text:
A chiefly send-off
Citizen Staff Reports 09/27/12
Henrico Fire Chief Edwin Smith retired Sept. 14 and received a surprise send-off from Henrico firefighters and county officials on his last day. Smith was greeted in the parking lot of the county's Public Safety building by several fire trucks, a ladder truck with an American flag flying and numerous members of the division, which he led for the past five years.
Smith spent 38 years with the division, beginning in May 1974 as a firefighter. He served in every suppression position as a firefighter and officer and advanced to serve as an inspector, investigator and later commander of the Fire Prevention section. A promotion to battalion chief and later chief of training for the division preceded his appointment to chief Aug. 18, 2007.
Smith is an active member of the International Association of Fire Chiefs, International Association of Black Professional Fire Fighters, Virginia Fire Chiefs Association and Central Virginia Fire and Arson Association. In 2010, he was appointed by Virginia Gov. Bob McDonnell to the Virginia Fire Services Board.
Business in brief
Citizen Staff Reports 10/23/2017 BusinessBusiness in BriefGeneral News
Henrico-based nonprofit Commonwealth Autism recently received the Standards for Excellence Institute's Seal of Excellence for successfully completing its accreditation program. Commonwealth Autism voluntarily opened itself to analysis by a peer review team during the last 18 months that examined the organization's compliance with the "Standards for Excellence: An Ethics and Accountability Code for the Nonprofit Sector." These standards cover areas such as: mission, strategy and evaluation; leadership – board, staff and volunteers; legal compliance and ethics; finance and operations; resource development; and public awareness, engagement and advocacy. Commonwealth Autism was one of six organizations in the Richmond region to be recognized and the first in the region to achieve full accreditation. In addition to this accreditation, Commonwealth Autism is recognized as an Accredited Charity with the Richmond Better Business Bureau and holds accreditation from the Code of Ethics for Behavioral Organizations (COEBO). > Read more.
Purify Infrared Sauna opens at GreenGate
Citizen Staff Reports 10/20/2017 BusinessGeneral News
Purify Infrared Sauna recently opened its second Henrico location at GreenGate Shopping Center in Short Pump.
Owner Mary Woodbridge opened her first Purify location on Patterson Avenue in July 2015. The new store is located at 301 Maltby Boulevard, Suite C, west of Short Pump Town Center. > Read more.
Henrico Master Gardener training program accepting applications through Oct. 27
Citizen Staff Reports 10/20/2017 General NewsGovernment
The Henrico County Office of Virginia Cooperative Extension is accepting applications for its next volunteer Master Gardener training program, which provides instruction in all aspects of horticulture.
Applications for the 2018 training program will be accepted through Friday, Oct. 27. Classes will be held from 9 a.m. to 12:30 p.m. Tuesdays and Thursdays from Jan. 16 through March 22. > Read more.
Henrico Schools to host Oct. 30 job fair
Citizen Staff Reports 10/20/2017 EducationGeneral News
Henrico Schools will host a job fair Oct. 30.
The event, to be held from 10 a.m. to 2 p.m. at the Fairfield branch library, is designed to attract potential full-time and substitute registered nurses, instructional assistants, bus drivers and school nutrition workers. > Read more.
Henrico Police to participate in 'Tip a Cop' Oct. 21
Citizen Staff Reports 10/20/2017 General NewsGovernment
Henrico County Police Division and the Virginia Division of Capitol Police are participating in "Tip-A-Cop" to Support the Special Olympics Saturday, Oct. 21.
From 11 a.m. to 2 p.m. and 5 p.m. to 9 p.m. that day at Red Robin, 11784 West Broad Street, members of the two agencies will be working for tips as a donation to the Special Olympics. > Read more.
Henrico Police to participate in 'Tip a Cop' Oct. 21
Participants sought for 'Walk to End Alzheimer's'
Fairfield meeting Oct. 25 to focus on cybersecurity
Music makers
McShin Academy expanding to St. Joseph's Villa
- More News
Print Edition
Oct. 19, 2017Current edition | Archive
Weekend Top 10
Chopt Creative Salad Co. opens Oct. 17 at Willow Lawn
Weekend Top 10
CSz Richmond to operate as nonprofit
Weekend Top 10
- More Entertainment
Obituary – Margaret B. "Peggy" Catlett
Obituary – Patricia Wells Melton
Obituary - Dorothy M. Wade
- More Obituaries
Green Top Outdoor Expo starts Saturday
Pearce Memorial Scholarship Craft and Home Show scheduled for Oct. 7
Lewis Ginter to host Fall PlantFest Sept. 15-16
- More Community
Work ethics: born in the crib (well, almost)
Growing a grown-up: Part II
How to grow an adult
- More Opinions
SunTrust to sponsor raceways's military appreciation program
Code Lacrosse joins Richmond Strikers
5 Henrico student-athletes win scholarships at awards dinner
- More Sports
|October 2017
Place an Ad | More Classifieds
CalendarLibbie Mill Library will host a Halloween Fair from 6:30 p.m. to 7:30 p.m. Enjoy storytelling, games, crafts and more. This free event is for families looking for a safe and fun indoor Halloween celebration. Costumes encouraged. Hosted by the Northside TAB. For details, call 501-1940 ext. 4 or visit http://www.henricolibrary.org. Full text


text: diagrammatical definition October 15, 2017 AZ Dictionary Link to this page adjective: diagrammatic shown or represented by diagrams diagrammatic shown or represented by diagrams View More Related Sources Definition for "diagrammatical" diagrammatic View More Sentence for "diagrammatical" Dad also got a large book… View More Equivalent for "diagrammatical" delineate View More How would you define diagrammatical? All the definitions on AZdictionary were written by people just like you. Now's your chance to add your own! Your Full Name Your definition Post Definition

subset: commoncrawl


text:
San Antonio Truck Accidents Caused by Driving Too Fast for Conditions
Call our San Antonio Truck Accident Lawyers at Carabin Shaw Law Firm if you, or someone you know has been hurt in a truck accident where the driver was driving too fast for the conditions on the road. It has been found that 23% of truck crashes happen when the driver is traveling too fast for the conditions. Such actions can have disastrous results.
When a driver is driving too fast for conditions, he/she is traveling at a speed greater than reasonable standard for safe driving. Conditions where drivers may drive too fast include: wet roadways (rain, snow, or ice), reduced visibility (fog), uneven roads, construction zones, curves, intersections, gravel roads, and heavy traffic.Failure to Avoid Driving Too Fast for Conditions
Truck drivers are supposed to employ techniques that allow them to avoid these conditions, but continuously fail. Failure to employ these techniques allows for accidents that cause serious injury or even death. Our San Antonio Truck Accident Lawyers are ready to fight for your compensation if you have been hurt as a result of this failure.
A truck driver is supposed to reduce their driving speed in adverse road and/or weather conditions and adjust to safely match the conditions on the road. Failure to do so causes more severe crashes. Recently, it was reported that 25 of speeding related truck accident fatalities happened during adverse weather conditions.
Truck drivers are also supposed to drive slowly with a loaded trailer. That is because large trucks with fully loaded trailers are 10 times more likely to roll over than those with empty trailers. A sudden speed adjustment may cause skidding or a rollover because of a shift in the load.
If you or a loved one has been injured in an accident with a large commercial vehicle or an 18 wheeler, call our San Antonio Trucking Accident Lawyers at Carabin Shaw Law Firm. Our dedicated team of lawyers know the federal rules and regulations that truck drivers, and the trucking companies that employ them, must follow. Call us at 210.222.2288, day or night, for more information.


text:
The ENO SingleNest Hammock Compare Price game of golf is the best source to receive oneself confronted with ENO SingleNest outdoor and utilize oxygen. The serenity during golfing is extraordinaire. Fresh air can provide relax plus the best strategy to boot away stress. Ability to hear the birds chirping each day will provide a great inner peacefulness which is impossible to describe. These can be easily made available from macizo trading post 40% away discount coupons ENO SingleNest which is not at all a herculean process, best gadgets can be quickly fetched against sierra trading post discount codes to enjoy the overall game to the maximum.
Composed of high-quality, breathable, and quick drying nylon, the ENO SingleNest Hammock is made for outdoor comfort. This hammock is light enough to backpack, but robust enough for backyard lounging. The attached compression stuff sack is a convenient way to store the SingleNest fast and easy. Heavy-duty triple stitched seams are strong enough to hold up to 400 lbs. Includes aluminum wiregate carabiners for easy attachment to solid anchor points and ropes, webbing, or whatever your strapping/connecting device of choice is during set-up (not included). Otherwise, using an ENO Suspension System delivers an extremely simple and streamlined set-up process (systems sold separately). Total weight: 16 oz. Folded dimensions: 3.5"Dia. x 4.5"L. Unfolded dimensions: 9'4"L x 4'7"W.
Don't like sleeping on the ground? Problem solved! The ENO Single Nest Hammock is lightweight, compact, durable, and a piece of cake to set up. Made with breathable nylon micro fiber fabric that is UV treated and mildew resistant, this hammock can hold up to 400 pounds and packs into its built in stuff sack the size of a grapefruit. This great piece of gear uses nautical grade cord on the ends to hang and includes 2 stainless steel carabiners. For a quick and tree-friendly hanging option, use ENO Slap Straps or ENO Atlas Suspension System. So durable it comes with a 2 year warranty.


text:
American International GMK432EX1 Dash Kit
2000 Pontiac Grand Am
Crutchfield Customer Review (What's this?)
Fit right into the dash of my 2000 Grand Am GT. Easy, peasy my 9 year old could have done it.
Pros: Come's to you and a single package, like a model car/plane from the day's of past. Twist off the pieces you need and go for it.
Cons: Identifier's need to be marked better on the piece's involved with the install.


text:
read time:
After your ride & drive program wraps up, you can just feel the weight lift off your shoulders. However, before you get too relaxed there is still critical follow up work to be done after the ride & drive concludes. We have identified 3 post - ride & drive best practices you should follow to help maximize engagement and CRM accuracy.


text:
Thanks Samantha!
From: gsis_admins-bounce@xxxxxxxxxxxxx <gsis_admins-bounce@xxxxxxxxxxxxx> on
behalf of Samantha Mimbs <samantha.mimbs@xxxxxxxxxxxxx>
Sent: Thursday, August 27, 2015 7:24:20 PM
To: gsis_admins@xxxxxxxxxxxxx
Subject: [gsis_admins] GSIS Board Meeting Location
Meeting to be held on September 29, 2015 at 10:00 am. It will be held at:
25 Brooklyn Avenue
Forsyth, GA 31029
Best Regards,
Samantha Mimbs Data Coordinator, Monroe County Schools
Phone: 478-994-2031<tel:478-994-2031>
Email: samantha.mimbs@xxxxxxxxxxxxx<mailto:samantha.mimbs@xxxxxxxxxxxxx>
Website: http://www.monroe.k12.ga.us<http://www.monroe.k12.ga.us/>
Address: 25 Brooklyn Ave Forsyth, GA 31029
IMPORTANT: The contents of this email and any attachments are confidential.
They are intended for the named recipient(s) only. If you have received this
email by mistake, please notify the sender immediately and do not disclose the
contents to anyone or make copies thereof.


text:
Let's get to the breakdown...
Promotional Images
Sultry Cranberry Picture Tutorial
That Girl Fever Shadow and Luminous Contouring Palette
The first thing you notice about these palettes are obviously the packaging. I love how slim these are, which makes them nice and compact. Both palettes are accompanied with a massive mirror, which is always a plus. Also, I appreciate that the palettes are different colors. This is helpful when you're rummaging through your makeup collection! You'll be able to tell the difference between the palettes without having to open them.
The eyeshadow palette contains 9 different eyeshadows. They're nice sizes, at about 3/4 of an inch, and beautiful. They swatch wonderfully and look super pigmented on the eyes. I've done a few looks with them (see the one above!) and they're great! The only ones that fall a little short for me are "#Just 10 Seconds" and "#All That Glam". They're a bit chunky and have a bit of fallout to them. The other eyeshadows are nice, smooth, and opaque :) I'm actually super impressed with this palette and right now it's $33!
Next, let's take a look at the contouring palette. Contouring has been all the RAGE in Korea right now, even though it's been popular here in the states for a while. However, instead of chiseling yourself new cheekbones, it's more popular to contour along your jawline in Korea. However, we all know the importance of finding a contour that is not orange or too grey so that the contour looks natural. I actually think Pony did an excellent job with this! I also love that there's two shades of contour, so depending on your skin tone or how heavy of a contour you'd like, you can definitely customize your look. However, the highlights are a bit lack-luster for me. I can see how they're a "natural" highlight, but they're just not exemplary highlights. They're not super shimmery and there's nothing super "glow-y" about them. Still not terrible though!
Pony Effect That Girl Outfit Lipsticks
I have to say, I really like these lipsticks. The packaging is gorgeous and I love how bright and happy they are. The lipsticks have a magnet inclosure, so they snap shut when you close them. They seem pretty secure too! The formula of these are nice and smooth and don't take a ton of swipes to look nice and opaque. These are a satin finish and I don't find them super moisturizing. One this is.. I think these are a bit over-priced at $18 a pop (regularly $22) considering there's nothing phenomenal or super luxurious about the formula. I mean, I REALLY like MAC lipstick formulas and they're cheaper than these.
Pony That Girl Outfit Lipstick in "Noble Brunch"
This is easily my favorite product out of this collection. It's the PERFECT nude! If you have had issues with finding nudes that don't make you look dead, your search is over! It's the perfect peach-y, pink nude that will go great with any look. A great color to through in your purse and wear daily! If you're going to get one lipstick.. get this one!
Pony That Girl Outfit Lipstick in "Expensive Dinner"
I'd consider this a very wearable red. It's a bit of a cranberry red color and doesn't have any tones of orange to it. I like it and think it would be a nice color for the upcoming holiday season :)
Pony That Girl Outfit Lipstick in "Vibrant Club"
I noticed in the promotional pictures that this looks really purple. However, I'd say again, this is a wearable berry color. It has a nice hint of deep plum. In my opinion, berry red is a good description of what this color looks like in person!
That Girl Illusion Nail Laquer
Now these are the only products from the line that I haven't tried yet. They look gorgeous though and I'm super excited this set comes with a cellophane effect polish. It's been so trendy in Korea and I can't get enough of it. Plus, look at those other colors! How pretty and I have nothing remotely similar to these shades so YAY! :)Well guys, I hope you're excited! If you're a fan of Pony, like I am, I don't think you'll be disappointed with the collection. Plus, it's just in time for the holidays! :)


text:
Maxine Waters Reclaiming Her Time Just Became Everyone's New Summer '17 Mantra
California Congresswoman Maxine Waters puts her foot down by reclaiming her time.
(Photo: The Jacka via Instagram)
Bay Area rapper The Jacka was shot and killed in East Oakland Monday (Feb. 2) night, CBS San Francisco reports. He was 37.
Jacka, whose birth name is Dominic Newton, was shot in the head, according to SFGate.com. Witnesses say he was gunned down on the 9400 block of MacArthur Blvd. around 8:15 p.m. He was pronounced dead at Oakland's Highland Hospital.
Authorities have yet to officially announce him as the shooting victim, although a reporter from the Bay Area's KPIX News confirmed the fatality. The gunman and motive remain unknown.
By late Monday night, dedications were already pouring in for the fallen rapper via social media. Bay Area native G-Eazy tweeted, "Damn... RIP @theJacka the Bay Area lost a legend," while Fashawn mentioned collaborating with Jacka. "It was an honor to work with you and build before you left king," he tweeted. "The coast lost a real one." Nipsey Hussle posted a few tributes to Jacka, writing next to one video,"His city Loved him. My n***a rep the bay well #RESTEASY"
Born in Pittsburgh, Calif., Jacka began his rap career in the late '90s as a member of the Mob Figaz crew. He left the group to go solo in 2001, putting out a number of studio albums, mixtapes and compilations over the last several years, including 2014's What Happened to the World, and a joint LP with Freeway titled, Highway Robbery.
His shooting marks the 11th homicide in Oakland since that start of the year.
BET.com is your #1 source for Black celebrity news, photos, exclusive videos and all the latest in the world of hip hop and R&B music.
Click here to subscribe to our newsletter.
California Congresswoman Maxine Waters puts her foot down by reclaiming her time.
"Black women: Be fearless. Speak out for equal pay."


text:
Telling Thoughts
A gentle reminder to those who suffer from conveniently short memories. Let them remember that it was the Bush / Cheney Republican administration who started the two current wars, first in Iraq and then in Afghanistan, years later, still being fought with no end in sight, at a cost of more than 4000 American lives, and hundreds of billions of dollars and counting, after first having lied to the world about the existence of weapons of mass destruction in Iraq, as an excuse to invade.
The Bush / Cheney Administration fully approved of the way that Wall Street operated in a deregulated market, bought about by the administrations of Reagan and Clinton (Republicans still support the failed concept) without conceding, that it was millions of corruptly packaged and marketed sub-prime housing loans that were primarily responsible for the collapse of the global financial system. The rest of the world also suffered as a result.
Surely it is time for America to start looking after the interests of its own people. It must be careful to avoid taking the wrong path being traveled by Republicans. A path proven to be strewn with tragedy, human loss and financial hardship. Barack Obama inherited a country devastated by Republican madness and maladministration. No foreign enemy could have inflicted the level of damage to the country and it's reputation as that delivered by Bush / Cheney and the Republicans. Now as President Obama struggles with the horrendously difficult task of righting the America's ship of state, he will need strong support both in the house and senate to get the country back on it's feet.
Evan Bayh's retirement is a good thing.
As a less than loyal Democrat, Evan Bayh could not be relied upon to provide the support required to pass the President's campaign agenda when it counted. In many of his positions, he looked more like a Republican than a Democrat. War mongering Republicans have had their chance, and in the process they destroyed the economy and tarnished the country's reputation with their flawed ideas. If Americans want a decent future they need to support Barack Obama in his efforts to get things back on a sound footing once again.
It will take many years, but it can be done,
Comments are closed.


text: I have an idea. How about we get up with a bunch of bloggers, and do a Christmas program? As often as you can, every day if you can, do a Christmas/whatever holiday you celebrate post. Whether it's a DIY or a photo story, anything counts! And if you are not a blogger, you can email it in. I cannot do it because I have a special email account where the adult has to register. But maybe, my dad will do it? Anyway, who's with me? I will make a special pin thing. If you are, post about on your blog, and tell your friends! It starts on December 1st.


text:
As you well know, quilting on a sewing machine can be hard on the eyes. If the lighting is not just right, it can be difficult to work for long periods of time without your eyes becoming tired and strained. Not only is sewing in poor light difficult, but so …Read More »
Home / Product & Supplies / Lighting and Magnifers


text:
- Drug Transport, Inc DTI Decieved Carriers Tucker Georgia
- DTI logistics Drug transport bad lease program for truck drivers ran a whole month for free 2800 miles company will lie to you they have to many deductions run from this they want you to pay for everything they have no expence tucker, Georgia
- Muna Trying to collect on a case that was closed by the company I had a contract with. Tucker Georgia
- Westside Transport, Cedar Rapids, IA This is a terrible company. I would not refer anyone to go to Westside. Cedar Rapids Iowa
- Netspan Group Educational Testing Services led me to believe the Post Office was hiring and I needed to go ahead and study because I would be testing very soon,I was unemployed and desperate for a job,now I cannot get my money back. Tucker, Georgia
- Richard Tucker - Rick Tucker Lied about working out payment plan. Cashed checks without providing services. Was suppose to file divorce for me. Bonsall California
- Bill Tucker - William Tucker - Tucker Associates - Tucker Insurance Systems opportunity scam Celebration Florida
- MSW Industries.com LLC Arthur Tucker Arthur Tucker/Arthur M. Tucker/MSW Industries.com LLC/Mel Knox/MSW INVESTORS CORP (AKA-MIC) Scammed, strung along filled with false hopes, false promises, elusiveness and deceptiveness Los Angeles California
- Art Kassel, Detective Training Institute, and DTI students should take the time to see they are illegally using the BBB reports Ripoff San Clemente California
- Bentley Inc. Bentley corp. , Bentley Inc. Transport, Bentley Transport Inc. Haven't been paid for work done. They say I lost the paperwork but it was sent or dropped off when and where I was told to. Cheyenne Wyoming
- Tucker Capital - Ripping Off to Truckers with Junk Trucks Forest Virginia
- Report: #1052359
Drug Transport Inc.
Drug Transport Inc.1939 Forge Street, Tucker, Georgia USA
Drug Transport Inc. DTI was I was wrong for letting me down.I was forced to leave company. Tucker Georgia
REBUTTAL BOX™ | Respond to this Report! | Consumer Comment
Are you an owner, employee or ex-employee with either negative or positive information about the company or individual, or can you provide "insider information" on this company?
This is the best way to manage and repair your business reputation. Hiding negative complaints is only a Band-Aid. Consumers want to see how a business took care of business.
All business will get complaints. How those businesses take care of those complaints is what separates good businesses from the rest.
Consumers love to do business with someone that can admit mistakes and state how they made improvements.
Corporate Advocacy Business Remediation and Customer Satisfaction Program.
A program that benefits the consumer, assures them of complete satisfaction and confidence when doing business with a member business.
Show customers why they should trust your business over your competitors...
Ripoff Report
About you?
Ripoff Report
A business' first
line of defense
on the Internet.
If your business is
willing to make a
commitment to
customer satisfaction
Click here now..
Does your business have a bad reputation?
Fix it the right way.
Corporate Advocacy Program™
Set the record straight:
Arbitration Program
Hello. My name is hidden for privacy use. I would not refer the most worst driver to work for Drug Transport.
The truckload division is on Stone Drive in Tucker,GA. The local truck division is at Forge Road in Tucker,GA
Crazy a*s dispatchers and bad trailers. Trailers stay in bad shape.. You will get paid every two weeks if you work the local job and you will make 14 bucks per hour. You can make better money working in a warehouse getting paid every week.
Wrong for this trucking company to pay local drivers every two weeks. The company have a high trunover rate and the drivers who work for DTI/ Drug Transport always saying bad stuff about the company. The drivers were trying to run me away when I got hired. The stuff drivers said was true. I had to see for myself. The company is bad with a high turnover rate.
The dispatchers are rude and crazy. They will call your phone too many times in one day while you are out focusing on driving and taking care of the customers. DTI have one male dispatcher. A black american dispatcher who is rude and he take his personal problems out on drivers. The guy needs to see a counselor. He needs real help. He will make you look bad and he will mess up your driving record and he likes to tell lies to his boss to make himself look good. He don't know how to
plan trips and he don't care if drivers are driving illegal. He like when drivers are breaking DOT rules. I am for real.
You will have to unload all boxes inside the trailers all the time. 90 percent touch freight. You will only get 60 bucks to unload a full 53 feet trailer at all Marshall and TJ Maxx stores. All the trucks are leased thru Rider Company.
If you mess up anything on a trailer you will have to pay for it out of your check. They will even charge you for a flat tire.
I was charged 350 bucks for the hinge on a trailer door. Sad as hell.
No prepass or easy pass. No Qualcomm. All work conversations and load assignments are done by phone only.
You will get home weekly. Thats the only good thing about the OTR division.
One dispatcher quit in January. I heard that he quit because the company was not treating him right.
The boss made me quit because he did not want to have termination written on my seperation notice. I was forced to resign because of the crazy dispatcher that works for the company. He needs special help.
Thanks for reading this report. This is a true report and any driver can witness that reports made against Drug Transport is true. I would refer drivers to work for the company. Heck no I wouldn't do it.
This report was posted on Ripoff Report on 05/20/2013 11:45 AM and is a permanent record located here: http://www.ripoffreport.com/reports/drug-transport-inc/tucker-georgia-30084/drug-transport-inc-dti-was-i-was-wrong-for-letting-me-downi-was-forced-to-leave-compan-1052359. The posting time indicated is Arizona local time. Arizona does not observe daylight savings so the post time may be Mountain or Pacific depending on the time of year.
Ripoff Report has an exclusive license to this report. It may not be copied without the written permission of Ripoff Report. READ: Foreign websites steal our content
Click Here to read other Ripoff Reports on Drug Transport Inc.
If you would like to see more Rip-off Reports on this company/individual, search here:Search Tips
In order to assure the best results in your search:
- Keep the name short & simple, and try different variations of the name.
- Do not include ".com", "S", "Inc.", "Corp", or "LLC" at the end of the Company name.
- Use only the first/main part of a name to get best results.
- Only search one name at a time if Company has many AKA's.
ZipBooks Accounting Software
and ZipBooks for Invoice Templates
Advertisers above have met our
strict standards for business conduct.


text:
The Department of Geology at Stephen F. Austin State University (SFA) invites applications for a tenure-track position at the assistant (or associate) professor level. Applicants must have a doctoral degree in geology or a related field with emphasis on petroleum geology, a strong commitment to excellence in teaching and a willingness to direct Master of Science geology graduate students in research. Preference will be given to candidates with petroleum industry and/or research experience. Teaching responsibilities will include introductory courses, upper-level and graduate courses in the applicant's specialty, and occasional weekend field trip courses. Other expectations include research, university service and continuing professional development.
Submit a letter of application, CV, Transcripts, Statement of Research Philosophy, Statement of Teaching Philosophy and contact information for three references to https://careers.sfasu.edu (posting 0605053).
Also mail official transcripts to:
Dr. Wesley Brown, Search Committee Chair
Stephen F. Austin State University
Department of Geology
PO Box 13011 SFA Station
Nacogdoches, TX 75962-3011
(936) 468-3701
Review of applications will begin on November 3, 2017, and continue until the position is filled. SFA is an equal-opportunity employer. This is a security-sensitive position and will be subject to a criminal history check.


text:
|» Products
|» Coffee Table
|» Coffee Table
Coffee Table
Coffee Table
Product Code : 15
Product Description
Based on our industry experience we are manufacturing and supplying quality tested range of Coffee Table. Offered range is specially designed to enhance looks of cafeteria. Our tables can be used in cafeteria and canteens of schools, office and restaurants. Entire range is manufactured by using high grade raw materials sourced from leading vendors of market. Clients can buy this Coffee Table from us at best of the market price with commitment of timely delivery.
- Light weight
- Easy to clean
- Long life
G-1001, Phase III, Sitapura Industrial Area, Jaipur - 302022, Rajasthan, India
Mr. Dhiraj Patel (Proprietor)


text:
Consumer Electronics Reviews
MSRP: $100.00at
Bottom Line: Everything's better about this year's Amazon Echo, except for the sound quality. But it's a lot easier to swallow given the much lower price.Read Full Review
MSRP: $200.00at
Bottom Line: Harman Kardon's Invoke, the first Microsoft Cortana-powered speaker, is a poor competitor for voice assistants from Amazon and Google.Read Full Review
MSRP: $299.00at
Bottom Line: The Lumu Power is an accurate light meter with a clean, modern app interface, but it needs an iOS device with a Lightning port to work.Read Full Review
MSRP: $599.00at
Bottom Line: The Fujifilm Fujinon XF 18mm F2 R is showing its age, but is still a good choice for X shooters who prefer lightweight lenses.Read Full Review
MSRP: $999.00at
Bottom Line: If you don't mind spending the money, the Fujifilm Fujinon XF 16mm F1.4 R WR lens will wow you with its f/1.4 design and wide-angle field of view.Read Full Review
MSRP: $399.00at
Bottom Line: The Fujifilm Fujinon XC 50-230mm F4.5-6.7 OIS II is an affordable telezoom for the X system, but its narrow f-stop limits use in dim light.Read Full Review
Bottom Line: The Eufy Lumos is a standalone Wi-Fi smart bulb you can control from anywhere using your phone. It works without a hub, and at $20, it's a good way to try out smart lighting.Read Full Review
MSRP: $100.00at
Bottom Line: Roku's top-end 4K media streaming box, the Ultra, is packed full of handy features, but whether they're worth the $100 price depends on your viewing needs. For many, the $70 Streaming Stick+ will be just fine.Read Full Review
Bottom Line: Cuphead pays homage to old school animation, as well as side-scrolling video game shooters of old. While it lacks the stage-heavy mayhem of other games in the genre, Cuphead more than makes up for it with its vicious boss battles.Read Full Review
MSRP: $199.00at
Bottom Line: The Sonos One is a versatile wireless speaker with excellent sound quality, support for multi-room audio, and Amazon Alexa voice control.Read Full Review
MSRP: $95.00at
Bottom Line: With its new cotton hoodie, Scottevest takes its original tech-friendly, pockets-a-plenty shirt and makes it warmer to wear while adding even more storage.Read Full Review
MSRP: $599.00at
Bottom Line: The mirrorless Fujifilm X-A3 camera and the images it captures look great, but its autofocus system is way behind the competition.Read Full Review
MSRP: $1,299.00at
Bottom Line: The Canon PowerShot G1 X Mark III is smaller than the Mark II, despite having a bigger sensor, but sacrifices zoom range to get there.Read Full Review
MSRP: $1,000.00at
Bottom Line: Panasonic's top-end 12-60mm zoom lens delivers image quality that lives up to its Leica branding.Read Full Review
MSRP: $139.00at
Bottom Line: Bell Robot's Mabot modular construction system is an accessible way to teach robotics and programming to kids.Read Full Review
Bottom Line: Fire Pro Wrestling World is a Steam Early Access project, but its combination of classic gameplay and contemporary PC game features make it a promising wrestling title even in unfinished form.Read Full Review
MSRP: $2,000.00at
Bottom Line: The Panasonic Lumix DC-GH5 is the mirrorless camera to get for 4K video, but if you're more concerned about stills, it's not a clear-cut winner.Read Full Review
MSRP: $4,000.00at
Bottom Line: LG's 65SJ9500 is a pricey 4K LCD TV that boasts picture quality and style that almost puts it in arm's reach of OLED models.Read Full Review
MSRP: $200.00at
Bottom Line: The Shield TV is a powerful Android TV-based media streaming microconsole with plenty of gaming tricks and hands-free Google Assistant support to help justify its hefty price tag.Read Full Review
MSRP: $49.00at
Bottom Line: The Google Home Mini smart speaker takes on Amazon's Echo Dot with a cute, fabric-covered design and Google Assistant smarts, but its lack of an audio out is a major bummer.Read Full Review


text:
Tart and Tangy BBQ Cranberry Sauce
We're excited to share a new family favorite recipe with you for Cranberry BBQ sauce. We think your family will love it too because this recipe delivers a terrifically tangy, sweet heat. Its layers of flavors will make your ribs and chicken extra finger-licking good. Plus, homemade BBQ sauce is a simple way to make your guests feel special. We love using bold flavored cranberries that stand up to barbecued meat. Plus, cranberries are America's Original Superfruit and have many wonderful health properties. They contain antioxidants; are naturally fat and cholesterol free, low in sodium and are a good source of dietary fiber. Be sure to check out UScranberries.com to view more amazing recipes. Please enjoy our recipe below!
Cranberry BBQ Sauce
- ½ cup fresh or frozen cranberries
- 6 oz. tomato paste
- ½ cup orange juice
- ½ cup apple cider vinegar
- ¼ cup brown sugar
- 1 tsp. paprika
- ½ tsp. cayenne pepper
- Add all ingredients into a non-stick saucepan.
- Heat on medium until mixture starts to simmer and then mix to combine ingredients.
- Cover and continue heating until mixture comes to a boil, turn down to a simmer and stir mixture. Continue simmering, stirring intermittently until sauce thickens, approximately 10 minutes.
- Remove from heat and allow to cool. When mixture is cool, blend with an immersion blender until smooth.
- Pour into covered container and refrigerate until ready to use.
Party Bluprints is proud to have partnered with the US Cranberry Marketing Committee to develop recipes and promote the goodness of cranberries.


text:
Welcome to ABC Radio National. Skip to:
RN Navigation
RN Search
RN Content
Search RN
View by
When Peter Bush chooses Spanish works to translate he starts at the top. He's just finished translating the 15th century Spanish masterpiece by Fernando de Rojas called Celestina.
Now for some craic with words, as they say in Ireland. Craic is how the Irish describe good times, but 'craic' isn't actually an Irish word. It was borrowed from the English, but having craic or simply 'the craic' has become more a part of Irish language and culture. So will the word craic be included in the Historical Dictionary of the Irish Language?
This [series episode segment] has
Is translation the art of failure as Umberto Eco said? Five Australian authors have had their works translated into French by Mireille Vignol: Peter Temple, Kate Grenville, Anna Funder, Sylvia Lawson -- and six books by Kenneth Cook.
Although biblical in origin, the term 'ivory tower' is now synonymous with academic elitism.
This [series episode segment] has
Australia is in the midst of a languages crisis: too few Australians are enrolling to study languages other than English.
One of the longest epic fantasies in the world is the 19th century Urdu classic Hoshruba. The huge work was compiled by two of Urdu's greatest prose writers. Spread over 8,000 pages, it was immensely popular and acclaimed in the history of Urdu literature.
Twelve years ago, the state library of New South Wales received a treasure trove of Spanish literature. More than 1,100 editions of the Cervantes novel Don Quixote and hundreds of books devoted to the influence of the 17th century Spanish writer were donated to the library by a local Sydney doctor. We take a tour of the Cervantes Collection.
You may have seen the musical of Les Miserables or perhaps watched the film with Gerard Depardieu, but have you read the book? Victor Hugo's Les Miserables has been given fresh life in a new translation by Australian Julie Rose.
This [series episode segment] has
image, and
'A howl, a shout, or a machine-gun, or the wind or a wave', this is how Caribbean poet Kamau Brathwaite describes writing in the vernacular.
This [series episode segment] has
It's believed to be the world's first modern novel, penned a thousand years ago. The Tale of Gengi by Murasaki Shikibu starts when a woman of lower rank in the court gives birth to a son called Genji. He is favoured by the emperor because he's so beautiful, talented and likeable. He goes on to have many love affairs, which allows Murasaki Shikibu to explore ideas of love, court politics, friendship, life and death. Ten centuries on, the popularity and influence of this ancient text has not diminished and this year to honour the millenium of The Tale of Gengi, Japan is celebrating.
The Book Show is an archived program which is no longer broadcast
Find out more
The Book Show's high profile guests reveal their top reads.
Find out more
More Book Show features


text:
I had a brief encounter recently where someone shared how deep their hurt is and that no one else has had such a painful existence as they. I took the opportunity to express that we all go through hard times. Life has its ups and downs and no one is exempt. Some of the painful times are harder than others but learning how to handle it and how to rise above it is what sets some people apart. Those who look like they have it all together have only learned how to get up and get going, moving past the circumstance, and do what needs done.
Sometimes pain can envelop your mind and you can't see past the storm, but you are not alone. The Bible is full of people who experienced pain like yours and worse.
That is one reason we have the Bible. So we can read it and learn from it.
The painful times, the trials, the rejection, the rumors, the hard times of life...God uses these times to make us stronger if we will let Him. Sometimes the biggest lessons come from the most painful times.
There were times in my own life when I felt the world was against me. We ministered in several difficult churches and there were days I wanted to walk away from ministry and have nothing to do with church people or the church. I've even expressed to my husband that when he retires I don't think I want to go to church anymore. Because people are cruel.
However, if I had not gone through those times I wouldn't have the insight I now own. I wouldn't have the backbone to say what needs to be said. I wouldn't have the wisdom that reminds me to not say some things. I wouldn't have the freedom to be myself because I would still be caught up in trying to please everyone.
You are not alone. We have all been there. You will get through this.
Get out your Bible and start reading.
Read about Job. If anyone had reason to be in misery he did, but he never lost faith in God.
Read the Psalms, they are eye opening.
Every hero of the Bible went through difficult times.
And you will too.
It's part of life but you will make it through if you learn to trust in the Lord in every area of your life.
Pray for wisdom. Pray for peace in your thoughts. Pray for guidance.
Pray for discernment. Pray for joy.
He will provide it all. Maybe not today, but be patient in the trial.


text:
As you may know, there was a bomb that went off in the foyer of the MEN arena in Manchester after the Ariana Grande concert on Monday.
Read More About What Happened
So many innocent people died and even more were injured. These people were concert goers or those who were picking up someone. This should of been an evening filled with joy but it has ended in disaster.
This is nobody's fault apart from the people involved with the bombing. It's so sad but it's amazing to see how everyone has been united despite this tragedy.
I can only give my consolodences to those affected by this incident. My heart truly goes out to them all.


text:
All our items are custom orders and therfor bespoke these will be delivered by Hermes or Royal Mail free within the UK
Outside the UK all items must be signed for on deilvery and you will be suppled with a tracking number
All orders are delivered boxed and garments are wrapped in acid free heirloom tissue , this is to ensure they can be kept for years to come and can be handed down
Included are care instructions and if possible spare button and yarn
If you see a garment on our site please ask us to custom knit for you
To make a custom order please contact me via the contact page
These orders must be paid for in advance as a purchase
I will keep clients updated at all times regarding their order
and when it is ready to ship
As your order is a bespoke item you waive the right to cancelation rights
Custom orders are knitted on a first come basis and i will endevour to get all items out on a timely basis
Postal charges
UK postal is free please note this will not be inasured or signed for and are posted at buyers own risk
Our preferred method of postage is set out below
UK Signed for delivery £8.00 Additional Items £1.50
Australia £18.00 additional items £2.50 per item
EU £11.00 additional items £2.00 pe item
France £9.00 additional items £1.50 per item
USA £19.00 additional items £2.50 per item
Customs and import taxes
Buyers are responsible for any customs and import taxes that may apply. I'm not responsible for delays due to customs.
Custom orders may not be returned
All other garments, we only accept returns if goods are faulty
We ask that you measure or to give us the garment size required, we are not thefore responsible if the garment is the wrong size
Goods that have been worn or washed will not be accepted as returnable
Any return postage shall be paid by the customer


text:
Comment Re:Health Data is owned by the Patient (Score 3, Informative) 99
A few years ago I was in the position of changing doctors and had a very hard time getting my data from my old doctor to the new. I had to constantly go after them to get copies. They kept putting it off and at one point said they owned the records since they prescribed any and everything medical for me. I explained that I owned my records, they may own the paper printed but the information is mine. I bought and paid for the tests and I paid the doctor to decide what tests needed to diagnose my problems and issues. I'm even paying them to keep my records safe by going to them all the time , being my primary care physician. As long as I am a customer of theirs they are happy with keeping my records on file and "lending" out as needed to doctors they refer me to but as soon as I said I was changing doctors (I moved and wanted a closer doctor then the 1.5 hour drive it would now take to get to my old one) they got all up in my face over my records.saying they owned them. After 3 months I finally got my records which really pushed me back 3 months on any medical diagnoses and treatments since the new doctor didn't want to start or stop anything the old doctor told me to do since he didn't have any facts of what my history was at the time. So now any tests I have done I make sure I get a copy for my own records and keep them in my safe at home.


text:
Personal Information:
|Postdoc researcher
|Phone or fax:
Three interconnected themes run through my research: brains and cognition, ecological niche and adaptive flexibility (an ability to develop novel solutions to problems, through processes including innovation and social learning). These themes are important throughout the history of the human lineage as well as for understanding the origins of Homo sapiens. In order to understand how these characteristics changed and interacted throughout human evolution, I focus on relevant case studies from a range of different Palaeolithic periods and regions. This has varied from a study of the role of adaptive flexibility in the earliest hominin expansion out of Africa (in my doctoral research), to Neanderthal learning of subsistence skills (in a postdoctoral project at the
A key part of my current research focuses on the earliest occupation of north-west


text: In your message about "Re: Inkjet printing.", you said: > >>I would agree - generally, 20# for laser printers, although I've done card >stock successfully too. But this textured stuff is inkjet-rated by Paper >Direct, and it's got to be at least 80# cover stock. The smoother the texture the better for laser printing; the only issue with weight is whether it will run through the printer's paper path. Since my printer has a straight-through option, I rarely worry about it. --<<<<<<<<<<<<<<<<<<<<<<<<<<<<<>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>> Robert Goff, Head Scribe and Vellum Scraper, The Outlaw Press (500)447-0070 - (505)255-9801 (fax) rgoff@xxxxxxxxxxxxxxx - http://www.outlawpress.com/outlaw/ <<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>


text:
Secrets in Temple RunFree
This app is to reveal all the secrets in Temple Run. So how can you miss this Temple Run Secrets app if you are a fans of Temple Run!!!! Come and download this Temple Secrets app, and you will learn many tips and secrets in Temple Run. There is also guides for you to become the pro of Temple Run!!!! This guide is an unofficial version and is not endorsed by or affiliated with the creator of this game or its licensors. This application complies with the US Copyright law guidelines for fair use. All characters, their names, places, and other aspects of the video game described within this application are trademarked by their respective owners. This application does not copy any portion of the game, nor does it contain screenshots of the game, only original text descriptions. If you feel there a direct copyright or trademark violation that does not follow within the fair use guidelines, please contact us directly to discuss.
Downloads: 2,550+ (for Android)Report app for spam or abuse!
Recommended apps
Super File Manager freemium Free
Free, Safe, Small, Simple. One of the best File Manager apps on GetJar.
GTA 5 APK for Android and iOS Free
Download GTA 5 apk file by clicking the download button given below If you are downloading the file
GTA San Andreas FOR android Free
Gta San Andreas System Requirements Following are the minimum system requirements of Gta San Andrea


text:
A master's degree is a postgraduate academic degree. One must already have an undergraduate degree to apply for a master's program. Most master's degree program would require students to complete a master's thesis or research paper.
Generally speaking, a human resources degree refers to the workers that make up an organization and, therefore, prepares students for different kinds of jobs. While working toward a degree of this kind, students may take classes that delve into topics like pay equality, employment law, organizational behavior, leadership and management.
Spain, officially the Kingdom of Spain, is a sovereign state and a member state of the European Union. It is located on the Iberian Peninsula in southwestern Europe. Spanish universities regulate access to their own degrees and they fix the academic fees. They can also offer unofficial postgraduate degrees. The capital city Madrid has possibly the largest number of bars per capita of any European city and a very active nightlife.
Salamanca city is the capital of the Salamanca Province, Spain. It is a metropolitan area with over 200,000 residents. It has several buildings that are part of the many sights and sounds of the city. Among these are church and university buildings.
Request Information Master's Degrees in Human Resources in Salamanca in Spain 2017/2018
or search for similar programs here


text:
My Content
Membership has its privileges! Sign up or sign in to gain full access to both All About Jazz and Jazz Near You. Learn more.
Giorgos Kontrafouris was born in Athens, Greece in 1967. He studied classical piano at the National Conservatory in Athens. At the age of 16, he turned his interest to jazz music and he followed up jazz improvised lessons with Markos Alexiou. He studied also with Jim Beard and Jarmo Savolainen. He holds a Bachelor's degree and a Master's degree in Jazz Performance from Sibelius Academy in Finland.Read more
Develop a column, write album reviews, cover live shows, or conduct interviews.


text:
"Debate is healthy and no one in this chamber-starting with me has a monopoly on being right."
You may be left or right, you may be Gandhian, Nehruvian, Marxist or follower of your own path of thinking.. But you must have a vision on how to take the country along the lines of development. Start brainstorming, polish your ideas and Wrangle-on.
Topic: Model Village vs Smart cities: the way forward for development; Difference of Approach or battle of ideology
Participants will be given 5 minutes to put forward their ideas and 3 minutes for open discussion. You will be judged on the basis of your rational/critical thinking, persuasion capabilities and oratory skills.
Note: Language cannot be a barrier for knowledge. Participants can choose either Hindi or English as their medium.
Coordinator – Rajat Kr. Kanaujia 9455455766
Co Coordinator – Akshay Nayak 9450979147
Rajiv Gandhi Indian Institute Of Management (IIM), Shillong
Indian Institute Of Management (IIM), Kashipur
Indian Institute Of Management (IIM), Kashipur
Indian Institute Of Management (IIM), Kozhikode


text:
Business letters
Ending the shambles at Heathrow
LIKE you, I cheered when the Competition Commission called for the break-up of BAA. However, your editorial last week talked about the "new owners of Gatwick and Stansted", which implies you think BAA will keep Heathrow. I think it essential that BAA is required to sell Heathrow. It has demonstrated over several decades that it cannot manage the airport.
Martin Budden Richmond
WHEN I was returning from Switzerland at the end of July, there were only two or three officers checking passports at Heathrow's Terminal Five. As a result, we had to wait in long queues at passport control.
BAA get its act together? Not only BAA but also the government. It would appear from our experience (not for the…


text:
Ex-cricketer Narasimha Rao in Hyderabad yesterday
Hyderabad: Former Test cricketer and legendary batsman VVS Laxman's childhood coach MV Narasimha Rao lost his prestigious medal, which was awarded to him by Britain's Queen Elizabeth in 2011 for his work in setting up cricketing structure in Ireland.
Rao, under whose leadership Hyderabad won their only Ranji Trophy title by beating Delhi during the 1986-87 season, had been to a function with the medal, but after returning home, he found it missing from the bag.
"I went to a college function as chief guest. The medal was showcased there, but I don't know how the medal (left) got misplaced," the 62-year-old Rao told mid-day yesterday.
Rao, who played two Tests as an all-rounder from 1978 to 1979, went to Britain after cricket retirement where he did a lot of charity and anti-racism work apart from his cricketing contribution to the nation.
"It's as good as a Padma Shri for me and receiving it from the Queen makes it more prestigious.
"They gave it to me for my contribution in setting up cricketing structure in Ireland. I appeal to everyone, whoever gets any news of this medal, please inform me," said Rao, who is now the Director at Hyderabad Cricket Association's Academy of Excellence.


text:
One hundred year old Nellie Slinger claims to have discovered the secret of old age - never get married.
Nellie, who celebrated her birthday yesterday, said dumping a cheating fiance made her determined to enjoy a long life.
"Not having anyone to run around after had something to do with it," she said.
Nellie had dated her fiance for two years before the Second World War when her factory workmates saw him with another woman - she has been single ever since.
She said: "You call if off if you've been two-timed, but I've still got the ring." Despite injuring her leg in a fall, Nellie has lived in the same house since 1910 in Bradford, West Yorks. "I wouldn't be anywhere else," she said.


text:
ALL STAR / Pro Leather 76 ...
Refine by
- 1
Pro Leather Tumbled Leather Low Top Deep Bordeaux$140.00
Incl. Tax: $70.00
Pro Leather Tumbled Leather Low Top Egret$140.00
Incl. Tax: $70.00
Pro Leather 76 Tumbled Leather Low Top Black$140.00
Incl. Tax: $70.00
Refine by
- 1
Please note you are shopping in $NZD mode
What currency will I be charged in?
We only charge you in Australian Dollars, no matter what currency mode you select.
Exchange rate
We use WebserviceX to fetch the latest NZ exchange rates but these rates fluctuate so the actual cost may vary.
Am I Charged GST?
We do not charge GST on NZ purchases. Price displayed in NZD are exclusive of GST.
New Zealand customers get free express delivery on orders over AUD$90.


text:
50 summer adventures: easy
Ride, hike, kayak, swim, raft, cycle: we've 50 ideas to stretch yourselves this summer. Here are those rated as an easy challenge
Stroll the Costa Brava
If this Catalan coast hike were a paperback, it would be Laurie Lee's As I Walked out One Midsummer Morning — an easily digested delight that rambles along at a leisurely pace and gets better with each outing. Starting in the pretty resort of Llafranc, the walk combines secret beaches, stupendous scenery and stylish hotels, where you'll find your luggage waiting at the end of each day. With Inntravel (01653 617906, www.inntravel.co.uk), the seven-night trip starts at £748pp, half-board, including flights from London; the truncated four-night version starts at £435pp.
A slow spin in the Sierra Nevada
If you want less blood, less sweat and more beers on your holiday, try an Easy Going week from Pure Mountains (00 34 958…


text:
|Publication number
|Publication type
|Application number
|Publication date
|May 18, 1993
|Filing date
|Feb 8, 1991
|Priority date
|Feb 20, 1990
|Fee status
|Publication number
|John J. Nestor, Jr., Natalie L. McClure, Humberto Arzeno
|Original Assignee
|Syntex (U.S.A.) Inc.
|Export Citation
|BiBTeX, EndNote, RefMan
|Patent Citations (22), Non-Patent Citations (22), Referenced by (11), Classifications (11), Legal Events (4)
|External Links: USPTO, USPTO Assignment, Espacenet
R1 -R2 -R3 -Ser-Tyr-R4 -Leu-R5 -Pro-R6(I)
This application is a continuation-in-part of application Ser. No. 07/482,428, filed Feb. 20, 1990, abandoned.
1) Field of the Invention
This invention relates to the solid phase synthesis of serine-containing polypeptides, such as LH-RH agonists and antagonists, by a temporary minimal protection procedure.
2) Description of Related Art
The decapeptide LH-RH (luteinizing hormone-releasing hormone) is a neurohormone produced in the hypothalamus, which stimulates the secretion of the pituitary hormones, LH (luteinizing hormone) and FSH (follicle-stimulating hormone), which in turn act on the gonads to stimulate the synthesis of steroid hormones. The pulsatile release of LH-RH, and the consequent release of LH and FSH, controls the reproductive cycle in mammals.
LH-RH analogs are nona- or decapeptides which are structurally related to LH-RH and exhibit biological activity similar to that of LH-RH by acting on the pituitary--gonadal axis in competition with naturally occurring LH-RH. The analogs are the subject of intensive clinical investigation due to their demonstrated ability to alleviate the symptoms of endometriosis, prostate cancer, precocious puberty, and other hormonally mediated disorders. While certain LH-RH analogs are currently available for therapeutic use, their synthesis is a complicated and, consequently, expensive procedure which necessarily increases their ultimate cost. LH-RH analogs are conventionally described as either agonists or antagonists, depending upon their mode of action.
The paradoxical ability of the agonists to suppress gonadal function at high doses has been exploited in the therapeutic arena by the development and marketing of such drugs as Synarel� nafarelin acetate nasal solution for the treatment of endometriosis. The amino acid sequence of nafarelin is as follows: ##STR1##
Nafarelin and other LH-RH agonists such as leuprorelin, buserelin, goserelin, histrelin, triptorelin and deslorelin all differ from naturally occurring LH-RH by replacement of at least the glycine residue at the 6-position with a D-amino acid. The synthetic agonists then have, in common with the naturally occurring hormone, histidine at position 2, serine at position 4, tyrosine at position 5, and arginine at position 8, all of which have reactive side chains which may present synthetic difficulties.
The LH-RH antagonists directly suppress gonadotropin secretion and are, therefore, desired for their immediate and profound effects. Generally, antagonism requires deletion or replacement of the histidyl residue at position 2 in the naturally occurring hormone. From the synthetic perspective, the deletion of histidine reduces the opportunities for undesired side reactions; however, the presence of arginine, serine and tyrosine still requires that special steps be taken to avoid side reactions.
LH-RH analogs may be synthesized by various methods, such as are taught by J. M. Stewart and J. D. Young, Solid Phase Peptide Synthesis, W. H. Freeman Co., San Francisco, 1969; J. Meinenhofer, Hormonal Proteins and Peptides, Vol. 2, page 46, Academic Press (New York), 1973; and E. Schroder and K. Lubke, The Peptides, Vol. 1, Academic Press (New York), 1965. The methods may be broadly characterized as either solution phase or solid phase techniques. Both methods involve the sequential addition of amino acids to a growing peptide chain; however, the more rigorous conditions of solid phase synthesis generally require that any reactive side chains on the amino acids be protected during formation of the amide linkage. The side chain protecting groups may be removed concurrently with the cleavage of the completed polypeptide from the inert support on which it is made or in a separate deprotection step.
One particularly useful solid phase synthetic method for preparing LH-RH analogs is disclosed in Nestor et al., U.S. Pat. No. 4,234,571. In this commonly used approach, the α-amino (N.sup.α) function of each amino acid is protected by an acid sensitive group, such as t-butoxycarbonyl (Boc); any reactive side chains, as are present on arginine, serine, histidine and tyrosine, are also protected with strongly bound groups which require treatment with hydrogen fluoride or similarly drastic procedures for their removal. This use of N.sup.α -Boc protection and HF labile side chain protection, while adequate for the preparation of research quantities of peptides, is not entirely satisfactory for large scale production. The fully protected amino acids are expensive and require a separate step for deprotection. Further, the use of hydrogen fluoride for the final deprotection, in addition to posing serious environmental hazards, contributes to commercially unacceptable yield losses.
Alternate protocols are no more appealing. Tien et al., Pept. Chem. 375-379, T. Shiba and S. Sakakibara (Ed.), Protein Research Foundation, Osaka (1988), have reported the synthesis of LH-RH using tosyl protection on histidine and benzyl protection on tyrosine and serine. This approach, while avoiding the use of hydrogen fluoride, still requires a separate dehydrogenation step to remove the benzyl protecting groups, with some reduction of tryptophan occurring.
D. H. Coy et al., Int. J. Peptide Protein Res., 14, 339-343 (1979) report the synthesis of the LH-RH antagonists, [D-Phe2,D-Trp3, D-Phe6 ]LH-RH using a variety of side chain protection protocols, all of which require HF deprotection: providing benzyl side chain protection of serine only (with strong acid salt protection of arginine), tosyl side chain protection of arginine only, both serine and arginine side chain protection, and serine, arginine and tyrosine (with 2-bromobenzyloxycarbonyl) side chain protection. N.sup.α -Boc protection was employed in all cases and removed between steps with 33% trifluoroacetic acid in methylene chloride. Hydrogen fluoride was used to cleave the crude peptide from its support and to remove the side chain protecting groups. An "unprotected" synthesis employing only salt protection of arginine (as Arg.HCl) was also conducted. All of the protected syntheses gave poorer yields than that of the unprotected side chain synthesis which did not require HF treatment.
Coy et al. further reported the synthesis of the LH-RH agonist [D-Leu6, desGly-NH2 10 ]LH-RH ethylamide with dinitrophenyl side chain protection of histidine only, salt protection of arginine, and no HF treatment. The dinitrophenyl side chain protecting group was removed during cleavage of the peptide from its support with a solution of ethylamine in dimethylformamide. The yield from the histidine protected synthesis was 34% versus 24% for a fully protected, HF cleavage synthesis. No comparison with an unprotected synthesis was made.
The art suggests that, of the various minimal protection strategies, histidine-only protection may provide some improvement in yield over fully protected syntheses for certain LH-RH agonists; however, no particular benefit is associated with any of the reported side chain protection tactics for LH-RH antagonists.
While the ideal approach for eliminating the HF deprotection step may be to conduct an unprotected synthesis, lack of protection for histidine leads to excessive racemization. Following Coy et al., however, we have found that the use of histidine-only protection results in high levels of a bis-serine impurity. Significant improvement over the teachings of the art is needed in order to obtain a practicable minimal protection synthesis for LH-RH analogs that does not require an HF deprotection step yet provides protection for those groups which, if unprotected, will adversely affect the purity and yield of peptide.
The disclosures of the aforementioned patents and publications are incorporated by reference herein.
It is an object of this invention to provide a process for the synthesis of LH-RH analogs in which the side chains of only an essential minimal number of amino acid residues are protected.
It is a further object of this invention to provide a process for the synthesis of LH-RH analogs which obviates the need for an HF deprotection step.
It is also an object of this invention to provide a process for the synthesis of serine-containing polypeptides in which formation of serine-related impurities is inhibited.
The objects of this invention are achieved for the synthesis of serine-containing polypeptides by a process in which the side chain of the amino acid residue serine is temporarily protected with a protecting group which is removed immediately following the addition of the serine to the peptide chain. The side chain protecting group is one which is labile to the same agents useful for removing the α-amino protecting group. For those peptides which contain a histidine residue, the imidazole side chain is protected with a base sensitive group.
Temporary side chain protection of serine and side chain protection of histidine, if present, minimizes formation of impurities and maximizes yields without requiring an HF or alternative separate deprotection step.
The abbreviations for the various common amino acids are those recommended by the IUPAC-IUB Commission on Biochemical Nomenclature, Biochemistry, 11, 1726 (1972). All peptide sequences mentioned herein are written according to the generally accepted convention whereby the N-terminal amino acid is on the left and the C-terminal amino acid is on the right.
The abbreviations herein represent L-amino acids, with the exception of the achiral amino acid glycine, and with the further exception of any unnatural amino acids which are achiral, or are otherwise designated as D- or D,L-. Et is ethyl, Bu is butyl, and iPr is iso-propyl.
Other abbreviations useful in describing the invention involve replacements of the amino acids in the natural LH-RH peptide by the following:
______________________________________Amino acid residue Abbreviation______________________________________3-(2-naphthyl)-alanyl Nal(2)3-(p-fluorophenyl)-alanyl p-F--Phe3-(p-chlorophenyl)-alanyl p-Cl--Phe3-(3-pyridyl)-alanyl Pal(3)NG,NG' -bis(ethyl)- hArg(Et)2homoarginylNG,NG' -bis(2,2,2- hArg(CH2 CF3)2trifluoroethyl)-homoarginylNG -butyl- hArg(Bu)homoarginylN.sup.ε -isopropyl-lysyl Lys(iPr)(benzyl)-histidyl His(Bzl)______________________________________
As used herein, the term "pharmaceutically acceptable salts" refers to salts that retain the desired biological activity of the parent compound without toxic side effects. Examples of such salts are acid addition salts formed with inorganic acids, for example hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid and the like; and salts formed with organic acids such as, for example, acetic acid, oxalic acid, tartaric acid, succinic acid, maleic acid, fumaric acid, gluconic acid, citric acid, malic acid, ascorbic acid, benzoic acid, tannic acid, pamoic acid, alginic acid, polyglutamic acid, naphthalenesulfonic acids, naphthalenedisulfonic acids, polygalacturonic acid, and the like.
Temporary protection means that the serine side chain is protected for a relatively short period of the synthetic cycle. The side chain protecting group and the α-amino protecting group are removed simultaneously, after the serine coupling is effected. Generally, the critical criterion for selecting the serine side chain protecting group is that the group be stable to coupling conditions but labile to α-amino deprotecting conditions. The serine side chain is preferably protected by a group selected from t-butyl, t-butyldimethylsilyl, trimethylsilyl, trityl, pivalyl, and tetrahydropyran-2-yl.
For those peptides which have histidine residues it is generally desirable to protect the imidazole side chain in order to avoid racemization during coupling. This protection may also be of the temporary variety, i.e. labile during the coupling cycle, or may remain in place until the peptide is removed from its support. Preferably, ammonolysis is used to cleave the peptide from its support and simultaneously remove the histidine protecting group. Preferred side chain protecting groups for histidine are p-toluenesulfonyl and 2,4-dinitrophenyl.
In another aspect of this invention, the deprotecting agent is selected from solutions of hydrogen chloride in C3 to C6 alcohols and dichloromethane. Preferably, the ratio of alcohol to dichloromethane is from 0.1 to 10.0 (v/v) and the acid concentration is 2N to 9N. Most preferably, the alcohol is i-propanol.
The temporary minimal protection process of this invention is expected to be applicable to the solid phase synthesis of any serine-containing polypeptide having a few to several dozen residues, regardless of the remainder of the sequence. LH-RH analogs, and other nona- and decapeptides, are preferred synthetic targets. While the invention is described with reference to the sequential addition of individual amino acids, those skilled in the art will recognize that the process is equally applicable to syntheses in which blocks of smaller polypeptides are coupled to form a larger polypeptide, e.g. by adding a tetrapeptide to a pentapeptide, provided that the side chains of any serine residues are temporarily protected during the serine coupling cycle.
In one embodiment of the invention there is provided an improved minimal protection process for the solid-phase synthesis of a compound having an amino acid sequence of the formula
R1 -R2 -R3 -Ser-Tyr-R4 -Leu-R5 -Pro-R6 (I)
R1 is selected from (pyro)Glu and N-Ac-D-Nal(2);
R2 is selected from His, D-p-Cl-Phe and D-p-F-Phe;
R3 is selected from Trp, D-Trp, D-Nal(2) and D-Pal(3);
R4 is selected from D-Nal(2), D-hArg(Et)2, D-Lys(iPr), D-hArg(Bu), D-hArg(CH2 CF3)2, D-His(Bzl), D-Leu, D-Pal(3), D-Ser(tBu) and D-TGrp;
R5 is selected from Arg, hArg(Et2), hArg(Bu), hArg(CH2 CF3)2 and Lys(iPr); and
R6 is selected from Gly-NH2, NH--NHCONH2, D-Ala-NH2 and NHEt;
wherein the amino acids are provided with N.sup.α protection;
in which the improvement comprises (a) temporarily protecting the side chain of serine at position 4 and (b) protecting the side chain of histidine, if present, with a base sensitive group.
Preferably, the side chain of serine at position 4 is protected with a group labile to those agents useful for removing N.sup.α protecting groups without inducing racemization, side reactions, or cleavage of the growing peptide from its resin support.
In a preferred embodiment, there is provided a process for the production of a polypeptide having the formula above wherein,
R1 is (pyro)Glu or N-Ac-D-Nal(2);
R2 is His or D-p-Cl-Phe;
R3 is Trp or D-Pal(3);
R4 is D-Nal(2), D-Leu, D-Trp, D-Ser(tBu), D-His(Bzl) or D-hArg(Et)2 ;
R5 is Arg or hArg(Et)2 ; and
R6 is Gly-NH2, NHEt or D-Ala-NH2.
Most preferably, the invention provides a process for the production of nafarelin, on a styrene-divinylbenzene resin, using N.sup.α -Boc protection, wherein,
R1 is (pyro)Glu,
R2 is His,
R3 is Trp,
R4 is D-Nal(2)
R5 is Arg, and
R6 is Gly-NH2.
In the preferred embodiment the α-amino (N.sup.α) function of the amino acids is protected by an acid or base sensitive group. The protecting group is stable to the conditions of peptide bond formation, while being readily removable without destruction of the growing peptide chain or racemization of any of the chiral centers contained therein. Suitable N.sup.α protecting groups are t-butoxycarbonyl (Boc), biphenylisopropyloxycarbonyl, T-amyloxycarbonyl, isobornyloxycarbonyl, α,α-dimethyl-3,5-dimethoxybenzyloxycarbonyl, o-nitrophenylsulfenyl, 2-cyano-t-butyloxycarbonyl, 9-fluorenylmethyloxycarbonyl (Fmoc) and the like. Preferably the α-amino protecting group is t-butoxycarbonyl (Boc).
The hydroxy side chain of the serine residue is protected during the coupling of serine to the growing peptide, as described for the generic embodiment of this invention. The side chain protecting group is removed after the coupling is effected and prior to adding the next amino acid. The serine side chain protecting group is removed with the same agent used to remove the N.sup.α protecting group. Preferred side chain protecting groups for serine are t-butyl, t-butyldimethylsilyl, trimethylsilyl, trityl, pivalyl and tetrahydropyran-2-yl.
Further, the imidazole side chain of histidine, generally present in LH-RH agonists, is protected. The histidine side chain protecting group is base sensitive, and may be removed during the coupling cycle, but, optionally, its removal may be completed when the peptide is cleaved from its support. Preferred side chain protecting groups for histidine are p-toluenesulfonyl and 2,4-dinitrophenyl.
To initiate the synthesis the first amino acid, which will be the C-terminal amino acid in the final product, is attached to a suitable solid support. Suitable solid supports useful for the above synthesis are those materials which are inert to the reagents and reaction conditions of the stepwise condensation-deprotection reactions, as well as being insoluble in the media used. Examples of commercially available resins include styrene/divinylbenzene resins modified with a reactive group, e.g., chloromethylated styrene/divinylbenzene copolymer, hydroxymethylated styrene-divinylbenzene copolymer, and the like. Merrifield resin (1% crosslinked chloromethylated styrene/divinylbenzene copolymer) is preferred.
The attachment to the chloromethylated styrene divinylbenzene resin is made by means of the reaction of the N.sup.α protected C-terminal amino acid, especially the N.sup.α -Boc amino acid, as its cesium, tetramethylammonium, triethylammonium, 1,5-diazabicyclo [5.4.0]undec-5-ene, or similar salt in ethanol, acetonitrile, N,N-dimethylformamide (DMF), and the like, especially the cesium salt in DMF, with the chloromethylated resin at an elevated temperature, for example between about 40� and 60� C., preferably about 50� C., for from about 12 to 72 hours, preferably about 48 hours.
The successive coupling of the protected amino acids is carried out by methods well known in the art, typically in an automated polypeptide synthesizer. Each protected amino acid is introduced in from about 1.5 to about 2.5 fold molar excess and the coupling is carried out in an inert, nonaqueous, polar solvent such as dichloromethane, DMF or mixtures thereof, preferably in dichloromethane at about ambient temperature. The coupling agent is selected from N,N'-dicyclohexylcarbodiimide (DCC), N,N'-diisopropylcarbodiimide (DIC) or other carbodiimide either along or in the presence of 1-hydroxybenzotriazole (HBt), O-acyl ureas, benzotriazole-Δ-yl-oxy-tris(pyrrolidino phosphonium) hexafluorophosphate (PyBop), N-hydroxysuccinimide, other N-hydroxyimides or oximes. Alternately, protected amino acid active esters (e.g., p-nitrophenyl, pentafluorophenyl and the like) or symmetrical anhydrides may be used.
The peptide resin is checked for complete coupling using the Kaiser Test (Anal. Biochem., 34, 595 (1970)), except for the coupling to proline in which case the Chloranil Test (Anal. Biochem., 117, 145 (1981)) or the Isatin Test (Anal. Chim. Acta, 118, 149 (1980)) is used.
If the completion test(s) suggest that the reaction is not complete, the coupling is repeated using additional amino acid but omitting additional acid deprotection. When the last coupling is completed, the resin is washed with methanol or methanol containing dichloromethane and dried at a maximum of 60� C.
At the end of each cycle, i.e., after each successive N.sup.α -protected amino acid is added to the growing polypeptide chain, the protecting group is removed by treatment with a deprotecting agent. After serine is incorporated, the deprotecting agent removes both the N.sup.α -Boc protecting group and the hydroxy side chain protecting group. The preferred deprotecting agents include hydrogen chloride in dichloromethane (HCl/CH2 Cl2) and hydrogen chloride dissolved in a C3 -C6 alcohol, preferably isopropanol, mixed with dichloromethane. Generally, the concentration of the HCl will be 2N to 9N, preferably 4N to 5N. The ratio of CH2 Cl2 to the C3 -C6 alcohol is 0.1 to 10 (v/v), preferably about 1:1. A particularly preferred deprotecting agent is 4.5N HCl in i-PrOH:CH2 Cl2 (1:1). The deprotection step generally takes place at temperatures of 0� C. to 45� C., preferably at ambient temperatures (20� to 27� C.).
Those skilled in the art will appreciate that selection of a coupling/deprotection protocol utilizing agents other than those described above is entirely appropriate provided that the serine residue is deprotected with an agent which accomplishes the objectives of this invention. A protocol which uses HCl/iPrOH/CH2 Cl2 for each deprotecting cycle may be employed. Alternatively, a mixed protocol in which TFA/CH2 Cl2 is used for certain cycles and HCl/iPrOH/CH2 Cl2 for others is also useful. Other cycles will be readily apparent to the skilled artisan.
When it is desired to prepare a peptide, such as buserelin or goserelin, in which R4 in Formula (I) above is D-Ser(tBu), a base sensitive N.sup.α protecting agent, such as Fmoc, is required for N.sup.α protection. Fmoc is labile to basic agents (pH>8.5), such as piperidine, which will not remove the tBu from the D-Ser(tBu). In later cycles, following the addition of the D-Ser(tBu), it is also necessary to use base sensitive N.sup.α protection. The side chain of the serine at position 4 is protected with a group which is removed with an agent which will not remove the tBu from D-Ser (t Bu), such as a mild fluoride treatment (1 equiv.) or ammonolysis. A preferred serine side chain protecting group of this embodiment is t-butyldimethylsilyl. In this manner, a D-Ser (tBu) residue may be present in the final peptide, while practicing temporary protection of the serine at position 4.
At the end of the solid phase synthesis the polypeptide is cleaved from the resin. For peptides with an alanine or glycine C-terminus, cleavage is by ammonolysis with a saturated solution of ammonia in a suitable solvent; for those peptides having a proline C-terminus, cleavage is by aminolysis with an alkylamine or fluoroalkylamine. For peptides with an alanine or glycine C-terminus, the cleavage is conducted at temperatures between about 10� and 50� C., preferably about 25� C., for between about 12 and 24 hours, preferably about 18 hours. Suitable solvents include methanol, ethanol, isopropanol, dimethylformamide, tetrahydrofuran, N,N-dimethylethanolamine, hexanes and mixtures thereof. Preferably, a saturated solution of ammonia in methanol is used. Alternatively, the peptide may be removed from the resin by transesterification with a base, followed by aminolysis.
The polypeptide is then purified by a sequence of chromatographic steps employing any or all of the following types: ion exchange on a weakly basic resin in the acetate form; hydrophobic adsorption chromatography on underivatized polystyrene-divinylbenzene (e.g., Amberlite� XAD); silica gel adsorption chromatography; ion exchange chromatography on carboxymethylcellulose; partition chromatography (e.g., on Sephadex� G-25), or countercurrent distribution; high performance liquid chromatography (HPLC), especially reversed-phase HPLC on octyl- or octadecyl-silylsilica bonded phase column packing.
If a racemic amino acid is used in one or more of the 1, 2, 3 or 6 positions and individual isomeric products are desired, the diastereomeric nonapeptide or decapeptide final products are separated, and the desired peptide containing a D-amino acid in the appropriate position is isolated and purified, preferably during the above-described chromatographic process.
Optionally, the isolated and purified polypeptide is converted to a pharmaceutically acceptable salt.
The following Examples compare the temporary protection process of this invention with an unprotected process for both an LH-RH agonist and an LH-RH antagonists. These Examples are presented for purposes of specificity only and should not be construed so as to place any undue limitations on the scope of the claimed invention.
N.sup.α -Boc amino acids were obtained from Bachem (Torrance, Calif.) (Leu, Tyr, His(Tos), Arg, Trp, D-Ala and Gly), Star Biochemicals (Torrance, Calif.) (Pro and Ser(tBu)), Synthe Tech (Albany, Ore.) (D-Nal(2)), Incell (Milwaukee, Wis.) (D-Pal(3) and UCB Bioproducts (Belgium) (p-Cl-Phe).
Solutions of 4-4.5N HCl in i-PrOH/CH2 Cl2 (1/1) were prepared by bubbling HCl into cooled i-PrOH. Once the solution became saturated (determined by titration, approximately 9N), the solution was kept at room temperature for no more than 3 days and diluted with an equal volume of CH2 Cl2 before use.
The following protocols were used to remove the N.sup.α -protecting group following each addition.
Program A: The resin was first washed with CH2 Cl2 1�1 min., TFA-CH2 Cl2 (40/60) 1�1 min., TFA-CH2 Cl2 (40/60) 1�30 min., CH2 Cl2 5�1 min., Et3 N-CH2 Cl2 (5/95) 3�1 min., CH2 Cl2 4�1 min.
Program B: The resin was first washed with CH2 Cl2 1�1 min., 4-4.5N HCl in CH2 Cl2 /i-PrOH (1/1) 1�1 min., 4-4.5N HCl in CH2 Cl2 /i-PrOH (1/1) 1�30 min., CH2 Cl2 3�1 min., DMF 1�1 min., Et3 N-CH2 Cl2 (5/95) 3�1 min., DMF 1�1 min., CH2 Cl2 4�1 min.
After each deprotecting and washing step, following protocol A or B, the next amino acid in sequence was added and the resin washed with CH2 Cl2 3�1 min., MeOH 4�1 min., DMF 2�1 min. and CH2 Cl2 4�1 min.
When the sequence was completed, the peptide was cleaved from the resin by treatment with a saturated solution of ammonia in methanol for about 18 hours at about 25� C.
The yields obtained in both Examples 1 and 2 using the temporary minimal protection process of this invention are significantly increased over the yields obtained in Comparative Examples 1 and 2 using unprotected syntheses.
In addition to the increase in yield, the process of this invention offers the additional advantages of employing less hazardous reagents, over a shorter time period and with lower energy expenditures in the isolation and purification of LH-RH analogs. A further advantage is the generation of smaller amounts of a considerably less toxic waste stream.
4.9 g of N.sup.α -Boc-glycine was dissolved in a mixture of 50 ml. methanol and 50 ml. distilled water. The pH of the solution was brought to 7.5 with aqueous cesium bicarbonate. The solvent was then removed under vacuum.
After 18 hours of drying under high vacuum, the residue was dissolved in 150 ml. dry DMF. 25 g 1% chloromethylated polystyrene/divinylbenzene (Merrifield) resin (corresponding to 25 mmole chloride) was added. The mixture was shaken at 50� C. for 24 hours, filtered, and the resin was then washed sequentially with DMF, water, and ethanol. The resin was dried under vacuum for 3 days to yield 28.34 g of Boc-Gly-O-Resin.
Following the procedures of Preparation A, N.sup.α -Boc-D-alanine was added to 1% Merrifield resin to provide N.sup.α -Boc-D-Ala-O-resin.
In this Example, nafarelin was prepared using the following side chain protection protocol: salt protection for arginine (as the chloride), tosyl protection for histidine His(Tos), and t-butyl protection for serine Ser(tBu).
1.0 mmol of N.sup.α -Boc-Gly-O-resin from Preparation A was placed in the reaction vessel of a 5.0 L Vega 296 automated solid phase peptide synthesizer fitted with accessory bottles and flasks for addition of reagents and for pressurization, depressurization and maintenance of an inert atmosphere of nitrogen.
The following amino acids were added to the N.sup.α -Boc-Gly-O-resin by DIC or HBt-assisted DIC coupling for 3 hours:
N.sup.α -Boc-Pro 2.0 equiv.
N.sup.α -Boc-Arg.HCl 2.0 equiv.
N.sup.α -Boc-Leu.H2 O 2.0 equiv.
N.sup.α -Boc-D-Nal(2) 1.5 equiv./HBt
N.sup.α -Boc-Tyr 1.5 equiv./HBt
N.sup.α -Boc-Ser(tBu) 2.0 equiv./HBt
N.sup.α -Boc-Trp 1.75 equiv./HBt
N.sup.α -Boc-His(Tos 1.75 equiv./HBt
(pyro)Glu 2.5 equiv./HBt
Program A was used to remove the N.sup.α protecting groups on Gly, Pro, Arg, Leu, D-Nal(2) and Tyr. Program B was used for the removal of the N.sup.α protecting groups on Ser, Trp, and His and for the removal of the serine side chain protecting group.
The crude peptide was dissolved in 2M acetic acid and converted to the acetate salt by passage through a column of AG3-X4A resin (Bio-Rad). The acetate was dissolved in a minimal amount of methanol and acetone added to reprecipitate the peptide. Reversed phase HPLC (Partisil ODS-3, 40μ, acetonitrile with 0.5% acetic acid) was used to remove polar and non-polar impurities. Fractions containing at least 97% nafarelin acetate were combined, diluted with water, reloaded on a reversed-phase HPLC column, and washed with 1% acetic acid in water. The residue was precipitated, filtered, washed with ether and dried under vacuum.
Amino acid analyses were performed on a Beckman 119CL amino acid analyzer. Samples for amino acid analyses were hydrolyzed with 4N CH3 SO3 H (0.2% 3-(2-aminomethyl indole) HCl) for 20 hrs at 110� C.
Analytical HPLC was performed on a Spectra Physics 8800 chromatograph, using an ODS-II column from Alltech, 5μ, 4.6�250 mm, 10 μl inj., flow - 1.5 ml/min., 27.5% CH3 CN, 72.5% 0.16M KH2 PO4 pH=5.1, temp.=40� C.
HPLC analysis of the crude peptide showed a main peak with a retention time (rt) of 18 min. corresponding to nafarelin and no impurity over 1% at rt 14 min.
The procedure of Example 1 was followed except that N.sup.α -Boc-Ser was substituted for N.sup.α -Boc-Ser(tBu).
HPLC analysis showed a main peak at 18 min. corresponding to nafarelin and 8.1 to 11.5% of an impurity at a retention time (rt) of 14 min.
The yield from this "unprotected" synthesis was approximately the same as that obtained from a fully protected synthesis and significantly lower than that achieved with the temporary protection synthesis of Example 1.
In this Example an LH-RH antagonist, ganirelix, N-Ac-Nal(2)-D-pCl-Phe-D-Pal(3)-Ser-Tyr-D-hArg(Et)2 -Leu-hArg(Et)2 -Pro-Ala-NH2, was prepared using the following side chain protection protocol: salt protection for L- and D-hArg(Et)2 (as the chloride) and t-butyl protection for serine.
Amino acids were added to the N.sup.α -Boc-D-Ala-O-Resin of Preparation B in the following sequence:
N.sup.α -Boc-Pro 2.3 equiv.
N.sup.α -Boc-hArg(Et)2.HCl 1 equiv./HBt
N.sup.α -Boc-Leu.H2 O 2.3 equiv.
N.sup.α -Boc-D-hArg(Et)2.HCl 1.6 equiv./HBt
N.sup.α -Boc-Tyr 2.1 equiv./HBt
N.sup.α -Boc-Ser(tBu) 2.0 equiv.
N.sup.α -Boc-D-Pal(3) 1.8 equiv./HBt
N.sup.α -Boc-D-p-Cl-Phe 2.0 equiv.
N.sup.α -Boc-D-Nal(2) 2.1 equiv./HBt
Acetic anhydride
An acetylation (capping of the resin) was done after Ala, Pro and Leu. Excess HBt (2 equiv.) was used for the coupling of the basic amino acids, hArg(Et)2 and Pal(3).
Program A was used for the removal of the protecting groups on Ala, Pro, L-hArg(Et)2, Leu and D-Nal(2); Program B was used for the removal of the protecting groups on D-hArg(Et)2, Tyr, Ser, D-Pal(3) and p-Cl-Phe.
The crude peptide was first dissolved in 2M acetic acid and converted to its acetate salt by passage through a column of AG3-X4A resin (Bio-Rad). The acetate was subjected to chromatography on a silica gel column (Cl2 /i-PrOH/MeOH/H2 O/HOAc solvent); the acetate fractions dissolved in H2 O and loaded onto a reversed-phase column (Vydec C-18, 15-20μ), and purified using acetonitrile/TEAP (pH 3). Fractions of the desired purity were combined and diluted with water and reloaded on a reversed-phase HPLC column, then washed with 1% acetic acid in water. The peptide was stripped with a mixture of MeOH/CH3 CN/HOAc/H2 O (44/50/1/5). The residue was dissolved in acetic acid and precipitated over ether, filtered, washed with ether and dried under vacuum.
Amino acid analyses were performed on a Beckman 119CL amino acid analyzer. Samples for amino acid analyses were hydrolyzed with 6N HCl at 110� C. for 20 hrs.
Analytical HPLC was performed on a Spectra Physics 8800 chromatograph, using a Spherisorb C-8 (Alltech), 5μ, 4.6�250 mm, 10 μl inj., flow=1.5 ml/min., 30% CH3 CN, 70% NH4 H2 PO4 0.04M, dimethyloctylamine 4.3�10-3, temp. 40� C.
Snythesis of ganirelix was confirmed by the presence of a main peak at rt 18 min.; no other peak over 1% was noted at rt 16 min.
Example 2 was repeated using N.sup.α -Box-Ser instead of N.sup.α -Boc-Ser(tBu).
HPLC analysis showed the presence of a main peak at 18 min. corresponding to ganirelix and the presence of an impurity in 6.5% at rt 16 min.
The following claims particularly point out and distinctly claim the subject matter which applicants regard as their invention. These claims are entitled to the full range of equivalents recognizable by those skilled in the art of solid phase peptide synthesis.
|Cited Patent
|Filing date
|Publication date
|Sep 13, 1971
|Jul 30, 1974
|Abbott Lab
|Protected decapeptide derivatives of gonadotropin releasing hormone
|Feb 22, 1973
|Oct 15, 1974
|American Home Prod
|Homo-(tyr5')-lrf and thr4-lrf and intermediates
|Oct 3, 1973
|Dec 17, 1974
|American Home Prod
|P-glu-d-phe-t rp-ser-tyr-d-ala-leu-arg-pro-gly-nh2-and intermediates
|Jul 5, 1974
|Jun 17, 1975
|American Home Prod
|Method of inhibiting fertility in mammals using D{13 Phe{hu 2{b {13 D{13 Ala{hu 6{b {13 LRF
|Mar 13, 1974
|Aug 26, 1975
|American Home Prod
|P-glu-his-trp-ser-tyr-d-pgl-leu-arg-pro-gly-nh' 2 'and intermediates
|Nov 4, 1974
|Dec 23, 1975
|American Home Prod
|P-Glu-D-p-F-Phe-Trp-Ser-Tyr-D-Ala-Leu-Arg-Pro-Gly-NH{HD 2
|Jan 27, 1975
|Jan 6, 1976
|American Home Products Corporation
|(H-gly-gly-tyr-ala)1 -somatostatin
|Nov 14, 1974
|Jan 20, 1976
|American Home Products Corporation
|(N-Acetyl)-Pro-D-Phe-Trp-Ser-Tyr-D-Ala-Leu-Arg-Pro-NHEt and intermediates
|Mar 28, 1975
|Mar 2, 1976
|American Home Products Corporation
|PGlu-D-Met-Trp-Ser-Tyr-D-Ala-Leu-Arg-Pro-Gly-NH2 and intermediates
|Feb 1, 1977
|Aug 22, 1978
|Hoffmann-La Roche Inc.
|Solid phase synthesis with base N alpha-protecting group cleavage
|Jun 11, 1979
|Nov 18, 1980
|Syntex (U.S.A.) Inc.
|Nonapeptide and decapeptide derivatives of luteinizing hormone releasing hormone
|May 8, 1980
|Nov 17, 1981
|American Home Products Corp.
|Preparation of (D-Trp 6)-LH-RH via the heptapeptide H-Ser-Tyr-D-Trp-Leu-Arg-Pro-Gly-NH2
|Jun 2, 1980
|Mar 2, 1982
|Coy David Howard
|LH-RH Antagonists
|Jan 20, 1982
|Feb 14, 1984
|Coy David Howard
|LH-RH Antagonists
|Dec 21, 1982
|Nov 6, 1984
|Syntex (U.S.A.) Inc.
|Nonapeptide and decapeptide analogs of LHRH useful as LHRH antagonists
|Apr 26, 1985
|Apr 7, 1987
|Board Of Regents, The University Of Texas System
|Effective hormonal peptides: D-3-QA1 6-LHRH
|May 20, 1983
|May 19, 1987
|Syntex (U.S.A.) Inc.
|Nonapeptide and decapeptide analogs of LHRH, useful as LHRH antagonists
|Dec 23, 1986
|Apr 26, 1988
|The Salk Institute For Biological Studies
|GnRH antagonists VIII
|Feb 5, 1987
|Jan 31, 1989
|Syntex (U.S.A.) Inc.
|Nonapeptide and decapeptide analogs of LHRH useful as LHRH antagonists
|Apr 9, 1986
|Aug 8, 1989
|Alpha-1 Biomedicals, Inc.
|Solid phase process for synthesizing peptides
|Mar 21, 1988
|Aug 22, 1989
|Ciba-Geigy Corporation
|Synthetic polystyrene resin and its use in solid phase peptide synthesis
|Aug 24, 1987
|Jun 19, 1990
|Board Of Regents, The University Of Texas System
|Effective antagonists of the luteinizing hormone releasing hormone which release negligible histamine
|Coy et al. Int. J. Prot. Res. vol. 14 339 343 (1979).
|Coy et al. Int. J. Prot. Res. vol. 14 339-343 (1979).
|D. H. Coy, et al., Minimal Side Chain Protection Can Be A Successful Strategy in Solid Phase Peptide Synthesis, Int. J. Peptide Protein Res., 14, 339 343 (1979).
|D. H. Coy, et al., Minimal Side-Chain Protection Can Be A Successful Strategy in Solid-Phase Peptide Synthesis, Int. J. Peptide Protein Res., 14, 339-343 (1979).
|Hocart et al. CA 106: 120222c (1987), J. Med. Chem (1987) 30(4) 739 43.
|Hocart et al. CA 106: 120222c (1987), J. Med. Chem (1987) 30(4) 739-43.
|Hocart et al. CA 107:134666h (1987), J. Med. Chem (1987) 30(10) 1910 14.
|Hocart et al. CA 107:134666h (1987), J. Med. Chem (1987) 30(10) 1910-14.
|Houghten et al. Int l. J. Pept. Prot. Res. vol. 27 (1986) 653 58.
|Houghten et al. Int'l. J. Pept. Prot. Res. vol. 27 (1986) 653-58.
|J. K. Inman, Peptide Synthesis with Minimal Protection of Side Chain Functions, The Peptides, 3, 253 302 (Academic Press, 1981).
|J. K. Inman, Peptide Synthesis with Minimal Protection of Side-Chain Functions, The Peptides, 3, 253-302 (Academic Press, 1981).
|J. Tien, et al., Novel Process for the Production of LH RH Using CTH and Preparative HPLC, Peptide Chemistry 1987, 375 377 (Protein Research Foundation, Osaka, 1988).
|J. Tien, et al., Novel Process for the Production of LH-RH Using CTH and Preparative HPLC, Peptide Chemistry 1987, 375-377 (Protein Research Foundation, Osaka, 1988).
|K. M. Sivanandaiah, et al., Improved Solid Phase Synthesis of Luteinizing Hormone Releasing Hormone Analogues Using 9 Fluorenylmethyloxycarbonyl Amino Acid Active Esters and Catalytic Transfer Hydrogenation with Minimal Side Chain Protection and Their Biological Activities, J. Biosci., 14:3, 311 317 (1989).
|K. M. Sivanandaiah, et al., Improved Solid Phase Synthesis of Luteinizing Hormone Releasing Hormone Analogues Using 9-Fluorenylmethyloxycarbonyl Amino Acid Active Esters and Catalytic Transfer Hydrogenation with Minimal Side-Chain Protection and Their Biological Activities, J. Biosci., 14:3, 311-317 (1989).
|M. Bodanszky, et al., Side Reactions in Peptide Synthesis. 4. Extensive O Acylation by Active Esters in Histidine Containing Peptides 1 , J. Org. Chem., 42:1, 149 152 (1977).
|M. Bodanszky, et al., Side Reactions in Peptide Synthesis. 4. Extensive O-Acylation by Active Esters in Histidine Containing Peptides1, J. Org. Chem., 42:1, 149-152 (1977).
|M. Fujino, et al., Some Analogs of Luteinizing Hormone Releasing Hormone (LH RH) Having Intense Ovulation Inducing Activity, Biochem. Biophys. Res. Comm., 57:4, 1248 1256 (1974).
|M. Fujino, et al., Some Analogs of Luteinizing Hormone Releasing Hormone (LH-RH) Having Intense Ovulation-Inducing Activity, Biochem. Biophys. Res. Comm., 57:4, 1248-1256 (1974).
|R. A. Houghten, et al., Use of 10% Sulfuric Acid/Dioxane for Removal of N Tertiary Butyloxycarbonyl Group During Solid Phase Peptide Synthesis, Int. J. Peptide Protein Res., 27, 653 658 (1986).
|R. A. Houghten, et al., Use of 10% Sulfuric Acid/Dioxane for Removal of N-α-Tertiary-Butyloxycarbonyl Group During Solid Phase Peptide Synthesis, Int. J. Peptide Protein Res., 27, 653-658 (1986).
|Citing Patent
|Filing date
|Publication date
|Apr 28, 1994
|Jun 2, 1998
|United Biomedical, Inc.
|Immunogenic LHRH peptide constructs and synthetic universal immune stimulators for vaccines
|Mar 8, 2005
|Aug 4, 2009
|Ajinomoto Co., Inc.
|Method of preparing peptide
|May 21, 2009
|Mar 29, 2011
|Ajinomoto Co., Inc.
|Method of preparing peptide
|Jun 6, 2012
|Apr 11, 2017
|Polyphor Ag
|Beta-hairpin peptidomimetics
|May 21, 2009
|Sep 17, 2009
|Ajinomoto Co., Inc.
|Method of preparing peptide
|Jun 6, 2012
|Aug 6, 2015
|Polyphor Ag
|Beta-hairpin peptidomimetics
|Feb 9, 2012
|Jul 18, 2012
|Preparation method for ganirelix acetate
|Jun 16, 2014
|Sep 3, 2014
|Method for preparing ganirelix acetate
|Mar 7, 2005
|Oct 12, 2005
|Ajinomoto Co., Inc.
|Method of preparing tripeptide intermediates for the preparation of LHRH antagonists
|Oct 12, 2007
|Apr 17, 2008
|Dong Kook Pharm. Co., Ltd
|A method for preparing peptides using by solid phase synthesis
|Jun 12, 2015
|Dec 17, 2015
|Ganirelix precursor and method for preparing ganirelix acetate by using anirelix precursor
|U.S. Classification
|530/334, 530/335, 530/328, 530/313
|International Classification
|Cooperative Classification
|European Classification
|Feb 8, 1991
Owner name: SYNTEX (U.S.A.) INC., 3401 HILLVIEW AVENUE, P.O. B
Free format text: ASSIGNMENT OF ASSIGNORS INTEREST.;ASSIGNORS:NESTOR, JOHN J. JR.;MC CLURE, NATALIE L.;ARZENO, HUMBERTO;REEL/FRAME:005632/0501
Effective date: 19910208
|Sep 9, 1996
|Fee payment
Year of fee payment: 4
|Sep 28, 2000
|Fee payment
Year of fee payment: 8
|Sep 29, 2004
|Fee payment
Year of fee payment: 12


text:
Hawaiian EDventure offers an array of academic, cultural, recreational and experiential travel programs that blend Hawaii's culture, science, history and ethnic diversity. EDventure's custom-designed programs may be created to meet your educational and recreational objectives.
Our "Astronomy" program may include one, two or a three-island exploration. If you let us know what your educational/recreational needs are; Hawaiian EDventure will design the PERFECT program for your group. Below, see some sample descriptions of the Astronomy EDventure highlights.
Hawai'i Island: Stargazing on top of Mauna Kea
Mauna Kea Onizuka visitor Center: Located at 9,300 foot (2,800 m), you will receive health, safety, universal, cultural, scientific and environmental information which will permit you to understand the significance of the "White Mountain" (Mauna Kea), and get used to the altitude before reaching the summit at 13,786 foot (4,205 m). Moreover, telescopes will be set up at night to discover and see the wonder of the Universe with a passionate group of experts.
Mauna Kea observatories: Mauna Kea is one of the best locations in the World for stargazing. Its high altitude -13,786 ' (4205 m) and Hawaiian location allows around 300 clear nights per year. Therefore, breakthrough discoveries are made everyday. No wonder why Hawai'i is worlwidely famous for astronomy.
'Imiloa Astronomy Center: A unique Hawaiian voyage through Time and Space
'Imiloa Astronomy Center of Hawaii tells dual stories of the renowned Maunakea volcano, with its world-famous astronomy and rich traditions of Hawaiian culture. 'Imiloa, which means "exploring new knowledge," reflects both the Hawaiian voyages of discovery and the explorations of astronomy.