EP0929639B1

Movatterモバイル変換

Info

Publication number: EP0929639B1
Application number: EP97942579A
Authority: EP
Inventors: John Mcmillan Mciver; Alan Carl Huber; Kirsten Louise Mckillop; Laurence Anthony Smith
Original assignee: Procter and Gamble Co
Current assignee: Procter and Gamble Co
Priority date: 1996-09-24
Filing date: 1997-09-19
Publication date: 2002-11-13
Anticipated expiration: 2017-09-19
Also published as: DE69717133T2; JP2000506930A; ATE227769T1; AR009816A1; DE69717133D1; CN1238001A; EP0929639A1; CA2266527A1; WO1998013459A1

Description

TECHNICAL FIELD

This invention relates to liquid detergent compositions containing enzymes.More specifically, this invention pertains to liquid detergent compositionscontaining a detersive surfactant, a proteolytic enzyme, a peptide aldehyde, andcalcium ions. The combination of peptide aldehyde and calcium ions act to providesynergistic protease inhibitor benefits.

BACKGROUND OF THE INVENTION

Protease-containing liquid aqueous detergents are well-known, especially inthe context of laundry washing. A commonly encountered problem in suchprotease-containing liquid aqueous detergents is the degradation phenomenon by theproteolytic 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 secondenzyme or the protease itself in the detergent composition is affected and thedetergent composition consequently performs less well.
In response to this problem, it has been proposed to use various proteaseinhibitors or stabilizers. For instance, various references have proposed the use ofthe following compounds to aid in the stabilization of enzymes: benzamidinehydrochloride, lower aliphatic alcohols or carboxylic acids, mixtures of a polyol anda boron compound, aromatic borate esters, and calcium, particularly calciumformate. Recently, it was discovered that certain peptide aldehydes act to stabilizeprotease enzyme.
Although these compounds have been used to varying success in liquiddetergents, they are not free of problems. For example peptide aldehydes are ratherexpensive and create complexities for the formulators, especially for liquiddetergents. Other inhibitors such as calcium and boric acids are less expensive butdo not stabilize enzymes as well as peptide aldehydes. It is thus an object of thepresent invention to provide a protease inhibitor system which is economical,effective and suitable for use in a liquid detergent composition.
In response to this object, the present invention proposes to use a combinationof calcium ions and peptide aldehydes as reversible protease inhibitors in aqueous liquid detergent compositions. The presence of both calcium and peptide aldehydeprovides a synergistic stabilization of the protease. This novel combination providesthe formulator added flexibility in designing a stabilization system. The levels ofpeptide aldehyde and calcium can be adjusted to deliver the most cost effectiveformula and to minimize product stability problems that often arise from thepresence of divalent ions in a liquid detergent matrix.
In particular, the present invention allows for the use of very low levels ofpeptide aldehydes in the liquid detergent compositions herein. This is particularlycritical in the formulation of relatively inexpensive, concentrated liquid detergentcompositions which are encompassed by the present invention.
Because the combination of calcium and peptide aldehydes are so efficient ininhibiting proteases, another advantage of the present invention is that even enzymeswhich are highly sensitive to proteolytic degradation can now be incorporated inliquid detergent compositions comprising a protease. Moreover, it has also beendiscovered that the increased stability of the protease enzyme allows for improvedskincare benefits. These benefits include softening of the skin and hands and lessdrying from exposure of the hands to the dishwashing liquor.

BACKGROUND ART

It has been proposed to use various protease inhibitors or stabilizers. Forinstance, US 4,566,985 proposes to use benzamidine hydrochloride; EP 376 705proposes to use lower aliphatic alcohols or carboxylic acids; EP 381 262 proposes touse 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. Alsosee US4,261,868; US4,404,115; US4,318,818; and EP130,756.
The use of peptide derivatives for the inhibition of proteins appears to havebeen disclosed in therapeutic applications. EP 293 881 discloses the use of peptideboronic acids as inhibitors of trypsin-like serine proteases. EP 185 390 and US4,399,065 disclose the use of certain peptide aldehydes derivatives for the inhibitionof blood coagulation. J 90029670 discloses the use of optically active alpha aminoaldehydes for the inhibition of enzymes in general. See also "Inhibition ofThrombin and Trypsin by Tripeptide Aldehydes",Int. J. Peptide Protein Res., Vol12 (1978), pp. 217-221; Gaal, Bacsy & Rappay, and "Tripeptide Aldehyde ProteaseInhibitors May Depress in Vitro Prolactin and Growth Hormone Release"Endocrinology, Vol. 116, No. 4 (1985), pp. 1426-1432; Rappay, Makara, Bajusz &Nagy. Certain peptide aldehydes have also been disclosed in EP-A-473 502 forinhibiting protease-mediated skin irritation.
In particular see EP185,390, WO94/04651, published 3 March 1994,WO94/04652, 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,and US5,527,487.
JP 62 269 689 describes a method for stabilising enzymes in detergentcompositions by adding a reversible inhibitor for the enzymeand optionally adding calcium salt. WO 92/03529 disclosesa detergent composition comprising a protease and one ormore enzymes, as well as a reversible protease inhibitorof the peptide or protein type.

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;
c) a source of calcium ions; other than a compound of the formula RO(A)_mSO₃ M; R, A and M, m are described in claim 1 and
d) a peptide aldehyde having the formula:Z-B-NH-CH(R)-C(O)H

₂

₃

₂

₁

₆

₇

₉

₄

₈

₁

₆

₇

₉

Without being limited by theory, it is believed that the combined source ofcalcium ion and peptide aldehyde provides more than additive stability to theproteolytic enzyme.
Preferably, the liquid detergent compositions herein comprise, by weight ofcomposition:
a) from 1 to 95%, preferably from 8% to 70%, of saiddetersive surfactant;
b) from 0.0001% to 5%, preferably from 0.0003% to0.1%, of an active proteolytic enzyme;
c) from 0.00001% to 5%, preferably from 0.0001% to1%, more preferably from 0.0006% to 0.5%, of a peptide aldehyde asdescribed hereinbefore; and
d) from 0.01% to 1%, preferably from 0.05% to 0.5%, ofcalcium ion.
The proteolytic enzyme useful herein is preferably a subtilisin-type proteaseand 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 calciumformate, calcium xylene sulfonate, calcium chloride, calcium acetate, calciumsulfate. 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, andmixtures thereof. Moreover for dishcare compositions, the liquid detergentcompositions may further comprise an effective amount of amylase enzyme.Additionally, the dishcare compositions may optionally comprise an effectiveamount of a source of boric acid and a diol. Typically dishcare compositions willoptionally, but preferably, comprise from about 0.25% to about 10%, preferablyfrom about 0.5% to about 5%, more preferably from about 0.75% to about 3%, byweight of boric acid or a compound capable of forming boric acid and a diol, e.g.1,2-propanediol.
In a preferred embodiment for heavy duty detergent compositions useful inlaundry care, the liquid detergent composition further comprises an effective amountone or more of the following enzymes: lipase, amylase, cellulase, and mixturesthereof. Preferably for laundry compositions, the second enzyme is lipase and isobtained by cloning the gene fromHumicola Lanuginosa and expressing the gene inAspergillus Oryzae. Lipase is utilized in an amount of from about 10 to about 18000lipase units per gram, preferably from about from about 60 to about 6000 units pergram.
In another preferred composition useful for laundry care, the second enzyme isa cellulase derived fromHumicola Insolens and is utilized in an amount of fromabout 0.0001% to about 0.1% by weight of the total composition of said cellulase.
The compositions herein may contain further detersive adjuncts, including butnot limited to, one or more of the following: suds boosters, builders, soil releasepolymers, polyacrylate polymers, dispersing agents, dye transfer inhibitors, dyes,perfumes, processing aids, brighteners, and mixtures thereof. Additionally, forlaundrycare compositions, the detersive surfactant is typically present in an amountof from 10% to 70%, by weight of total composition. Moreover, thelaundry compositions may optionally comprise an effective amount of a source of boric acid and a diol. Typically laundry compositions will optionally, butpreferably, comprise from about 0.25% to about 10%, preferably from about 0.5% toabout 5%, more preferably from about 0.75% to about 3%, by weight of boric acidor a compound capable of forming boric acid and a diol, e.g. 1,2-propanediol.
All percentages and proportions herein are by weight, and all references citedare hereby incorporated by reference, unless otherwise specifically indicated.

DETAILED DESCRIPTION OF THE INVENTION

Definitions - The present detergent compositions comprise an "effectiveamount" or a "stain removal-improving amount" of individual components definedherein. An "effective amount" 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 enzymestability benefit when the calcium and peptide aldehydes are combined is greaterthan the sum of the individual benefits obtained when only one of the components ispresent in a detergent composition.
The liquid aqueous detergent compositions according to the present inventioncomprise 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 furthercomprise (E) a detergent-compatible second enzyme or a mixture thereof, and mayfurther comprise (F) other optional ingredients.
Peptide aldehydes - The detergent compositions according to the presentinvention comprise, as a first essential ingredient, a peptide aldehyde having theformula:Z-B-NH-CH(R)-C(O)H wherein B is a peptide chain comprising from 1 to 5 amino acid moieties; Zis an N-capping moiety selected from the group consisting of phosphoramidate[(R"O)₂(O)P-], 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 orbranched C₁-C₆ unsubstituted alkyl, phenyl, C₇-C₉ alkylaryl, and cycloalkylmoieties, wherein the cycloalkyl ring may span C₄-C₈ and may contain one or moreheteroatoms selected from the group consisting of O,N,and S (preferred R" is selected from the group consisting of methyl, ethyl, and benzyl); and R is selectedfrom the group consisting of straight or branched C₁ - C₆ unsubstituted alkyl,phenyl, and C₇ - C₉ alkylaryl moieties.
Preferred R moieties are selected from the group consisting of methyl, iso-propyl,sec-butyl, iso-butyl, -C₆H₅, -CH₂-C₆H₅, and -CH₂CH₂-C₆H₅, whichrespectively may be derived from the amino acids Ala, Val, Ile, Leu, PGly(phenylglycine), Phe, and HPhe (homophenylalanine) by converting the carboxylicacid group to an aldehyde group. While such moieties are therefore not amino acids(and they may or may not have been synthesized from an amino acid precursor), forpurposes of simplification of the exemplification of inhibitors useful here, thealdehyde portion of the inhibitors are indicated as derived from amino acids by theaddition of "H" after the analogous amino acid [e.g., "-AlaH" represents thechemical moiety "-NHCH(CH₃)C(O)H"].
Preferred B peptide chains are selected from the group consisting of peptidechains having the amino acid sequences according to the general formula:Z-A⁵-A⁴-A³-A²-A¹-NH-CH(R)-C(O)Hsuch that the following amino acids, when present, are :
A¹ 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 correspondingamino acid whereby the C-terminal end of said amino acid is converted from acarboxylic group to an aldehyde group. Such aldehydes may be prepared by knownprocesses, for instance as described in US 5015627, EP 185 390, EP 583,534, andDE 32 00 812.
While not wanting to be bound by theory it is believed that the peptidealdehydes according to the present invention bind to the proteolytic enzyme in theliquid detergent composition, thereby inhibiting said proteolytic enzyme. Upondilution in water, the proteolytic activity is restored by dissociation of the proteolyticenzyme/peptide aldehyde complex.
The N-terminal end of said protease inhibitors according to the presentinvention is protected by one of the N-capping moiety protecting groups selectedfrom the group consisting of carbamates, ureas, sulfonamides,phosphonamides,thioureas, sulfenamides, sulfonic acids, phosphinamides,thiocarbamates, amidophosphates, and phosphonamides. However, in a highly preferred embodiment of the present invention, the N-terminal end of said proteaseinhibitor is protected by a methyl, ethyl or benzyl carbamate [CH₃O-(O)C-;CH₃CH₂O-(O)C-; or C₆H₅CH₂O-(O)C-], methyl, ethyl or benzyl urea [CH₃NH-(O)C-;CH₃CH₂NH-(O)C-; or C₆H₅CH₂NH-(O)C-], methyl, ethyl or benzylsulfonamide [CH₃SO₂-; CH₃CH₂SO₂-; or C₆H₅CH₂SO₂-], and methyl, ethyl orbenzyl amidophosphate [CH₃O(OH)(O)P-; CH₃CH₂O(OH)(O)P-; orC₆H₅CH₂O(OH)(O)P-] groups.
Synthesis of N-capping groups can be found in the following references:Protective Groups in Organic Chemistry, Greene, T., Wuts, P., John Wiley & Sons,New York, 1991, pp 309-405; March, J,Advanced Organic Chemistry, WileyInterscience, 1985, pp. 445, 469, Carey, F. Sundberg, R.,Advanced OrganicChemistry, Part B, Plenum Press, New York, 1990, pp. 686-89; Atherton, E.,Sheppard, R.,Solid Phase Peptide Synthesis, Pierce Chemical, 1989, pp. 3-4; Grant,G.,Synthetic Peptides, W. H. Freeman & Co. 1992, pp. 77-103; Stewart, J., Young,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: CH₃SO₂Phe-Gly-Ala-Leu-H,CH₃SO₂Val-Ala-Leu-H, C₆H₅CH₂O(OHXO)P-Val-Ala-Leu-H, CH₃CH₂SO₂-Phe-Gly-Ala-Leu-H,C₆H₅CH₂SO₂-Val-Ala-Leu-H, C₆H₅CH₂O(OH)(O)P-Leu-Ala-Leu-H,C₆H₅CH₂O(OH)(O)P-Phe-Ala-Leu-H, and CH₃O(OH)(O)P-Leu-Gly-Ala-Leu-H.
In the Synthesis Examples hereinafter methods are disclosed to synthesizecertain of these peptide aldehydes.

Synthesis Example 1

Synthesis of the tetrapeptide aldehyde Moc-Ala-Phe-Gly-Ala-LeuH

(a) Ala-Leu-OMe.HCL: To a solution of 3.0 g (14.83 mmol) Ala-Leu-OH, which isdissolved 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 andevaporated 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-OHand 0.500 g (1.98 mmol) Ala-Leu-OMe.HCl in CH₂Cl₂ is add 0.607 mlTEA followed immediately by 0.355 ml DEPC. The solution is stirred overnight,evaporated, and the residue partitioned between EtOAc and 1N HCl. The organic phase is washed successively with saturated NaHCO₃ 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 mmol) Ala-Phe which isdissolved in 4.23 ml 1N NaOH and cooled to 0°C, 0.419 g (4.44 mmol) is adedmethyl chloroformate dropwise. At the same time, in a separate addition funnel, anadditional 4.23 ml 1N NaOH is added such that the pH is maintained between 9.0-9.5.After addition is complete the reaction is stirred 30 minutes at 0°C and 2 h atroom temperature. At this point the solution is cooled to 0° and the pH adjusted to9.5. This basic solution is washed with EtOAc (1X, 100 ml). The aqueous (0°C) isthen 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 mmol) Cbz-Gly-Ala-Leucinemethyl ester in 10 ml MeOH is added 0.100 g 10% Pd/C. Thissolution is hydrogenated in the presence of 0.600 ml 4.0M HCl/Dioxane (underballoon pressure) for 1 h, filtered through celite and evaporated. This residue issuspended in CH₂Cl₂, 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 stirringovernight the solvent is evaporated, the residue partitioned between EtOAc and 1NHCl and washed successively with saturated NaHCO₃ and NaCl. 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 mmol) CaCl₂ in a mixture of 4 ml ethanol and 2 ml THE. This mixture iscooled to -15°C and 0.450 g (0.820 mmol) Moc-Ala-Phe-Gly-Ala-Leu-OMe isadded followed by 0.124 g (3.28 mmol) NaBH₄. The reaction is stirred for 2 h andquenched with 10 ml 1N HCl. The solvents are evaporated and the remainingaqueous layer partitioned with EtOAc. The organic phase is then washed withsaturated NaHCO₃ and saturated NaCI. Drying (MgSO₄), evaporation andchromatography affords 0.256 g of pure product.
(f) Moc-Ala-Phe-Gly-Ala-LeuH: A solution is prepared by adding 0.623 g (1.47mmol) Dess-Martin periodinane to 1.8 L CH₂Cl₂ followed by stirring for 10minutes. This solution is then cooled to 0°C and 0.256 g (0.490 mmol) Etoc-Phe-Gly-Ala-Leucinolis added in one portion. The reaction is continued for 2 h andpoured into a solution consisting of 2.55 g (10.47 mmol) Na₂S₂O₃ in 30 ml saturated NaHCO₃. After stirring for 10 minutes the mixture is extracted withEtOAc (2X, 50 ml). The combined extracts are dried (MgSO₄), evaporated, andchromatographed 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 isdissolved in 4.5 L of MeOH and cooled to 0°C, is added 178.6 ml (4.95 mol) ofthionyl chloride dropwise. The solution is stirred overnight at room temperature andevaporated to dryness providing 543 g (97.1% yield) of the desired product to beused as is.
(b) Etoc-Phe-Gly-OH: To a solution of 450 g (2.03 mol) Phe-Gly which isdissolved in 2026 ml 1N NaOH and cooled to 0°C, is added methyl chloroformate(3.1 ml, 40.0 mmol) dropwise. At the same time, in a separate addition funnel, anadditional 2026 ml 1N NaOH is added such that the pH is maintained between 9.0-9.5.After addition is complete the reaction is stirred 30 minutes at 0°C and 2 h atroom temperature. At this point the solution is cooled to 0° and the pH adjusted to9.5. This basic solution is washed with EtOAc (1X, 4 L). The aqueous (0°C) is thenadjusted to pH = 2.5 (2N HCl) 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-OHand 546 g (1.86 mol) Ala-Leu-OMe.HCl in 8 liters CH₂Cl₂ 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 washedconsecutively with 2 liters each of 2N HCl, sat'd NaHCO₃ and sat'd NaCI. Theorganic phase is then dried (MgSO₄), filtered and evaporated to yield 916 g (93%yield) of the desired material.
(d) Etoc-Phe-Gly-Ala-Leucinol: To a solution of 45.10 g (0.406 mol) CaCl₂ in 1 Lethanol and 1 L THF 100 g (0.203 mol) of Etoc-Phe-Gly-Ala-Leu-OMe is addedand the mixture cooled to -15°C. To this solution 30.7 g (0.812 mmol) NaBH₄ iscarefully added followed by stirring for 2 h. Subsequently the reaction is quenchedwith 100ml 0. IN HCl. This solution is transfered to 4 L of 1N HCl and extractedwith EtOAc (3X, 2.75 L). The combined EtOAc layers are washed with 4 L saturated NaHCO₃, dried (MgSO₄) and evaporated. Trituration (twice) with ether (4L) 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 CH₂Cl₂ followed by stirring for 10 minutes. Thissolution is then cooled to 0°C and 60 g (0.13 mol) Etoc-Phe-Gly-Ala-Leucinoladded in one portion. The reaction is continued for 105 minutes and poured into asolution 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(1.5 L each) with CH₂Cl₂ are performed. The combined extracts are dried (MgSO₄),evaporated, and triturated with (2X, 1L) ether to provide 51.7 g (86.2% yield) of theproduct.

Synthesis Example 3:

Synthesis 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 isdissolved in 50 ml of MeOH and cooled to 0°C, is added 2.43 ml (33.36 mmol) ofthionyl chloride dropwise. The solution is stirred overnight at room temperature andevaporated 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 mmol) Cbz-Gly-OHand 0.500 g (1.98 mmol) Ala-Leu-OMe.HCl in CH₂Cl₂ 0.607 ml TEA isadded followed immediately by 0.355 ml DEPC. The solution is stirred overnightand then evaporated. The residue is partitioned between EtOAc and 1N HCl, theorganic phase is washed with saturated NaHCO₃ and saturated NaCl, dried(MgSO₄) 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-Leucinemethyl ester which is dissolved in 20 ml MeOH is added 0.200 g10% Pd/C. This is hydrogenated in the presence of 2.45 ml (9.81 mmol) 4.0MHCl/Dioxane for 2 h after which the reaction is thoroughly outgassed and filteredhrough Celite to remove the catalyst Evaporation of the MeOH affords 1.45 g ofpure product which is suspended in 45 ml CH₂Cl₂ and cooled to 0°C. To thissolution 1.45 ml (3.25 mmol) TEA is added followed by 0.362 ml methylchloroformate. After stirring overnight the CH₂Cl₂ is evaporated and the residuepartitioned between EtOAc and 1N HCl. The organic phase is separated and washed sequentially with NaHCO₃ and NaCl. Drying, (MgSO₄), evaporation andchromatographic purification affords 0.820 g of desired product.
(d) Moc-Gly-Ala-Leucinol: To a solution of 0.168 g (1.51 mmol) CaCl₂ in 25 mlethanol and 15 ml THF is added 0.250 g Moc-Gly-Ala-Leucine methyl ester. Thissolution 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 1N HCl, concentrated onrotovape and extracted with EtOAc (2x 50ml). The combined extracts are washedwith saturated NaHCO₃ and NaCl, dried (MgSO₄) and evaporated. Purification onsilica provides 0.167 g of the pure product.
(e) Moc-Gly-Ala-LeuH- A solution is prepared by adding 0.418 g (0.989 mmol)Dess-Martin periodinane to 5 ml CH₂Cl₂ followed by stirring for 10 minutes. Next0.100 g (0.330 mmol) Moc-Gly-Ala-Leucinol is added in one portion and thereaction stirred for 2 h and poured into a 25 ml solution of saturated NaHCO₃containing 1.72 g (6.93 mmol) Na₂S₂O₃. After stirring an additional 10 minutes thesolution is extracted with EtOAc (3X, 50 ml), dried (MgSO₄) and evaporated.Chromatography on silica affords 0.016 g of the desired product.

Synthesis 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 isdissolved in 9 ml 1N NaOH and cooled to 0°C, is added simultaneously 0.766 ml (9.9 mmol) of methane sulfonyl chloride and 9 ml 1N NaOH, in separate additionfunnels. After addition is complete the reaction is stirred 15 minutes at 0°C and 1 hat room temperature. At this point the solution is cooled to 0°C, the pH adjusted to9.5 and is washed with EtOAc (1X, 50 ml). The aqueous phase (0°C) is thenadjusted to pH = 2.5 (2N HCl) 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 0.366ml (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.HCl, in amixture of 10 ml THF and minimal DMF, are added. Stirring is continued at 0°C for15 minutes and 2 h at room temperature. The solution is quenched with 5 ml 1NHCl, extracted with EtOAc (3X, 50 ml), the combined extracts are washed with sat'd NaHCO₃ and sat'd NaCl. The resulting organic phase is then dried (MgSO₄),filtered, evaporated and chromatographed on silica to yield 0.260 g of the desiredmaterial.
(c) N-Ms-Phe-Gly-Ala-LeuH: A solution is prepared by adding 0.337 g (0.798mmol) Dess-Martin periodinane to 5 ml CH₂Cl₂ and stirring for 10 minutes. To thissolution 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 containing 1.8 g (5.586 mmol)Na₂S₂O₃. After stirring for 10 minutes the mixture is extracted with EtOAc (3X, 50ml). The combined extracts are dried (MgSO₄), evaporated, and chromatographedon silica to afford 0.048 g of the product.

Synthesis Example 5:

Synthesis of an aldehyde protease inhibitor

Moc-Leu-OH-L-Leucine (5.0 g, 38.2 mmol) is dissolved in 38 ml 1N NaOH andcooled to 0°C. Methyl chloroformate (3.1 ml, 40.0 mmol) is added dropwise whilein a separate addition funnel 1N NaOH is added as to maintain pH at 9.0-9.5. Afteraddition is complete and the pH stabilized at 9.0-9.5 the solution is washed with 200ml EtOAc. the aqueous phase is then acidified to pH = 2. This mixture is extractedwith EtOAc (2X 100 ml), dried (MgSO₄), filtered, and the solvent removed to afford7.15 g pure product.
Moc-Leu-Leucinol- To a solution of 3.5 g (18.52 mmol) Moc-Leu-OH in 100 mlTHF, cooled to -15°C, 2.04 ml (18.52 mmol) of N-methyl morpholine is addedfollowed immediatedly by 2.4 ml (18.52 mmol) isobutyl chloroformate. Afterstirring for 10 minutes 2.37 ml (18.52 mmol) of leucinol in 25 ml of THF is addedand the reaction stirred 0.5 h at -15°C and 1 h at room temperature. The mixture isthen diluted with 100 ml of H₂O and the THF evaporated. The remaining aqueousphase is partitioned between EtOAc and 1N HCl, the organic phase washed withNaHCO₃, dried (MgSO₄) and evaporated to afford 5.33 g pure product.
Moc-Leu-LeuH-A solution containing 4.4 g (10.41 mmol) Dens-Martin periodinanesuspended in 100 ml CH₂Cl₂ is prepared and stirred for 10 minutes. To this solution1.0 g (3.47 mmol) Moc-Leu-Leucinol is added and the solution stirred 2 h at roomtemperature followed by pouring into 100 ml of saturated NaHCO₃ containing 18 g(72.87 mmol) Na₂S₂O₃. This solution is stirred 10 minutes and then extracted with EtOAc (2X, 125ml), dried (MgSO₄) and the solvent evaporated. Chromatographyon silica affords 0.550 g of pure product.

Synthesis Example 6:

Additional peptide aldehydes are synthesized according to the followingprocedures. Some of the intermediates are purchased from suppliers and in theseinstances it is noted within the procedure. Dess-Martin periodinane is synthesizedaccording to the procedure of Martin, J.Org. Chem.,1983, 48, 4155.
I. Z-Gly-Ala-Leu-OMe - To a solution of Z-Gly-Ala-OH (20.0 g, 0.071 M) andLeu-OMe.HCl (12.9 g, 0.071 M) in 250 ml dichloromethane is added 21.9 ml (0.157M) triethylamine (TEA) dropwise over a period of 10 min. This addition is followedby the addition of 11.9 ml (0.078 M) of diethylcyanophosphonate (DECP). Themixture is stirred overnight and the solvent removed. The residue is dissolved inethyl acetate and washed with 1N HCl, saturated NaHCO₃, and brine. The solutionis dried with MgSO₄, filtered and the solvent removed. Recovered will be 29.0 g ofproduct that is homogeneous by TLC.¹³C NMR (CDCl₃) 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) isdissolved in 300 ml MeOH and 35 ml 4.0 M HCl in dioxane. To this solventmixture is added 5.8 g of 10% Pd/C portionwise. The slurry is degassed with anaspirator and H₂ introduced via balloon. The slurry is maintained under a positivepressure of H₂ and stirred overnight. The slurry is filtered through Celite and asintered glass funnel and washed thoroughly with MeOH. The solvent is removedand the residue is triturated with ether. The slurry is filtered and the filter cake driedunder vacuum. Recovered 20.2 g of an off-white powder. The crude product andMoc-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 residueis dissolved in EtOAc and washed with 1N HCl, saturated NaHCO₃, and brine. Theorganic phase is dried (MgSO₄), filtered and the solvent removed to afford 21.3 gproduct.¹³C NMR (CDCl₃) 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.
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 cooledto 0°C and 9.88 g (89.0 mmol) CaCl₂ is added. In 5 min the slurry will behomogenized and 6.73 g (178.0 mmol) NaBH₄ added portionwise over a period of 5min. The solution is stirred at 0°C for 2 hours and the reaction carefully quenchedwith 1N HCl. The EtOH and THF are removed under vacuum and the remainingaqueous mixture extracted with 500 ml EtOAc. This organic phase is washed withsaturated NaHCO₃, brine, and the organic phase dried with MgSO₄. Filtration andremoval of solvent affords 20.0 g of an off-white crystalline material.
Chromatography on silica (3.5% MeOH/CH₂Cl₂) gives 13.0 g pure product R_f=0.3 (10% MeOH/CH₂Cl₂),¹³C NMR (CDCl₃) 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, 136.62, 157.85, 170.71, 173.85, 174.45
IV. Moc-Phe-Gly-Ala-Leu-H- 29.9 g (70.7 mmol) of Dess-Martin periodinane issuspended in 500 ml CH₂Cl₂ and stirred for 10 min. Moc-Phe-Gly-Ala-Leucinol(10.6 g, 23.5 mmol) iss dissolved in 100 ml CH₂Cl₂ and added at a moderate rate tothe periodinane slurry. The mixture is stirred for 1h and poured into 150 mlNaHCO₃ containing 123 g Na₂S₂O₃. The mixture is allowed to stir for 15 min andextracted with EtOAc. The organic phase is dried and filtered followed by removalof solvent. Chromatography (3.5% MeOH/CH₂Cl₂) on silica gives 5.1 g of purewhite solid that is a mixture of the methoxy hemiacetal and aldehyde.¹³C NMR(CDCl₃,CD₃OD) 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, 99.18,127.00, 128.60, 129.06, 136.44, 157.27, 169.19, 169.67, 172.73, 173.40, 200.43.
V. Moc-Phe-OH - L-Phenylalanine (5.0 g, 30.2 mmol) is dissolved in 30 ml 1NNaOH and cooled to 0°C. Methyl chloroformate (2.53 ml, 31.8 mmol) is addeddropwise while in a separate addition funnel 30 ml of 1N NaOH is addedsimultaneously. After addition is complete, the solution is washed with 200 mlEtOAc and the aqueous phase acidified to pH = 2. The mixture is extracted withEtOAc (2X 100 ml), dried (MgSO₄), filtered, and the solvent removed to afford 6.0g product¹³C NMR (CDCl₃) 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 Et₂Oat room temperature is added 0.36 ml (2.57 mmol) of TEA. This is followed by the 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 thephases separate. The organic phase is dried with MgSO₄, filtered and the solventremoved to give 0.66 g of product (96% yield).¹³C NMR (CDCl₃) 27.05, 38,47,53.45, 54.64, 65.90, 127.43, 127.85, 128.48, 129.28, 130.27, 135.23, 136.22, 158.17,173.08. To a solution of the crude product (2.11 mmol) in 25 ml MeOH is added0.120 g Pd/C and the slurry degassed. The slurry is stirred under a positive pressureof H₂ via balloon for 1.5 h. The slurry is filtered through Celite and the filter cakewashed with MeOH. The solvent is removed to afford 0.430 g product.¹³C NMR26.50, 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 (0.900mmol) and 0.253 g Gly-Ala-Leu-OMe.HCl (0.818 mmol, generated byhydrogenation of I., above, according to the procedure outlined for compound II.) in15 ml DMF is added 0.250 ml TEA (1.80 mmol) followed by the addition of 0.147ml 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 HCl,saturated NaHCO₃, and brine. The solution is dried, filtered and the solventremoved to give 0.300 g product. The crude product (0.628 mmol) is dissolved in17 ml EtOH and cooled to 0°C. To this solution is added 0.140 g CaCl₂ (1.25mmol) 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. Theorganic phase is dried with MgSO₄, filtered and the solvent removed.Chromatography with 4% MeOH/CH₂Cl₂ gave 0.200 g pure product.¹³C NMR(CD₃OD) 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 CH₂Cl₂ 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. Themixture is poured into saturated NaHCO₃ containing 2.32 g Na₂S₂O₃ and thesolution stirred for 10 min, followed by extraction with EtOAc. The organic phaseis dried with MgSO₄, filtered and the solvent removed. The residue ischromatographed on silica to give 0.081 g product.¹³C NMR (10% CD₃OD inCDCl₃) 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.
Cbz =
carbobenzyloxy
Gly =
glycine
Ala =
alanine
Leu =
leucine
Phe =
phenylalanine
OMe =
methyl ester
TEA =
triethylamine
DECP =
diethylcyanophosphonate
TLC =
thin layer chromatography
MeOH =
methanol
Pd/C =
palladium on activated carbon
EtOH =
ethanol
THF =
tetrahydrofuran
Mac =
methylaminocarbonyl
Moc =
methoxycarbonyl
Etoc =
ethoxycarbonyl
Ms =
methanesulfonyl
Proteolytic Enzyme - Another essential ingredient in the present liquiddetergent compositions is active proteolytic enzyme. Mixtures of proteolyticenzyme are also included. The proteolytic enzyme can be of animal, vegetable ormicroorganism (preferred) origin. The proteases for use in the detergentcompositions herein include (but are not limited to) trypsin, subtilisin, chymotrypsinand elastase-type proteases. Preferred for use herein are subtilisin-type proteolyticenzymes. Particularly preferred is bacterial serine proteolytic enzyme obtained fromBacillussubtilis and/orBacilluslicheniformis. Protease enzymes are usually presentin such liquid detergent compositions at levels sufficient to provide from 0.005 to0.1 Anson units (AU) of activity per gram of composition.
Suitable proteolytic enzymes include Novo Industri A/S Alcalase® (preferred),Esperase®, Savinase® (Copenhagen, Denmark), Gist-brocades' Maxatase®,Maxacal® and Maxapem 15® (protein engineered Maxacal®) (Delft, Netherlands),and subtilisin BPN and BPN'(preferred), which are commercially available.Preferred proteolytic enzymes are also modified bacterial serine proteases, such asthose made by Genencor International, Inc.(San Francisco, California) which aredescribed 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, Venegas, issued July 9, 1991, which refers to a modified bacterial serine proteolyticenzyme (Genencor International) which is called "Protease A" herein (same asBPN'). 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 mixturesthereof. Protease B is most preferred.
Another preferred protease, referred to as "Protease D" is a carbonylhydrolase variant having an amino acid sequence not found in nature, which isderived from a precursor carbonyl hydrolase by substituting a different amino acidfor a plurality of amino acid residues at a position in said carbonyl hydrolaseequivalent to position +76, preferably also in combination with one or more aminoacid residue positions equivalent to those selected from the group consisting of +99,+101, +103, +104, +107, +123, +27, +105, +109, +126, +128, +135, +156, +166,+195, +197, +204, +206, +210, +216, +217, +218, +222, +260, +265, and/or +274according to the numbering ofBacillus amyloliquefaciens subtilisin, as described inWO 95/10615 published April 20, 1995 by Genencor International.
Useful proteases are also described in PCT publications: WO 95/30010published Novenber 9, 1995 by The Procter & Gamble Company; WO 95/30011published Novenber 9, 1995 by The Procter & Gamble Company; WO 95/29979published Novenber 9, 1995 by The Procter & Gamble Company.
Calcium - Any water-soluble calcium salt can be used as a source of calciumions, including calcium acetate, calcium formate, calcium xylene sulfonate, andcalcium propionate. Divalent ions, such as zinc and magnesium ions, can replacethe calcium ion in part. Thus in the liquid detergent compositionsherein, the source of calcium ions can be partially substituted with a source ofanother divalent ion.
The calcium useful herein is enzyme-accessible. Therefore, the claimedcompositions are substantially free of sequestrants, for example, polyacids capableof forming calcium complexes which are soluble in the composition. However,minor amounts of sequestrants such as polyacids or mixtures of polyacids can beused. The enzyme-accessible calcium is defined as the amount of calcium-ionseffectively available to the enzyme component. From a practical standpoint theenzyme-accessible calcium is therefore the soluble calcium in the composition in theabsence of any storage sequestrants, e.g., having an equilibrium constant ofcomplexation with calcium equal to or greater than 1.5 at 20°C.
Boric Acid - The compositions herein optionally contain from about 0.25% toabout 10%, preferably from about 0.5% to about 5%, more preferably from about0.75% to about 3%, by weight of boric acid or a compound capable of forming boricacid in the composition (calculated on the basis of the boric acid). Boric acid ispreferred, although other compounds such as boric oxide, borax and other alkalimetal borates (e.g., sodium ortho-, meta-, pyroborate, an sodium pentaborate) aresuitable. Substituted boric acids (e.g., phenylboronic acid, butane boronic acid, andp-bromo phenylboronic acid) can also be used in place of boric acid.
The compositions of the present invention can also contain polyols, especiallydiols, containing only carbon, hydrogen and oxygen atoms. They preferably containfrom 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 about1% to about 15%, preferably from about 1.5% to about 10%, more preferably fromabout 2% to about 7%, by weight of the composition.
Detersive Surfactant - An effective amount, from 1 to 95,preferably 8 to 70, weight %, of detersive surfactant is yet another essentialingredient in the present invention. The detersive surfactant can be selected from thegroup consisting of anionics, nonionics, cationics, ampholytics, zwitterionics, andmixtures thereof. By selecting the type and amount of detersive surfactant, alongwith other adjunct ingredients disclosed herein, the present detergent compositionscan be formulated to be used in the context of laundry cleaning or in other differentcleaning applications, particularly including dishwashing. The particular surfactantsused can therefore vary widely depending upon the particular end-use envisioned.
The benefits of the present invention are especially pronounced incompositions containing ingredients that are harsh to enzymes such as certaindetergency builders and surfactants. These include (but are not limited to) anionicsurfactants such as alkyl ether sulfate linear alkyl benzene sulfonate, alkyl sulfate.Suitable surfactants are described below.
Anionic Surfactants - One type of anionic surfactant which can be utilizedencompasses alkyl ester sulfonates. These are desirable because they can be madewith renewable, non-petroleum resources. Preparation of the alkyl ester sulfonatesurfactant component can be effected according to known methods disclosed in thetechnical literature. For instance, linear esters of C₈-C₂₀ carboxylic acids can besulfonated with gaseous SO₃ according to "The Journal of the American OilChemists Society," 52 (1975), pp. 323-329. Suitable starting materials wouldinclude natural fatty substances as derived from tallow, palm, and coconut oils, etc.
The preferred alkyl ester sulfonate surfactant, especially for laundryapplications, comprises alkyl ester sulfonate surfactants of the structural formula:
wherein R³ is a C₈-C₂₀ hydrocarbyl, preferably an alkyl, or combination thereof,R⁴ is a C₁-C₆ hydrocarbyl, preferably an alkyl, or combination thereof, and M is asoluble salt-forming cation. Suitable salts include metal salts such as sodium,potassium, and lithium salts, and substituted or unsubstituted ammonium salts, suchas methyl-, dimethyl, -trimethyl, and quaternary ammonium cations, e.g.tetramethyl-ammonium and dimethyl piperdinium, and cations derived fromalkanolamines, e.g. monoethanol-amine, diethanolamine, and triethanolamine.Preferably, R³ is C₁₀-C₁₆ alkyl, and R⁴ is methyl, ethyl or isopropyl. Especiallypreferred are the methyl ester sulfonates wherein R³ is C₁₄-C₁₆ alkyl.
Alkyl sulfate surfactants are another type of anionic surfactant of importancefor use herein. In addition to providing excellent overall cleaning ability when usedin combination with polyhydroxy fatty acid amides (see below), including goodgrease/oil cleaning over a wide range of temperatures, wash concentrations, andwash times, dissolution of alkyl sulfates can be obtained, as well as improvedformulability in liquid detergent formulations are water soluble salts or acids of theformula ROSO₃M wherein R preferably is a C₁₀-C₂₄ hydrocarbyl, preferably analkyl or hydroxyalkyl having a C₁₀-C₂₀ alkyl component, more preferably a C₁₂-C₁₈alkyl or hydroxyalkyl, and M is H or a cation, e.g., an alkali metal cation (e.g.,sodium, potassium, lithium), substituted or unsubstituted ammonium cations such asmethyl-, dimethyl-, and trimethyl ammonium and quaternary ammonium cations,e.g., tetramethyl-ammonium and dimethyl piperdinium, and cations derived fromalkanolamines such as ethanolamine, diethanolamine, triethanolamine, and mixturesthereof, and the like. Typically, alkyl chains of C_12-16 are preferred for lower washtemperatures (e.g., below about 50°C) and C_16-18 alkyl chains are preferred forhigher wash temperatures (e.g., above about 50°C).
Alkyl alkoxylated sulfate surfactants are another category of useful anionicsurfactant. These surfactants are water soluble salts or acids typically of the formulaRO(A)_mSO₃M wherein R is an unsubstituted C₁₀-C₂₄ alkyl or hydroxyalkyl grouphaving a C₁₀-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 preferablybetween about 0.5 and about 3, and M is H or a cation which can be, for example, a metal cation (e.g., sodium, potassium, lithium, calcium, magnesium, etc.),ammonium or substituted-ammonium cation. Alkyl ethoxylated sulfates as well asalkyl propoxylated sulfates are contemplated herein. Specific examples ofsubstituted ammonium cations include methyl-, dimethyl-, trimethyl-ammonium andquaternary ammonium cations, such as tetramethyl-ammonium, dimethylpiperidinium and cations derived from alkanolamines, e.g. monoethanolamine,diethanolamine, and triethanolamine, and mixtures thereof. Exemplary surfactantsare C₁₂-C₁₈ alkyl polyethoxylate (1.0) sulfate, C₁₂-C₁₈ alkyl polyethoxylate (2.25)sulfate, C₁₂-C₁₈ alkyl polyethoxylate (3.0) sulfate, and C₁₂-C₁₈ alkylpolyethoxylate (4.0) sulfate wherein M is conveniently selected from sodium andpotassium.
Other Anionic Surfactants - Other anionic surfactants useful for detersivepurposes can also be included in the compositions hereof. These can include salts(including, for example, sodium, potassium, ammonium, and substituted ammoniumsalts such as mono-, di- and triethanolamine salts) of soap, C₉-C₂₀ linearalkylbenzenesulphonates, C₈-C₂₂ primary or secondary alkanesulphonates, C₈-C₂₄olefinsulphonates, sulphonated polycarboxylic acids prepared by sulphonation of thepyrolyzed product of alkaline earth metal citrates, e.g., as described in British patentspecification No. 1,082,179, alkyl glycerol sulfonates, fatty acyl glycerol sulfonates,fatty oleyl glycerol sulfates, alkyl phenol ethylene oxide ether sulfates, paraffinsulfonates, alkyl phosphates, isothionates such as the acyl isothionates, N-acyltaurates, fatty acid amides of methyl tauride, alkyl succinamates and sulfosuccinates,monoesters of sulfosuccinate (especially saturated and unsaturated C₁₂-C₁₈monoesters) diesters of sulfosuccinate (especially saturated and unsaturated C₆-C₁₄diesters), N-acyl sarcosinates, sulfates of alkylpolysaccharides such as the sulfates ofalkylpolyglucoside (the nonionic nonsulfated compounds being described below),branched primary alkyl sulfates, alkyl polyethoxy carboxylates such as those of theformula RO(CH₂CH₂O)_kCH₂COO-M⁺ wherein R is a C₈-C₂₂ alkyl, k is aninteger from 0 to 10, and M is a soluble salt-forming cation, and fatty acidsesterified with isethionic acid and neutralized with sodium hydroxide. Resin acidsand hydrogenated 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 examples are given in "Surface Active Agents and Detergents" (Vol. I and IIby Schwartz, Perry and Berch). A variety of such surfactants are also generallydisclosed in U.S. Patent 3,929,678, issued December 30, 1975 to Laughlin, et al. atColumn 23, line 58 through Column 29, line 23.
Nonionic Detergent Surfactants - Suitable nonionic detergent surfactants aregenerally disclosed in U.S. Patent 3,929,678, Laughlin et al., issued December 30,1975, at column 13, line 14 through column 16, line 6.Exemplary, non-limiting classes of useful nonionic surfactants are listedbelow.
The polyethylene, polypropylene, and polybutylene oxide condensates of alkylphenols. In general, the polyethylene oxide condensates are preferred. Thesecompounds include the condensation products of alkyl phenols having an alkylgroup containing from about 6 to about 12 carbon atoms in either a straight chain orbranched 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 molesof ethylene oxide per mole of alkyl phenol. Commercially available nonionicsurfactants of this type include Igepal® CO-630, marketed by the GAF Corporation;and Triton® X-45, X-114, X-100, and X-102, all marketed by the Rohm & HaasCompany. These compounds are commonly referred to as alkyl phenol alkoxylates,(e.g., alkyl phenol ethoxylates).
The condensation products of aliphatic alcohols with from about 1 to about 25moles of ethylene oxide. The alkyl chain of the aliphatic alcohol can either bestraight or branched, primary or secondary, and generally contains from about 8 toabout 22 carbon atoms. Particularly preferred are the condensation products ofalcohols having an alkyl group containing from about 10 to about 20 carbon atomswith from about 2 to about 18 moles of ethylene oxide per mole of alcohol.Examples of commercially available nonionic surfactants of this type includeTergitol® 15-S-9 (the condensation product of C₁₁-C₁₅ linear secondary alcoholwith 9 moles ethylene oxide), Tergitol® 24-L-6 NMW (the condensation product ofC₁₂-C₁₄ primary alcohol with 6 moles ethylene oxide with a narrow molecularweight distribution), both marketed by Union Carbide Corporation; Neodol® 45-9(the condensation product of C₁₄-C₁₅ linear alcohol with 9 moles of ethyleneoxide), Neodol® 23-6.5 (the condensation product of C₁₂-C₁₃ linear alcohol with6.5 moles of ethylene oxide), Neodol® 45-7 (the condensation product of C₁₄-C₁₅linear alcohol with 7 moles of ethylene oxide), Neodol® 45-4 (the condensationproduct of C₁₄-C₁₅ linear alcohol with 4 moles of ethylene oxide), marketed byShell Chemical Company, and Kyro® EOB (the condensation product of C₁₃-C₁₅alcohol with 9 moles ethylene oxide), marketed by The Procter & Gamble Company.This category of nonionic surfactant is referred to generally as "alkyl ethoxylates."
The condensation products of ethylene oxide with a hydrophobic base formedby the condensation of propylene oxide with propylene glycol. The hydrophobic portion of these compounds preferably has a molecular weight of from about 1500 toabout 1800 and exhibits water insolubility. The addition of polyoxyethylenemoieties to this hydrophobic portion tends to increase the water solubility of themolecule as a whole, and the liquid character of the product is retained up to thepoint where the polyoxyethylene content is about 50% of the total weight of thecondensation product, which corresponds to condensation with up to about 40 molesof ethylene oxide. Examples of compounds of this type include certain of thecommercially-available Pluronic® surfactants, marketed by BASF.
The condensation products of ethylene oxide with the product resulting fromthe reaction of propylene oxide and ethylenediamine. The hydrophobic moiety ofthese products consists of the reaction product of ethylenediamine and excesspropylene oxide, and generally has a molecular weight of from about 2500 to about3000. This hydrophobic moiety is condensed with ethylene oxide to the extent thatthe condensation product contains from about 40% to about 80% by weight ofpolyoxyethylene 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 commerciallyavailable Tetronic® compounds, marketed by BASF.
Semi-polar nonionic surfactants are a special category of nonionic surfactantswhich include water-soluble amine oxides containing one alkyl moiety of fromabout 10 to about 18 carbon atoms and 2 moieties selected from the group consistingof alkyl groups and hydroxyalkyl groups containing from about 1 to about 3 carbonatoms; water-soluble phosphine oxides containing one alkyl moiety of from about 10to about 18 carbon atoms and 2 moieties selected from the group consisting of alkylgroups and hydroxyalkyl groups containing from about 1 to about 3 carbon atoms;and water-soluble sulfoxides containing one alkyl moiety of from about 10 to about18 carbon atoms and a moiety selected from the group consisting of alkyl andhydroxyalkyl moieties of from about 1 to about 3 carbon atoms.
Semi-polar nonionic detergent surfactants include the amine oxide surfactantshaving the formula
wherein R³ is an alkyl, hydroxyalkyl, or alkyl phenyl group or mixtures thereofcontaining from about 8 to about 22 carbon atoms; R⁴ is an alkylene orhydroxyalkylene group containing from about 2 to about 3 carbon atoms or mixturesthereof; x is from 0 to about 3; and each R⁵ is an alkyl or hydroxyalkyl groupcontaining from about 1 to about 3 carbon atoms or a polyethylene oxide groupcontaining from about 1 to about 3 ethylene oxide groups. The R⁵ groups can be attached to each other, e.g., through an oxygen or nitrogen atom, to form a ringstructure.
These amine oxide surfactants in particular include C₁₀-C₁₈ alkyl dimethylamine oxides and C₈-C₁₂ alkoxy ethyl dihydroxy ethyl amine oxides.
Alkylpolysaccharides disclosed in U.S. Patent 4,565,647, Llenado, issuedJanuary 21, 1986, having a hydrophobic group containing from about 6 to about 30carbon atoms, preferably from about 10 to about 16 carbon atoms and apolysaccharide, e.g., a polyglycoside, hydrophilic group containing from about 1.3to about 10, preferably from about 1.3 to about 3, most preferably from about 1.3 toabout 2.7 saccharide units. Any reducing saccharide containing 5 or 6 carbon atomscan be used, e.g., glucose, galactose and galactosyl moieties can be substituted forthe 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 orgalactoside.) The intersaccharide bonds can be, e.g., between the one position of theadditional saccharide units and the 2-, 3-, 4-, and/or 6- positions on the precedingsaccharide units.
Optionally, and less desirably, there can be a polyalkylene-oxide chain joiningthe hydrophobic moiety and the polysaccharide moiety. The preferred alkyleneoxideis ethylene oxide. Typical hydrophobic groups include alkyl groups, either saturatedor unsaturated, branched or unbranched containing from about 8 to about 18,preferably from about 10 to about 16, carbon atoms. Preferably, the alkyl group is astraight chain saturated alkyl group. The alkyl group can contain up to about 3hydroxy groups and/or the polyalkyleneoxide chain can contain up to about 10,preferably less than 5, alkyleneoxide moieties. Suitable alkyl polysaccharides areoctyl, nonyl, decyl, undecyldodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl,heptadecyl, and octadecyl, di-, tri-, tetra-, penta-, and hexaglucosides, galactosides,lactosides, glucoses, fructosides, fructoses and/or galactoses. Suitable mixturesinclude coconut alkyl, di-, tri-, tetra-, and pentaglucosides and tallow alkyl tetra-,penta-, and hexa-glucosides.
The preferred alkylpolyglycosides have the formulaR²O(C_nH_2nO)_t(glycosyl)_xwherein 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, carbonatoms; n is 2 or 3, preferably 2; t is from 0 to about 10, preferably 0; and x is fromabout 1.3 to about 10, preferably from about 1.3 to about 3, most preferably fromabout 1.3 to about 2.7. The glycosyl is preferably derived from glucose. To prepare these compounds, the alcohol or alkylpolyethoxy alcohol is formed first and thenreacted with glucose, or a source of glucose, to form the glucoside (attachment at the1-position). The additional glycosyl units can then be attached between their 1-positionand the preceding glycosyl units 2-, 3-, 4- and/or 6-position, preferablypredominantly the 2-position.
Fatty acid amide surfactants having the formula:
wherein R⁶ is an alkyl group containing from about 7 to about 21 (preferably fromabout 9 to about 17) carbon atoms and each R⁷ is selected from the group consistingof hydrogen, C₁-C₄ alkyl, C₁-C₄ hydroxyalkyl, and -(C²H₄O)_xH where x variesfrom about 1 to about 3.
Preferred amides are C₈-C₂₀ ammonia amides, monoethanolamides,diethanolamides, and isopropanolamides.
Cationic Surfactants - Cationic detersive surfactants can also be included indetergent compositions of the present invention. Cationic surfactants include theammonium surfactants such as alkyldimethylammonium halogenides, and thosesurfactants 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 carbonatoms in the alkyl chain, each R³ is selected from the group consisting of -CH₂CH₂-,-CH₂CH(CH₃)-, -CH₂CH(CH₂OH)-, -CH₂CH₂CH₂-, and mixtures thereof; eachR⁴ is selected from the group consisting of C₁-C₄ alkyl, C₁-C₄ hydroxyalkyl,benzyl, ring structures formed by joining the two R⁴ groups,-CH₂CHOHCHOHCOR⁶CHOH-CH₂OH wherein R⁶ is any hexose or hexosepolymer having a molecular weight less than about 1000, and hydrogen when y isnot O; R⁵ is the same as R⁴ or is an alkyl chain wherein the total number of carbonatoms of R² plus R⁵ is not more than about 18; each y is from 0 to about 10 and thesum of the y values is from 0 to about 15; and X is any compatible anion.
Other cationic surfactants useful herein are also described in U.S. Patent4,228,044, Cambre, issued October 14,1980,
Other Surfactants - Ampholytic surfactants can be incorporated into thedetergent compositions hereof. These surfactants can be broadly described asaliphatic derivatives of secondary or tertiary amines, or aliphatic derivatives ofheterocyclic secondary and tertiary amines in which the aliphatic radical can bestraight chain or branched. One of the aliphatic substituents contains at least about 8carbon atoms, typically from about 8 to about 18 carbon atoms, and at least one contains an anionic water-solubilizing group, e.g., carboxy, sulfonate, sulfate. SeeU.S. Patent No. 3,929,678 to Laughlin et al., issued December 30, 1975 at column19, lines 18-35 for examples of ampholytic surfactants.
Zwitterionic surfactants can also be incorporated into the detergentcompositions hereof. These surfactants can be broadly described as derivatives ofsecondary and tertiary amines, derivatives of heterocyclic secondary and tertiaryamines, or derivatives of quaternary ammonium, quaternary phosphonium or tertiarysulfonium compounds. See U.S. Patent No. 3,929,678 to Laughlin et al., issuedDecember 30, 1975 at column 19, line 38 through column 22, line 48 for examplesof zwitterionic surfactants. Ampholytic and zwitterionic surfactants are generallyused in combination with one or more anionic and/or nonionic surfactants.
Polyhydroxy Fatty Acid Amide Surfactant - The liquid detergent compositionshereof may also contain an enzyme-enhancing amount of polyhydroxy fatty acidamide surfactant. By "enzyme-enhancing" is meant that the formulator of thecomposition can select an amount of polyhydroxy fatty acid amide to beincorporated 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% weightbasis, polyhydroxy fatty acid amide surfactant, preferably from about 3% to about30%, of the polyhydroxy fatty acid amide. The polyhydroxy fatty acid amidesurfactant component comprises compounds of the structural formula:
wherein: R¹ is H, C₁-C₄ hydrocarbyl, 2-hydroxy ethyl, 2-hydroxy propyl, or amixture thereof, preferably C₁-C₄ alkyl, more preferably C₁ or C₂ alkyl, mostpreferably C₁ alkyl (i.e., methyl); and R² is a C₅-C₃₁ hydrocarbyl, preferablystraight chain C₇-C₁₉ alkyl or alkenyl, more preferably straight chain C₉-C₁₇ alkylor alkenyl, most preferably straight chain C₁₁-C₁₅ alkyl or alkenyl, or mixturesthereof; and Z is a polyhydroxyhydrocarbyl having a linear hydrocarbyl chain withat least 3 hydroxyls directly connected to the chain, or an alkoxylated derivative(preferably ethoxylated or propoxylated) thereof. Z preferably will be derived froma reducing sugar in a reductive amination reaction; more preferably Z will be aglycityl. 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 individual sugars listed above. These corn syrups may yield a mix of sugarcomponents for Z. It should be understood that it is by no means intended toexclude other suitable raw materials. Z preferably will be selected from the groupconsisting of -CH₂-(CHOH)_n-CH₂OH, -CH(CH₂OH)-(CHOH)_n-1-CH₂OH, -CH₂-(CHOH)₂(CHOR')(CHOH)-CH₂OH,and alkoxylated derivatives thereof, where n isan 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 example, N-methyl, N-ethyl, N-propyl, N-isopropyl, N-butyl, N-2-hydroxyethyl, or N-2-hydroxy propyl.
R²-CO-N< can be, for example, cocamide, stearamide, oleamide, lauramide,myristamide, capricamide, palmitamide, tallowamide, etc.
Z can be 1-deoxyglucityl, 2-deoxyfructityl, 1-deoxymaltityl, 1-deoxylactityl, 1-deoxygalactityl,1-deoxymannityl, 1-deoxymaltotriotityl, etc.
Methods for making polyhydroxy fatty acid amides are known in the art. Ingeneral, they can be made by reacting an alkyl amine with a reducing sugar in areductive amination reaction to form a corresponding N-alkyl polyhydroxyamine,and then reacting the N-alkyl polyhydroxyamine with a fatty aliphatic ester ortriglyceride in a condensation/amidation step to form the N-alkyl, N-polyhydroxyfatty acid amide product. Processes for making compositions containingpolyhydroxy fatty acid amides are disclosed, for example, in G.B. PatentSpecification 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.
Second Enzyme - Preferred compositions herein further comprise aperformance-enhancing amount of a detergent-compatible second enzyme. By"detergent-compatible" is meant compatibility with the other ingredients of a liquiddetergent composition, such as detersive surfactant and detergency builder. Thesesecond enzymes are preferably selected from the group consisting of lipase, amylase,cellulase, and mixtures thereof. The term "second enzyme" excludes the proteolyticenzymes discussed above, so each composition which has a second enzyme containsat least two kinds of enzyme, including at least one proteolytic enzyme. The amountof 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 % ofthese second enzymes are preferably used. Mixtures of the same class of enzymes (e.g. lipase) or two or more classes (e.g. cellulase and lipase) may be used. Purifiedor non-purified forms of the enzyme may be used.
Any lipolytic enzyme suitable for use in a liquid detergent composition can beused in these compositions. Suitable lipase enzymes for use herein include those ofbacterial and fungal origin.
Suitable bacterial lipases include those produced by microorganisms of thePseudomonas groups, such asPseudomonasstutzeri ATCC 19.154, as disclosed inBritish Patent 1,372,034. Suitable lipases includethose which show a positive immunological cross-reaction with the antibody of thelipase produced by the microorganismPseudomonasfluorescens IAM 1057. Thislipase and a method for its purification have been described in Japanese PatentApplication 53-20487, laid open on February 24, 1978. This lipase is available fromAmano Pharmaceutical Co. Ltd., Nagoya, Japan, under the trade name Lipase P"Amano," hereinafter referred to as "Amano-P." Such lipases should show apositive immunological cross-reaction with the Amano-P antibody, using thestandard and well-known immunodiffusion procedure according to Ouchterlony(Acta. Med. Scan., 133, pages 76-79 (1950)). These lipases, and a method for theirimmunological cross-reaction with Amano-P, are also described in U.S. Patent4,707,291, Thom et al., issued November 17, 1987.Typical examples thereof are the Amano-P lipase, the lipase exPseudomonasfragiFERM P 1339 (available under the trade name Amano-B), lipase exPseudomonasnitroreducens var.lipolyticum FERM P 1338 (available under the trade nameAmano-CES), lipases exChromobacterviscosum, e.g.Chromobacterviscosum var.lipolyticum NRRLB 3673, commercially available from Toyo Jozo Co., Tagata,Japan; and furtherChromobacterviscosum lipases from U.S. Biochemical Corp.,U.S.A. and Disoynth Co., The Netherlands, and lipases exPseudomonasgladioli.
Suitable fungal lipases include those producible byHumicolalanuginosa andThermomyceslanuginosus. Most preferred is lipase obtained by cloning the genefromHumicolalanuginosa and expressing the gene inAspergillusoryzae asdescribed 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 gram(LU/g) of lipase can be used in these compositions. A lipase unit is that amount oflipase which produces 1 mmol of titratable fatty acid per minute in a pH stat, wherepH is 9.0, temperature is 30°C, substrate is an emulsion of 3.3wt % of olive oil and3.3% gum arabic, in the presence of 13 mmol/l Ca⁺⁺ and 20 mmol/l NaCl in 5mmol/l Tris-buffer.
Any cellulase suitable for use in a liquid detergent composition can be used inthese compositions. Suitable cellulase enzymes for use herein include those frombacterial and fungal origins. Preferably, they will have a pH optimum of between 5and 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 fungalcellulase produced fromHumicolainsolens. Suitable cellulases are also disclosed inGB-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 ofHumicolainsolens(Humicolagrisea var.thermoidea), particularly the Humicola strain DSM1800, and cellulases produced by a fungus ofBacillus N or a cellulase 212-producingfungus belonging to the genusAeromonas, and cellulase extracted fromthe hepatopancreas of a marine mollusc (Dolabella Auricula Solander).
Any amylase suitable for use in a liquid detergent composition can be used inthese compositions. Amylases include, for example, amylases obtained from aspecial strain ofB.licheniformis, described in more detail in British PatentSpecification No. 1,296,839 (Novo). Amylolytic proteins include, for example,Rapidase^R, International Bio-Synthetics, Inc. and Termamyl^R Novo Industries.
From about 0.0001% to 0.55, preferably 0.0005 to 0.1, wt. % amylase can beused.
Optional Ingredients - Detergent builders can optionally be included in thecompositions herein, especially for laundry compositions. Inorganic as well asorganic builders can be used. When present, the compositions will typicallycomprise at least about 1% builder and can be either an inorganic or organic builder.Liquid laundry formulations preferably comprise from about 3% to 30%, morepreferably about 5 to 20%, by weight, of detergent builder.
Inorganic detergent builders include, but are not limited to, the alkali metal,ammonium and alkanolammonium salts of polyphosphates (exemplified by thetripolyphosphates, pyrophosphates, and glassy polymeric meta-phosphates),phosphonates, phytic acid, silicates, carbonates (including bicarbonates andsesquicarbonates), sulphates, and aluminosilicates. Borate builders, as well asbuilders containing borate-forming materials that can produce borate under detergentstorage or wash conditions (hereinafter, collectively "borate builders"), can also beused. Preferably, non-borate builders are used in the compositions of the inventionintended for use at wash conditions less than about 50°C, especially less than about40°C.
Examples of silicate builders are the alkali metal silicates, particularly thosehaving a SiO₂:Na₂O ratio in the range 1.6:1 to 3.2:1 and layered silicates, such asthe layered sodium silicates described in U.S. Patent 4,664,839, issued May 12,1987 to H. P. Rieck. However, other silicates mayalso be useful such as for example magnesium silicate, which can serve as acrispening 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 alkaline earth and alkali metalcarbonates, including sodium carbonate and sesquicarbonate and mixtures thereofwith ultra-fine calcium carbonate as disclosed in German Patent Application No.2,321,001 published on November 15, 1973.
Aluminosilicate builders are useful in the present invention. Aluminosilicatebuilders are of great importance in most currently marketed heavy duty granulardetergent compositions, and can also be a significant builder ingredient in liquiddetergent formulations. Aluminosilicate builders include those having the empiricalformula:M_z(zAlO_2·ySiO₂)wherein M is sodium, potassium, ammonium or substituted ammonium, z is fromabout 0.5 to about 2; and y is 1; this material having a magnesium ion exchangecapacity of at least about 50 milligram equivalents of CaCO₃ hardness per gram ofanhydrous aluminosilicate. Preferred alumino-silicates are zeolite builders whichhave the formula:Na_z[(AlO₂)_z(SiO₂)_y].xH₂Owherein z and y are integers of at least 6, the molar ratio of z to y is in the rangefrom 1.0 to about 0.5, and x is an integer from about 15 to about 264.
Useful aluminosilicate ion exchange materials are commercially available.These aluminosilicates can be crystalline or amorphous in structure and can benaturally-occurring aluminosilicates or synthetically derived. A method forproducing aluminosilicate ion exchange materials is disclosed in U.S. Patent3,985,669, Krummel, et al., issued October 12, 1976.Preferred synthetic crystalline aluminosilicate ion exchange materialsuseful herein are available under the designations Zeolite A, Zeolite P (B), andZeolite X. In an especially preferred embodiment, the crystalline aluminosilicate ionexchange material has the formula:Na₁₂[(AlO₂)₁₂(SiO₂)₁₂]^·xH₂O wherein x is from about 20 to about 30, especially about 27. This material is knownas Zeolite A. Preferably, the aluminosilicate has a particle size of about 0.1-10microns in diameter.
Specific examples of polyphosphates are the alkali metal tripolyphosphates,sodium, potassium and ammonium pyrophosphate, sodium and potassium andammonium pyrophosphate, sodium and potassium orthophosphate, sodium polymetaphosphate 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 -hydroxy-1, 1-diphosphonate particularly the sodium and potassium salts, the water-solublesalts of methylene diphosphonic acid e.g. the trisodium and tripotassiumsalts and the water-soluble salts of substituted methylene diphosphonic acids, suchas the trisodium and tripotassium ethylidene, isopyropylidene benzylmethylideneand halo methylidene phosphonates. Phosphonate builder salts of theaforementioned types are disclosed in U.S. Patent Nos. 3,159,581 and 3,213,030issued December 1, 1964 and October 19, 1965, to Diehl; U.S. Patent No. 3,422,021issued January 14, 1969, to Roy; and U.S. Patent Nos. 3,400,148 and 3,422,137issued September 3, 1968, and January 14, 1969 to Quimby.
Organic detergent builders preferred for the purposes of the present inventioninclude 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 acidform, but can also be added in the form of a neutralized salt. When utilized in saltform, alkali metals, such as sodium, potassium, and lithium, or alkanolammoniumsalts are preferred.
Included among the polycarboxylate builders are a variety of categories ofuseful materials. One important category of polycarboxylate builders encompassesthe ether polycarboxylates. A number of ether polycarboxylates have been disclosedfor 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.
A specific type of ether polycarboxylates useful as builders in the presentinvention also include those having the general formula:CH(A)(COOX)-CH(COOX)-O-CH(COOX)-CH(COOX)(B) wherein A is H or OH; B is H or -O-CH(COOX)-CH₂(COOX); and X is H or a salt-formingcation. For example, 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 andB is H, then the compound is tartrate monosuccinic acid (TMS) and its water-solublesalts. If A is H and B is -O-CH(COOX)-CH₂(COOX), then the compound is tartratedisuccinic acid (TDS) and its water-soluble salts. Mixtures of these builders areespecially preferred for use herein. Particularly preferred are mixtures of TMS andTDS in a weight ratio of TMS to TDS of from about 97:3 to about 20:80. Thesebuilders are disclosed in U.S. Patent 4,663,071, issued to Bush et al., on May 5,1987.
Suitable ether polycarboxylates also include cyclic compounds, particularlyalicyclic 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.
Other useful detergency builders include the ether hydroxypolycarboxylatesrepresented by the structure:HO-[C(R)(COOM)-C(R)(COOM)-O]_n-Hwherein M is hydrogen or a cation wherein the resultant salt is water-soluble,preferably an alkali metal, ammonium or substituted ammonium cation, n is fromabout 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 selectedfrom hydrogen, C_1-4 alkyl or C_1-4 substituted alkyl (preferably R is hydrogen).
Still other ether polycarboxylates include copolymers of maleic anhydride withethylene 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,ammonium and substituted ammonium salts of polyacetic acids. Examples includethe sodium, potassium, lithium, ammonium and substituted ammonium salts ofethylenediamine 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, andcarboxymethyloxysuccinic acid, and soluble salts thereof.
Citrate builders, e.g., citric acid and soluble salts thereof (particularly sodiumsalt), are polycarboxylate builders of particular importance for heavy duty liquiddetergent formulations, but can also be used in granular compositions.
Other carboxylate builders include the carboxylated carbohydrates disclosed inU.S. Patent 3,723,322, Diehl, issued March 28, 1973.
Also suitable in the detergent compositions of the present invention are the 3,3-dicarboxy-4-oxa-1,6-hexanedioatesand the related compounds disclosed in U.S.Patent 4,566,984, Bush, issued January 28, 1986.Useful succinic acid builders include the C₅-C₂₀ alkyl succinic acids and saltsthereof. A particularly preferred compound of this type is dodecenylsuccinic acid.Alkyl succinic acids typically are of the general formulaR-CH(COOH)CH₂(COOH) i.e., derivatives of succinic acid, wherein R ishydrocarbon, e.g., C₁₀-C₂₀ alkyl or alkenyl, preferably C₁₂-C₁₆ or wherein R maybe substituted with hydroxyl, sulfo, sulfoxy or sulfone substituents, all as describedin the above-mentioned patents.
The succinate builders are preferably used in the form of their water-solublesalts, including the sodium, potassium, ammonium and alkanolammonium salts.
Specific examples of succinate builders include: laurylsuccinate,myristylsuccinate, palmitylsuccinate, 2-dodecenylsuccinate (preferred), 2-pentadecenylsuccinate,and the like. Laurylsuccinates are the preferred builders ofthis group, and are described in European Patent Application 0,200,263,published November 5, 1986.
Examples of useful builders also include sodium and potassiumcarboxymethyloxymalonate, carboxymethyloxysuccinate, cis-cyclo-hexane-hexacarboxylate,cis-cyclopentane-tetracarboxylate, water-soluble polyacrylates(these polyacrylates having molecular weights to above about 2,000 can also beeffectively utilized as dispersants), and the copolymers of maleic anhydride withvinyl methyl ether or ethylene.
Other suitable polycarboxylates are the polyacetal carboxylates disclosed inU.S. Patent 4,144,226, Crutchfield et al., issued March 13, 1979.These polyacetal carboxylates can be prepared by bringing together,under polymerization conditions, an ester of glyoxylic acid and a polymerizationinitiator. The resulting polyacetal carboxylate ester is then attached to chemicallystable end groups to stabilize the polyacetal carboxylate against rapiddepolymerization in alkaline solution, converted to the corresponding salt, and addedto a surfactant.
Polycarboxylate builders are also disclosed in U.S. Patent 3,308,067, Diehl,issued March 7, 1967. Such materials include the water-soluble salts of homo- and copolymers of aliphatic carboxylic acids such asmaleic 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 canbe utilized. These would include materials generally referred to as "soaps." Chainlengths of C₁₀-C₂₀ are typically utilized. The hydrocarbyls can be saturated orunsaturated.
Other optional ingredients include soil release agents, chelating agents, claysoil removal/anti redeposition agents, polymeric dispersing agents, bleaches,brighteners, suds suppresors, solvents and aesthetic agents.
The detergent composition herein can be formulated as a variety ofcompositions, for instance as laundry detergents as well as hard surface cleaners ordishwashing compositions.
The compositions according to the present invention are further illustrated bythe following examples.

EXAMPLE I

The following compositions are made by combining the listed ingredients inthe listed proportions. In this example, one or more of the following peptidealdehydes are used:
Peptide aldehyde 1: 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
Peptide aldehyde 5: CH₃SO₂Phe-Gly-Ala-Leu-H
Peptide aldehyde 6: CH₃SO₂Val-Ala-Leu-H
Peptide aldehyde 7: C₆H₅CH₂O(OH)(O)P-Val-Ala-Leu-H
Peptide aldehyde 8: CH₃CH₂SO₂-Phe-Gly-Ala-Leu-H
Peptide aldehyde 9: C₆H₅CH₂SO₂-Val-Ala-Leu-H
Peptide aldehyde 10: C₆H₅CH₂O(OH)(O)P-Leu-Ala-Leu-H
Peptide aldehyde 11: C₆H₅CH₂O(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
Linear alkyl benzene sulfonic acid	8.5	15	6.5	10	12.5	4
Sodium C_12-15 alkyl sulfate	1	2	1	2	--	--
C_14-15 alkyl 2.5 times ethoxylated sulfate	10	5	10.5	--	11	9
C₁₂ glucose amide	--	--	9	--	--	5
C_12-15 alcohol 7 times ethoxylated	3	10	4	7	2.5	--
Fatty acid	2	5	5	4	2	2
Citric acid	6	7	4	6	4	5
C_12-14 alkenyl substituted, succinic acid	--	6	--	5	--	6
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® (100KLU/g commercial solution)	0.5	0.2	0.5	0.5	0.4	--
Protease B (34 g/L commerical solution)	0.9	--	0.5	--	1.2	--
Savinase® (commercial solution)	--	0.3	--	0.4	0.2	0.3
Carezyme®	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 formula is tested for % of protease activity remaining.Combinations of 0%, 0.1%, 0.2%, and 0.3% Ca⁺⁺ (from CaCl₂) and 0%, 0.0006%,0.00125%, and 0.0025% peptide aldehyde (Synthesis Example 6) are used. Productsare held at 32,2°C (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 (C11E9)	5
Mg ion from MgCl₂	1
Ca ion from CaCl₂	see chart below
Peptide aldehyde	see chart below
Sodium xylene sulfonate	4
Solvent	6
Water	to 100%
pH	to 8

EXAMPLE III

Results showing the percent protease activity remaining after 42 days at 32,2°C (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 inthe listed proportions.

Ingredients	A(wt%)	B(wt%)	C(wt%)	D(wt%)
LAS	0	0	0	12
AExS	22.1	24.7	33.5	3
Polyhydroxy fatty acid amide	4.6	1.2	4.2	0
Amine Oxide	4.6	1.2	4.8	0
Betaine	0	1.2	0	0
Nonionic Surfactant	6.7	4.1	0	0
Mg(OH)₂	0.5	0.5	0.7	0
Ca ion from CaCl₂	0.1	0.3	0.4	0.1
Calcium xylene sulfonate	4.5	0	4	0
Polyethylene glycol	3	0	0	0
Polypropylene glycol 2000	1.5	0	0	0
Balance, water	to 100%	to 100%	to 100%	to 100%
Protease A or Protease B	0.001-0.01	0.001-0.01	0.005-0.01	0.0003-0.01
Peptide Aldehydes	0.00025-0.0025	0.00025-0.0025	0.00025-0.0025	0.00125-0.0025

Claims

A liquid detergent composition comprising:
a) from 1% to 95%, by weight of composition of a detersive surfactant:
b) an active proteolytic enzyme;
c) a source of calcium ions other than a compound of the formulaRO(A)_mSO₃M wherein R is an unsubstituted C₁₀ - C₂₄ alkyl or hydroxyalkylgroup having a C₁₀ - C₂₄ alkyl component, A is an ethoxy or propoxy unit, mis greater that zero and M is calcium; 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 anN-capping moiety selected from the group consisting of phosphoramidate[(R"O)₂(O)P-], 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 groupconsisting of straight or branched C₁-C₆ unsubstituted alkyl, phenyl, C₇-C₉alkylaryl, and cycloalkyl moieties, wherein the cycloalkyl ring may span C₄-C₈and may contain one or more heteroatoms selected from the group consisting ofO,N,and S; and R is selected from the group consisting of straight or branchedC₁ - C₆ unsubstituted alkyl, phenyl, and C₇-C₉ alkylaryl moieties.
A liquid detergent composition according to Claim 1 wherein the source ofcalcium ion is selected from calcium formate, calcium chloride, calcium acetate,calcium xylene sulfonate, calcium sulfate, and mixtures thereof.
A liquid detergent composition comprising:
a) from 1% to 95%, by weight of composition, of a detersive surfactant;
b) an active proteolytic enzyme;
c) a source ofcalcium ion selected from calcium formate, calcium chloride, calcium acetate,calcium xylene sulfonate, calcium sulfate, and mixtures thereof;
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 anN-capping moiety selected from the group consisting of phosphoramidate[(R"O)₂(O)P-], 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 groupconsisting of straight or branched C₁-C₆ unsubstituted alkyl, phenyl, C₇-C₉alkylaryl, and cycloalkyl moieties, wherein the cycloalkyl ring may span C₄-C₈and may contain one or more heteroatoms selected from the group consisting ofO,N,and S; and R is selected from the group consisting of straight or branchedC₁ - C₆ unsubstituted alkyl, phenyl, and C₇ - C₉ alkylaryl moieties.
A liquid detergent composition according to Claim 1 or 3 comprising:
a) from 8 to 70% of said detersive surfactant;
b) from 0.0001% to 5% of an active proteolytic enzyme;
c) from 0.01% to 1% of calcium ion; and
d) from 0.00001% to 5% of said peptide aldehyde.
A liquid detergent composition according to Claim 1 or 3 wherein said R moieties areselected from the group consisting of methyl, iso-propyl, sec-butyl, iso-butyl,-C₆H₅,-CH₂-C₆H₅, and -CH₂CH₂-C₆H₅.
A liquid detergent composition according to Claim 5 wherein said B peptidechains are selected from the group consisting of peptide chains having the aminoacid sequences according to the general formula:Z-A⁵-A⁴-A³-A²-A¹-NH-CH(R)-C(O)Hsuch that the following amino acids, when present, are :
A¹ 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;
A⁵, if present, is any amino acid.
A liquid detergent composition according to Claim 6 wherein the N-terminal endis protected by one of the N-capping moiety protecting groups selected from thegroup consisting of carbamates, ureas, sulfonamides, phosphonamides, thioureas,sulfonamides, sulfonic acids, phosphinamides, thiocarbamates, amidophosphates,and phosphonamides.
A liquid detergent composition according to Claim 7 wherein the N-terminal endof said protease inhibitor is protected by a methyl, ethyl or benzyl carbamate[CH₃O-(O)C-; CH₃CH₂O-(O)C-; or C₆H₅CH₂O-(O)C-], methyl, ethyl orbenzyl urea [CH₃NH-(O)C-; CH₃CH₂NH-(O)C-: or C₆H₅CH₂NH-(O)C-],methyl, ethyl or benzyl sulfonamide [CH₃SO₂-; CH₃CH₂SO₂-; orC₆H₅CH₂SO₂-], and methyl, ethyl or benzyl amidophosphate [CH₃O(OH)(O)P-;CH₃CH₂O(OH)(O)P-; or C₆H₅CH₂O(OH)(O)P-] groups.
A liquid detergent composition according to Claim 8 wherein said peptidealdehyde is selected from the group consisting of: CH₃SO₂Phe-Gly-Ala-Leu-H,CH₃SO₂Val-Ala-Leu-H, C₆H₅CH₂O(OH)(O)P-Val-Ala-Leu-H, CH₃CH₂SO₂-Phe-Gly-Ala-Leu-H,C₆H₅CH₂SO₂-Val-Ala-Leu-H, C₆H₅CH₂O(OH)(O)P-Leu-Ala-Leu-H,C₆H₅CH₂O(OH)(O)P-Phe-Ala-Leu-H, and CH₃O(OH)(O)P-Leu-Gly-Ala-Leu-H.
A liquid detergent composition according to Claim 9 wherein said source ofcalcium ions is partially substituted with a source of another divalent ion.