use herein include those of
bacterial and fungal origin.
Suitable bacterial lipases include those produced by microorg~nicm~ of the
Pseudomonas groups, such as Pseudomonas stutzeri ATCC 19.154, as disclosed in
British Patent 1,372,034, incorporated herein by reference. Suitable lipases include
those which show a positive immunological cross-reaction with the antibody of the
lipase produced by the microorganism Pseudomonas fluorescens IAM 1057. This
lipase and a method for its purification have been described in J~p~nPse Patent
Application 53-20487, laid open on February 24, 1978. This lipase is available from
Arnano Pharmaceutical Co. Ltd., Nagoya, Japan, under the trade name Lipase P
"Amano," hereinafter referred to as "Amano-P." Such lipases should show a
positive immunological cross-reaction with the Amano-P antibody, using the
standard and well-known immunodiffusion procedure according to Ouchterlony
(Acta. Med. Scan., 133, pages 76-79 (1950)). These lipases, and a method for their
immunological cross-reaction with Amano-P, are also described in U.S. Patent
4,707,291, Thom et al., issued November 17, 1987, incorporated herein by reference.
Typical examples thereof are the Amano-P lipase, the lipase ex Pseudomonas fra iFERM P 1339 (available under the trade name Amano-B), lipase ex Pseudomonas
nitroreducens var. Iipolyticum FERM P 1338 (available under the trade name
Amano-CES), lipases ex Chromobacter viscosum, e.g. Chromobacter viscosum var.
Iipolvticum NRRLB 3673, commercially available from Toyo Jozo Co., Tagata,
Japan; and further Chromobacter viscosum lipases from U.S. Biochemical Corp.,
U.S.A. and Disoynth Co., The Netherlands, and lipases ex Pseudomonas ~ladioli.
Suitable fungal lipases include those producible by Hu~nicola lanuginosa and
Thermomyces lanu~inosus. Most preferred is lipase obtained by cloning the gene
from Humicola lanu~inosa and ~ressing the gene in Asper~illus orvzae as
described in European Patent Application 0 258 068 (Novo Industri A/S),
commercially available from Novo Nordisk A/S under the trade name Lipolase(~.
From about 10 to 18,000, preferably about 60 to 6,000, lipase units per grarn
(LU/g) of lipase can be used in these compositions. A lipase unit is that amount of
Iipase which produces I mmol of titratable fatty acid per minute in a pH stat, where
pH is 9.0, tt;lllp~ldlu~e is 30~C, substrate is an emulsion of 3.3wt % of olive oil and
3.3% gurn arabic, in the presence of 13 mmol/l Ca and 20 rnmol/l NaCI in 5
mmol/l Tris-buffer.
, . . , . ~ ,,
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28
Any cellulase suitable for use in a liquid detergent composition can be used in
these compositions. Suitable cellulase enzymes for use herein include those frombacterial and fungal origins. Preferably, they will have a pH optimum of between 5
and 9.5. From about 0.0001 to 0.1 weight % cellulase can be used.
Suitable cellulases are disclosed in U.S. Patent 4,435,307, Barbesgaard et al.,
issued March 6, 1984, incorporated herein by reference, which discloses fungal
cellulase produced from Humicola insolens. Suitable cellulases are also disclosed in
GB-A-2.075.028, GB-A-2.095.275 and DE-OS-2.247.832.
Examples of such cellulases are cellulases produced by a strain of Humicola
1 o insolens (Humicola ~risea var. therrnoidea), particularly the Humicola strain DSM
1800, and cellulases produced by a fungus of Bacillus N or a cellulase 212-
producing fungus belonging to the genus Aeromonas, and cellulase extracted from
the hepatopancreas of a marine mollusc (Dolabella Auricula Solander).
Any arnylase suitable for use in a liquid detergent composition can be used in
15 these compositions. Amylases include, for exarnple, amylases obtained from a
special strain of B.licheniformis, described in more detail in British Patent
Specification No. 1,296,839 (Novo). Amylolytic proteins include, for exarnple,
RapidaseR, International Bio-Synthetics, Inc. and TermamylR Novo Industries.
From about 0.0001% to 0.55, preferably 0.0005 to 0.1, w~. % amylase can be
20 used.
Optional In~redients - Detergent builders can optionally be included in the
compositions herein, especially for laundry compositions. Inorganic as well as
organic builders can be used. When present, the compositions will typically
comprise at least about 1 % builder and can be either an inorganic or organic builder.
25 Liquid laundry formulations preferably comprise from about 3% to 30%, more
preferably about 5 to 20%, by weight, of detergent builder.
Inorganic detergent builders include, but are not limited to, the alkali metal,
ammonium and alkanolarnmonium salts of polyphosphates (exemplified by the
tripolyphosph~es, pyrophosphates, and glassy polymeric meta-phosphates),
30 phosphonates, phytic acid, silicates, carbonates (including bicarbonates and
sesquicarbonates), sulphates, and alnminosilicates. Borate builders, as well as
builders cont~ining borate-forrning materials that can produce borate under detergent
storage or wash conditions (hereinafter, collectively "borate builders"), can also be
used. Preferably, non-borate builders are used in the compositions of the invention
35 intPndecl for use at wash conditions less than about 50~C, especially less than about
40~C.
, .. ...
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29
Examples of silicate builders are the alkali metal silicates, particularly thosehaving a SiO2:Na20 ratio in the range 1.6:1 to 3.2:1 and layered silicates, such as
the layered sodium silicates described in U.S. Patent 4,664,839, issued May 12,
1987 to H. P. Rieck, incorporated herein by reference. However, other silicates may
5 also be useful such as for example m~gnesiurn silicate, which can serve as a
cricpening agent in granular formulations, as a stabilizing agent for oxygen bleaches,
and as a component of suds control systems.
Examples of carbonate builders are the Alk~line earth and alkali metal
carbonates, including sodium carbonate and sesquicarbonate and mixtures thereof
10 with ultra-fine calcium carbonate as disclosed in German Patent Application No.
2,321,001 published on November l S, 1973, the disclosure of which is incorporated
herein by reference.
Aluminosilicate builders are useful in the present invention. Aluminosilicate
builders are of great importance in most currently marketed heavy duty granular
5 detergent compositions, and can also be a significant builder ingredient in liquid
detergent formulations. Aluminosilicate builders include those having the empirical
formula:
MZ(zAlo2 YSiO2)
wherein M is sodiurn, potassium, ammonium or substituted ammonium, z is from
20 about 0.5 to about 2; and y is 1; this material having a m~gn.-sium ion exchange
capacity of at least about SO milligram equivalents of CaC03 hardness per gram of
anhydrous aluminosilicate. Preferred alumino-silicates are zeolite builders which
have the formula:
Naz[(A102)z (sio2)yJ-xH2o
25 wherein z and y are integers of at least 6, the molar ratio of z to y is in the range
from 1.0 to about 0.5, and x is an integer from about lS to about 264.
Useful aluminosilicate ion exchange materials are commercially available.
These aluminosilicates can be crystalline or amorphous in structure and can be
naturally-occ~lrring aluminosilicates or synthetically derived. A method for
30 producing aluminosilicate ion exchange materials is disclosed in U.S. Patent
3,985,669, Krummel, et al., issued October 12, 1976, incorporated herein by
reference. Preferred synthetic crystalline aluminosilicate ion exchange materials
useful herein are available under the desi~n~tions Zeolite A, Zeolite P ~B), andZeolite X. In an especially preferred embodiment, the crystalline aluminosilicate ion
35 exchange material has the formula:
Nal2[(Alo2)l2(sio2)l2] XH20
~ . . .. . . . .
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wherein x is from about 20 to about 30, especially about 27. This material is known
as Zeolite A. Preferably, the aluminosilicate has a particle size of about 0.1 - 10
microns in diameter.
Specific exarnples of polyphosphates are the alkali metal tripolyphosphates,
sodium, potassium and ammonium pyrophosphate, sodium and potassium and
ammonium pyrophosphate, sodium and potassium orthophosphate, sodium polymeta
phosphate in which the degree of polymerization ranges from about 6 to about 21,and salts of phytic acid.
Examples of phosphonate builder salts are the water-soluble salts of ethane I -
10 hydroxy- 1, 1 -diphosphonate particularly the sodium and potassium salts, the water-
soluble salts of methylene diphosphonic acid e.g. the trisodium and tripotassiumsalts and the water-soluble salts of substituted methylene diphosphonic acids, such
as the trisodium and tripotassium ethylidene, isopyropylidene benzylmethylidene
and halo methylidene phosphonates. Phosphonate builder salts of the
aforementioned types are disclosed in U.S. Patent Nos. 3,159,581 and 3,213,030
issued December 1, 1964 and October 19, 1965, to Diehl; U.S. Patent No. 3,422,021
issued January 14, 1969, to Roy; and U.S. Patent Nos. 3,400,148 and 3,422,137
issued September 3, 1968, and January 14, 1969 to Quimby, said disclosures beingincorporated herein by reference.
Organic detergent builders preferred for the purposes of the present invention
include a wide variety of polycarboxylate compounds. As used herein,
"polycarboxylate" refers to compounds having a plurality of carboxylate groups,
preferably at least 3 carboxylates.
Polycarboxylate builder can generally be added to the composition in acid
25 form, but can also be added in the form of a neutralized salt. When utilized in salt
form, alkali metals, such as sodiurn, potassium, and lithiurn, or alkanolarnmonium
salts are plefe.lcd.
Included arnong the polycarboxylate builders are a variety of categories of
useful materials. One important category of polycarboxylate builders encomp~cses30 the ether polycarboxylates. A number of ether polycarboxylates have been disclosed
for use as detergent builders. Examples of useful ether polycarboxylates includeoxydisuccinate, as disclosed in Berg, U.S. Patent 3,128,287, issued April 7, 1964,
and Lamberti et al., U.S. Patent 3,635,830, issued January 18, 1972, both of which
are incol~oldled herein by reference.
A specific type of ether polycarboxylates useful as builders in the present
invention also include those having the general formula:
CH(A)(COOX)-CH(COOX)-O-CH(COOX)-CH(COOX)(B)
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31
wherein A is H or OH; B is H or -O-CH(COOX)-CH2(COOX); and X is H or a salt-
forming cation. For exarnple, if in the above general formula A and B are both H,
then the compound is oxydissuccinic acid and its water-soluble salts. If A is OH and
B is H, then the compound is tartrate monosuccinic acid (TMS) and its water-soluble
salts. If A is H and B is -O-CH(COOX)-CH2(COOX), then the compound is tartrate
disuccinic acid (TDS) and its water-soluble salts. Mixtures of these builders are
especially preferred for use herein. Particularly preferred are mixtures of TMS and
TDS in a weight ratio of TMS to TDS of from about 97:3 to about 20:80. These
builders are disclosed in U.S. Patent 4,663,071, issued to Bush et al., on May 5,
O 1987.
Suitable ether polycarboxylates also include cyclic compounds, particularly
alicyclic compounds, such as those described in U.S. Patents 3,923,679; 3,835,163;
4,158,635; 4,120,874 and 4,102,903, all of which are incorporated herein by
reference.
Other useful detergency builders include the ether hydroxypolycarboxylates
represented by the structure:
HO-[C(R)(COOM)-C(R)(COOM)-O]n-H
wherein M is hydrogen or a cation wherein the resultant salt is water-soluble,
preferably an alkali metal, ammonium or substituted ammonium cation, n is from
about 2 to about 15 (preferably n is from about 2 to about 10, more preferably naverages from about 2 to about 4) and each R is the same or different and selected
from hydrogen, C I 4 alkyl or C I 4 substituted alkyl (preferably R is hydrogen).
Still other ether polycarboxylates include copolymers of maleic anhydride with
ethylene or vinyl methyl ether, 1, 3, 5-trihydroxy benzene-2, 4, 6-trisulphonic acid,
and carboxymethyloxysuccinic acid.
Organic polycarboxylate builders also include the various alkali metal,
ammoniurn and substituted ammonium salts of polyacetic acids. Examples include
the sodium, potassium, lithium, ammonium and substituted arnmoniurn salts of
ethylen~ mine tetraacetic acid, and nitrilotriacetic acid.
Also included are polycarboxylates such as mellitic acid, succinic acid,
oxydisuccinic acid, polymaleic acid, benzene 1,3,5-tricarboxylic acid, and
carboxymethyloxysuccinic acid, and soluble salts thereof.
Citrate builders, e.g., citric acid and soluble salts thereof (particularly sodium
salt), are polycarboxylate builders of particular importance for heavy duty liquid
detergent formulations, but can also be used in granular compositions.
. ~._.. .. .. . .
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32
Other carboxylate builders include the carboxylated carbohydrates disclosed in
U.S. Patent 3,723,322, Diehl, issued March 28, 1973, incorporated herein by
reference.
Also suitable in the detergent compositions of the present invention are the 3,3-
dicarboxy-4-oxa-1,6-hexanedioates and the related compounds disclosed in U.S.
Patent 4,566,984, Bush, issued January 28, 1986, incorporated herein by reference.
Useful succinic acid builders include the Cs-C20 alkyl succinic acids and salts
thereof. A particularly preferred compound of this type is dodecenylsuccinic acid.
Alkyl succinic acids typically are of the general formula
o R-CH(COOH)CH2(COOH) i.e., derivatives of succinic acid, wherein R is
hydrocarbon, e.g., Clo-C20 alkyl or alkenyl, preferably C12-C16 or wherein R maybe substituted with hydroxyl, sulfo, sulfoxy or sulfone substituents, all as described
in the above-mentioned patents.
The succinate builders are preferably used in the form of their water-soluble
15 salts, including the sodium, potassium, ammonium and alkanolammonium salts.
Specific examples of succinate builders include: laurylsuccinate,
myristylsuccinate, palmitylsuccinate, 2-dodecenylsuccinate (plere.led), 2-
pentadecenylsuccinate, and the like. Laurylsuccinates are the preferred builders of
this group, and are described in European Patent Application 86200690.5/0,200,263,
20 published November 5, 1986.
Examples of useful builders also include sodium and potassiurn
carboxymethyloxymalonate, carboxymethyloxysuccinate, cis-cyclo-hexane-
hexacarboxylate, cis-cyclope~ e-tetracarboxylate, water-soluble polyacrylates
(these polyacrylates having molecular weights to above about 2,000 can also be
25 effectively utilized as disl)e~s~.l~), and the copolymers of maleic anhydride with
vinyl methyl ether or ethylene.
Other suitable polycarboxylates are the polyacetal carboxylates disclosed in
U.S. Patent 4,144,226, Crutchfield et al., issued March 13, 1979, incolyoldled herein
by reference. These polyacetal carboxylates can be ylepa~ed by bringing together,
30 under polymerization conditions, an ester of glyoxylic acid and a polymerization
initiator. The resulting polyacetal carboxylate ester is then attached to chemically
stable end groups to stabilize the polyacetal carboxylate against rapid
depolymerization in ~ lin-o solution, converted to the coll~;syo~lding salt, and added
to a surfactant.
Polycarboxylate builders are also disclosed in U.S. Patent 3,30~,067, Diehl,
issued March 7, 1967, incorporated herein by reference. Such materials include the
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33
water-soluble salts of homo- and copolymers of aliphatic carboxylic acids such as
maleic acid, itaconic acid and methylenemalonic acid.
Other organic builders known in the art can also be used. For example,
monocarboxylic acids, and soluble salts thereof, having long chain hydrocarbyls can
5 be lltili7~d These would include materials generally referred to as "soaps." Chain
lengths of C 1 0-C20 are typically utili7ec~ The hydrocarbyls can be saturated or
unsaturated.
Other optional ingredients include soil release agents, chelating agents, clay
soil removal/anti redeposition agents, polymeric dispersing agents, bleaches,
10 brl~htençrs, suds suppresors, solvents and aesthetic agents.
The detergent composition herein can be formulated as a variety of
compositions, for instance as laundry detergents as well as hard surface cleaners or
dishwashing compositions.
The compositions according to the present invention are further illustrated by
the following exarnples.
EXAMPLE I
The following compositions are made by combining the listed ingredients in
the listed plol,ol~ions. In this example, one or more of the following peptide
aldehydes are used:
20 Peptidealdehyde l: CH3O-(O)C-Phe-Gly-Ala-LeuH
Peptide aldehyde 2: CH3N-(O)C-Phe-Gly-Ala-LeuH
Peptide aldehyde 3: CH3O-(O)C-Phe-Gly-Ala-PheH
Peptide aldehyde 4: CH3N-(O)C-Phe-Gly-Ala-PheH
Peptidealdehyde5: CH3SO2Phe-Gly-Ala-Leu-H
25 Peptide aldehyde 6: CH3SO2Val-Ala-Leu-H
Peptide aldehyde 7: C6HsCH2O(OH)(O)P-Val-Ala-Leu-H
Peptidealdehyde8: CH3CH2SO2-Phe-Gly-Ala-Leu-H
Peptide aldehyde 9: C6HsCH2SO2-Val-Ala-Leu-H
Peptide aldehyde 10: C6HsCH2O(OH)(O)P-Leu-Ala-Leu-H
30 Peptidealdehyde 11: C6HsCH2O(OH)(O)P-Phe-Ala-Leu-H
Peptide aldehyde 12: CH3O(OH)(O)P-Leu-Gly-Ala-Leu-H.
Compositions A B C D E F
Linearalkyl benzene 8.5 15 6.5 10 12.5 4
sulfonic acid
Sodium C 12-15 1 2 l 2 -- --
alkyl sulfate
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34
C14 1salkyl2.5times10 5 105 11 9
ethoxylated sulfate
C12 glucose amide -- -- 9 -- 5
C12 1salcohol7times 3 10 4 7 2.5
ethoxylated
Fatty acid 2 5 5 4 2 2
Citricacid 6 7 4 6 4 5
C12 14alkenyl -- 6 -- 5 6
substituted, succinic acid
Sodium hydroxide 2 6 2 4 1 1.5
Ethanol 2 1.5 2 4 2 1.5
Monoethanolamine 6 5 4 --
1,2-Propanediol 12 10 5 5 4 6
Amylase (143 KNU/g) -- -- 0.1 0.2
Lipolase~) (lOOKLU/g 0.5 0.2 0.5 0.5 0.4 --
commercial solution)
ProteaseB (34g/L 0.9 -- 0.5 -- 1.2 --
commerical solution)
Savinase(~ -- 0.3 -- 0.4 0.2 0.3
(commercial solution)
Carezymeg) 0.5 1 0.8 -- 0.2 0.8
Peptide aldehydes 1-12 0.009 0.005 0.001 0.0005 0.0011 0.1
Calcium Ions 0.01 0.5 0.1 0.05 0.9 0.25
Water and minors Balance to 100%
EXAMPLE II
The following for nula is tested for % of protease activity rem~ining.
Combinations of 0%, 0.1%, 0.2%, and 0.3% Ca++ (from CaC12) and 0%, 0.0006%,
0.00125%, and 0.0025% peptide aldehyde (Synthesis Example 6) are used. Products
are held at 90~F and assayed at weekly intervals for 42 days.
Component Wt (%)
Alkyl, 1.4 ethoxylated, sulfate 30
Amine oxide 6
Polyhydroxy fatty acid amide 4
Nonionic surfactant (C 1 1 E9) 5
Mg ion from MgC12
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Ca ion from CaC12 see chart below
Peptide aldehyde* see chart below
Sodium xylene sulfonate 4
Solvent 6
Water to 100%
pH to 8
*Peptide Aldehydes of Synthesis Exarnple 6.
EXAMPLE III
Results showing the petcent protease activity re~n~ining after 42 days at 90~F.
0.01% Protease B enzyme is used.
Calcium Ion Peptide AldehYde
-- 0% 0.0006% 0.00125% 0.0025%
0% 48 71 75 86
0.1% 52 84 92 87
0.2% 53 85 98 100
0.3% 60 80 98 92
EXAMPLE IV
The following compositions are made by combining the listed ingredients in
the listed ~.opol lions.
In~redients A (wt %) B (wt %) C (wt %) D (wt %)
LAS 0 0 0 12
AExS I 22.1 24.7 33.5 3
Polyhydroxyfatty 4.6 1.2 4.2 0
acid amide
Amine Oxide 4.6 1.2 4.8 0
Betaine 0 1.2 0 0
Nonionic 6.7 4.1 0 0
Surfactant
Mg(OH)2 0.5 0.5 0.7 0
Ca ion from CaC12 0.1 0.3 0.4 0.1
Calciurn xylene 4.5 0 4 0
sulfonate
Polyethylene 3 ~ ~ ~
glycol
.. .. ~ .. ~
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36
Polypropylene 1.5 0 0 0
glycol 2000
Balance, water to 100% to 100% to 100% to 100%
Protease A or 0.001 -0.01 0.001 -0.01 0.005-0.01 0.0003-0.01
Protease B
Peptide 0.00025- 0.00025- 0.00025- 0.00125-
Aldehydes2 0.0025 0.0025 0.0025 0.0025
1 x= the degree of ethoxylation. The average degree of ethoxylation for the
compositions are: A=2.2, B=0.6, C=1.4, D=2.2.
2 The peptide aldehydes of Synthesis Example 6 are used herein.
