Looking for Crusher? Find out information about Crusher. machine used to reduce materials such as ore, coal, stone, ... (coal, limestones, and bauxites).
Any additional comments, directions or information the Crushers may need. Send donation requests to: Lake Erie Crushers Attn: Donation Requests 2009 Baseball Blvd.
About MCC. Shibang Industry & Technology Group Co., Ltd. is one high-tech enterprise, which involves R&D, production, sales and service as well.
Bauxite, the primary process of bauxite. Raw bauxites first enter jaw crusher to be crushed to smaller size. Chat Online; bauxite mining indo.


To filter salaries for Research Associate in Philadelphia, PA Area, Sign In or Register.
How much does a Research Associate make in Philadelphia, PA?
The average salary for a Research Associate is $53,787 in Philadelphia, PA. Salaries estimates are based on 654 salaries submitted anonymously to Glassdoor by Research Associate employees in Philadelphia, PA.


A frog goes into a bank and approaches the teller. He can see from her nameplate that her name is Patricia Whack. "Miss Whack, I'd like to get a loan to take a holiday."
Patty looks at the frog in disbelief and asks his name. The frog says his name is Kermit Jagger, his dad is Mick Jagger, and that it's okay, he knows the bank manager. Patty explains that he will need to secure the loan with some collateral.
Very confused, Patty explains that she'll have to consult with the bank manager and disappears into a back office. She finds the manager and says, "There's a frog called Kermit Jagger out there who claims to know you and wants to borrow money, and he wants to use this as collateral."
"It's a knickknack, Patty Whack. Give the frog a loan. His old man's a Rolling Stone"


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This is The End of Atheism – Debunked (a follow on from my pervious video "Questions No Atheist Can Answer - Debunked").
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In this video I played 4 songs in reverse. First we'll listen backwards, then the original. Can you recognize them?
Take Off Tokyo Haneda B777 BA Club World - Views of Skytree - Duration: 3 minutes, 15 seconds.
Take Off from Tokyo Haneda in a B777 in BA Club World rear facing window seat. Views of Tokyo Skytree prior to takeoff roll.
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Onboard a JAL 787 landing at Tokyo Haneda airport with view of an American Airlines simultaneous landing on a parallel runway.
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Hipac is the pre-recorded tape format that history forgot, so it's time to fix that.
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That Time Mozart Pirated a Forbidden Piece of Music from the Catholic Church from Memory - Duration: 9 minutes, 37 seconds.
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Brünnchen Nordschleife Hard crash bmw e36 and action highlights of touristenfahrten 14.10.2017 Nurburgring Nürburgring accident unfall.
► Nordschleife Big CRASH & FAIL Compilation 2017 !
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Chanson en allemand, en français et en alsacien avec annonce finale en alsacien diffusée sur Radio Verte Fessenheim en 1978.
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Amazing Japanese dovetail joint. Japanese joinery. - Duration: 96 seconds.
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Welcome onboard the British Airways A380 on the upper deck and let's embark on this wonderful journey from Hong Kong to London Heathrow!
BladeLess Fan - How to Make it - Dyson Fan DIY - Duration: 4 minutes, 53 seconds.
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The annual St Paul's Cathedral School's Bonfire Night Fireworks display, 16th November 2014.
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77 Italo Disco Hits In 10 Minutes!!!
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Landing at London Heathrow, runway 09L. Filmed from a British Airways Club World rear facing seat in a Boing 777.
Cockpitvideo: Crossing traffic in the sky - Duration: 17 seconds.
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Fernando Sor Op. 60 No. 13 - Duration: 37 seconds.
I like to play it faster. Sorry about the poor video quality.


True to its mission of "creating amazing" and inspired by the craftsmanship that goes into every vehicle it makes, Lexus commissioned a team of ice sculpture specialists to explore how ice could be used in place of conventional alloy metal and rubber.
Just as the skills of Lexus's famous takumi craftspeople are used to achieve the highest quality, the human hand and eye were fundamental to the success of the project. Four sculptors worked for 36 hours to produce each wheel. The millimetre-perfect finish was achieved with hand-crafting, using Japanese steel chisels and shaping tools. Even the tread patterns on the tyres were faithfully reproduced.


text:
Q: Re open the review and converter to Swift 2.0-Xcode 7 I open my project (swift 1.2) by Xcode 7, and the Xcode suggestion to me convert to Swift 2.0. But I canceled it, now I want to open the review and converter of Xcode 7. How to do that?

A: This is the answer you are looking for To Latest Swift Syntax at the latest menu.

redpajama_set_name: RedPajamaStackExchange


text:
package org.elasticsearch.xpack.ml.rest.job;

import org.elasticsearch.client.node.NodeClient;
import org.elasticsearch.cluster.metadata.Metadata;
import org.elasticsearch.rest.BaseRestHandler;
import org.elasticsearch.rest.RestRequest;
import org.elasticsearch.rest.action.RestToXContentListener;
import org.elasticsearch.xpack.core.ml.action.DeleteForecastAction;
import org.elasticsearch.xpack.core.ml.job.config.Job;
import org.elasticsearch.xpack.core.ml.job.results.Forecast;
import org.elasticsearch.xpack.ml.MachineLearning;

import java.io.IOException;
import java.util.List;

import static org.elasticsearch.rest.RestRequest.Method.DELETE;

public class RestDeleteForecastAction extends BaseRestHandler {

    @Override
    public List<Route> routes() {
        return List.of(
            new Route(DELETE, MachineLearning.BASE_PATH + "anomaly_detectors/{" + Job.ID.getPreferredName() + "}/_forecast/"),
            new Route(DELETE, MachineLearning.BASE_PATH + "anomaly_detectors/{" + Job.ID.getPreferredName() +
                "}/_forecast/{" + Forecast.FORECAST_ID.getPreferredName() + "}")
        );
    }

    @Override
    public String getName() {
        return "ml_delete_forecast_action";
    }

    @Override
    protected RestChannelConsumer prepareRequest(RestRequest restRequest, NodeClient client) throws IOException {
        String jobId = restRequest.param(Job.ID.getPreferredName());
        String forecastId = restRequest.param(Forecast.FORECAST_ID.getPreferredName(), Metadata.ALL);
        final DeleteForecastAction.Request request = new DeleteForecastAction.Request(jobId, forecastId);
        request.timeout(restRequest.paramAsTime("timeout", request.timeout()));
        request.setAllowNoForecasts(restRequest.paramAsBoolean("allow_no_forecasts", request.isAllowNoForecasts()));
        return channel -> client.execute(DeleteForecastAction.INSTANCE, request, new RestToXContentListener<>(channel));
    }
}

redpajama_set_name: RedPajamaGithub


text:
< 100 Objects That Shaped Public Health
Elixir Sulfanilamide
In January 1937, the world-changing use of penicillin to treat infections was still five years away. When an article in the Journal of the American Medical Association suggested the antimicrobial drug sulfanilamide might fight strep infections, several companies rushed to develop capsules and tablets and get them to market.
Among those entering the fray were Tennessee's S.E. Massengill Company, which whipped up a raspberry-flavored liquid "elixir" to meet demand. The solution included 10 percent sulfanilamide, 16 percent water, small amounts of raspberry extract, saccharin, amaranth and caramel — and 72 percent diethylene glycol, a chemical used as a moistening agent and in the production of resins and explosives.
The company shipped its miracle drug to hundreds of pharmacies all over the country. But soon people were dying — 105 by the end of the crisis. Diethylene glycol, it turned out, was a poison and the elixir was toxic.
At the time, the young U.S. Food and Drug Administration did not have the authority to check the safety of medications before they went to market. In the wake of the tragedy, the U.S. Congress passed the Federal Food, Drug, and Cosmetic Act of 1938, which established a new standard requiring pharmaceuticals to undergo an FDA safety review before they could be sold.
Explore Other Objects
Tamper-Resistant Cap
Big Gulp

redpajama_set_name: RedPajamaCommonCrawl


Interested in adding a new family member? Check our current available rescues. You can use the links for Pet Finder or Adopt a Pet to find those looking for their forever home.
All potential adopters are required to submit an application. Once approved, you will be contacted and a "meet and greet" will be scheduled for you and your family (all two-legged and four-legged individuals) to meet the rescue dog.
Save your completed adoption application and email it to the volunteer designated for the puppy of your choice or puppiesandmore@yahoo.com.


text:
Q: Create Objects from Json dynamically in python I need to know some if it is possible to create objects from JSON in this way:
Let's suppose I got 2 objects Human and Animal, Human will have as attributes name, age, sex and Animal name, legs, colour.
I store the information in a MongoDb and when I retrieve something using a query I receive the result in Json
human = {'name':'Bogdan','age':'21','sex':'male'}

animal = {'name':'Bob','legs':'4','colour':'brown'}

Ok. Now it comes the question .. I want to decode the Json using a function and create objects from them, something like this :
def object_decoder(object, jsonToConvert):

 return object(**json.loads(jsonToConvert))

It is a good practice to try an abstraction of this functionality, or it is better to make a object_decoder function for every Object Model.
Thank you!

A: sure
class Thing:
     def __init__(self,**kwargs):
         for k,v in kwargs.items():
             setattr(self,k,v)
     def __getitem__(self,item):
         return self.__dict__.get(item,None)
     def __repr__(self):
         return "<Thing: %s>"%self.__dict__

Thing(**json.loads('{"name":"Bogdan","age":"21","sex":"male"}'))

in reality you probably shouldnt really do this...unless you have a good reason to.
classes are typically used when you want to encapsulate some behaviors along with stateful data.
when you just have data then a dictionary is just fine(your example is only data so no real point in a class) ...

A: No need to decode, simply enable to construct the object from a dict, or better, if you can - use the keywords as the field names you're using in mongo.
Then, just use:
import ast
class Human(object):
    # same as mongo
    def __init__(self, name=None, age=None, sex=None):
        print name
        print age
        print sex
# you should be able to get these as dicts straight away...
human = {'name':'Bogdan','age':'21','sex':'male'}
human_obj = Human(**human)

I'm not sure what the object type you're mapping is, so I added what I thought. If you have a dict - it's best.


text:
All Places > Electronics Legislation > Global Conflict Minerals > Americas Conflict Minerals > Documents
Conflict Minerals - exploring the Dodd-Frank Act
Created by VictoriaJones on Jun 7, 2011 4:11 AM. Last modified by VictoriaJones on Jun 28, 2012 4:58 AM.
Background to Conflict Minerals in the Congo
Concern was raised some years ago in the USA regarding the ethics of using materials sourced from the Democratic Republic of the Congo (DRC) in the USA, particularly in the electronics manufacturing sector. The reason behind the concern was that the sale of this material by armed militias was funding the civil war in the region, leading to exploitation of and the use of violence against the local population. In April 2009, a Congo Conflict Minerals Act was introduced in the USA, to require electronics companies to verify and disclose their sources, but this legislation died in committee.
In the aftermath of the Enron and other financial scandals in the USA, a major amendment to the Securities Exchange act of 1934 was formulated. This new law, the Dodd–Frank Wall Street Reform and Consumer Protection Act of 2010, was passed by the US Senate on 20 May 2010 before being signed into law by President Barack Obama on 21 July 2010. Section 1502, generally referred to as the "Conflict Minerals Law", requires US companies to disclose the source of certain minerals in their product. The original 1934 act has been amended to include this text.
This document seeks to summarise the situation and the impact on industry.
DRC is in central southern Africa, and was formerly known as the Belgian Congo and later Zaire. There is a short seaboard on the Atlantic Ocean at the mouth of the Congo River, with the port of Matadi, there is no direct outlet to the Indian Ocean. The surrounding countries are Angola, Zambia, Tanzania, Rwanda, Burundi, Sudan, the Central African Republic (CAR) and Republic of Congo (a separate nation to the DRC).
Dodd-Frank Act requirements
The main intent of the Dodd-Frank Act is to reduce the use of so-called "conflict minerals" from DRC and associated sources through a regime of public reporting and scrutiny which pressurise producers to take proper responsibility for their supply chains. The main outcome of this initiative will be an increased knowledge of the supply chain, and knowledge of where materials originate. Similar approaches have been successful in other industry sectors particularly forestry, diamonds (The Kimberley Process) and clothing.
The Securities and Exchange Commission (SEC) has defined "conflict minerals" as:
(A) columbite-tantalite (coltan), cassiterite, gold, wolframite, or their derivatives; or (B) any other mineral or its derivatives determined by the Secretary of State to be financing conflict in the Democratic Republic of the Congo or an adjoining country.
'Conflict minerals' and their use
The table below shows the source and some of the more commonplace uses for these materials, together with an approximation of the proportion of the mineral arising from the DRC.
Ore / Element
Examples of uses
Sources and comments
Approx % world supply from DRC (2007)
Cassiterite / Tin Solders, solderable coatings, corrosion resistant coatings, bronze alloys, glass additive, flame retardants, biocides
Many other sources outside this geography.
Major Source (2007) :
P R China : 43%
Wolframite / Tungsten
High temperature applications (lamp and electron microscope filaments, engines, lubricants, welding, photomultiplier tubes), hard alloys (e.g. turbine blades), tungsten carbide drill bits etc., munitions, radiation shielding, metal coatings on ceramics and on silicon ICs.
P R China 85.9%
Rwanda*
Colmubite-Tantalite (Coltan) / Tantalum
Tantalum capacitors, high refractive index glass, power resistors, high temperature alloys, corrosion resistance.
Few alternative sources outside this geography.
Australia 60.7%
9% DRC*
9% Rwanda*
Niobium Niobium alloys including steels, superconducting alloy with titanium or tin is used in MRI and other instruments with superconducting magnets. Pacemakers, sodium lamps and arc welding.
Brazil 53.4%
Various / Gold
IC wire bonding, plating, electric contacts and connectors.
South Africa 10.8%
Colbaltite etc / Cobalt
Lithium NiCd and NiNH batteries, hard and corrosion resistant plating, moisture indicators, special alloys (e.g. turbine blades), Alnico magnets.
Not explicitly named but is likely to qualify as an "other mineral".
~ 35%
~ 41% *
Source : USGS unless marked with * in which case EU Communication COM(2011) 25 final, 2 Feb 2011 for 2008/9.
Many of these types of electronic components come from China and one source of these raw materials will inevitably be the DRC. Nickel and copper are also mined in the DRC so could be added to the list.
Detailed compliance standards and expectations of the SEC are expected to be published during Q4 2011, with the first reporting period being for the fiscal year ending Q4 2012.
The essential requirement of the Dodd-Frank Act, Clause 1502 is to require any organisation placing products on the market in the US to declare whether "conflict minerals", originating from the DRC and adjoining countries are present. Thus companies that use the defined materials need to be able to show their origins; and as the national borders are somewhat leaky, the Act requires import from adjacent countries as well as the DRC to be recorded. For example, it is thought that Rwanda exports about 5 times the amount of tin than is actually mined in that country, which may not be immediately apparent from information received up the supply chain.
A company trading in the USA will be affected by this Act if :
(a) It files reports with the SEC under the Securities and Exchange Act of 1934
(b) Its use of the minerals is "necessary to the functionality or production" of the products.
The requirements are applicable to any organisation (or person), called an "issuer", using "conflict minerals" in their products based in the US whatever the size. This could include parties who:
Are the brand owner of a product.
Contract to have the product manufactured specifically for themselves.
Have influence over the "product's manufacturing".
This would therefore include parties who have design authority over a product or buy in large volume which is often the case for large retailers and distributors for example.
Thus in order to comply a company will need a detailed and extensive knowledge of its supply chain. It would be prudent for any company having a legal entity in the US to conduct and document a "due diligence" exercise of their supply chain with regard to the origins of any of the materials specified in Section 1502.
The SEC will also be looking for evidence of an independent 3rd Party audit of the process and results.
This declaration would be on an annual basis both to the SEC and on the company's public web site. Where "conflict minerals" might be used they must also report to the SEC the steps they are taking to identify the source and eradicate them.
Inclusion in the annual report to the SEC applies where "conflict materials" may be present and is based on a "reasonable country of origin inquiry". It is not stated what is reasonable but clearly the process used to define this for all products should be documented by any party to whom this Act may apply.
Where "Conflict Minerals" did not originate in DRC countries
Where it cannot be shown that "Conflict Minerals" did not originate in DRC countries
Disclose the determination that "Conflict Minerals" did not originate in DRC countries. Disclose this conclusion and Note that the "Conflict Minerals" Report is furnished as an exhibit (e.g. annex) to the annual report.
Disclose the "reasonable country of origin inquiry" process used in reaching this determination. Provide a "Conflict Minerals" Report.
Make the disclosure regarding this determination available on its Internet website. Make available the "Conflict Minerals" Report on its Internet website and disclose this fact.
Provide the Internet address of that site. Provide the Internet address of that site.
Maintain records demonstrating that its "conflict minerals" did not originate in the DRC countries
Documentation beyond the "Conflict Minerals"
Report is not stated as required but clearly would be prudent.
The "conflict minerals" report would be required to provide evidence of the due diligence measures taken and detailed information on the supply chain such as the facilities used to process the "conflict minerals" and efforts taken to locate the source. Showing the material to be sourced from scrap or recycled materials is one way of showing its origin. The report would be subject to audit by an independent third party according to a defined standard, which has yet to be elaborated.
While there are currently no legal requirements covering the use of "conflict minerals" within the EU, this has been a matter of scrutiny within the Commission and Parliament for many years. Recently the UK Chancellor of the Exchequer and Business Secretary raised this at the G20 Finance Minister's Meeting early in 2011, and are pushing for European legislation mirroring the requirements of the Dodd-Frank Act in the USA. As yet we have not seen any legislative proposals tabled however.
Implications of the Dodd-Frank Act and next steps for industry
While the Dodd-Frank fact applies only to publicly traded companies in the US, it will of necessity impact their supply chains as they seek better information on "conflict minerals" from their suppliers. It is also very likely to add to pressure on companies formally outside of scope to up their game both from customers and lobby groups. Companies will be asked serious questions by their customers and other parties, and as such they should develop and implement a plan to address "conflict minerals" as a matter of some urgency.
The requirements for meeting "conflict mineral" obligations are very similar to those of meeting the EU RoHS Directive or the REACH Regulation, in that in the event of enforcement action, an audit trail is necessary to be able to demonstrate to the authorities that a process of due diligence has been followed. A robust approach to showing due diligence often makes reference to and uses best practice.
OECD Guidance (Organisation for Economic Co-operation and Development)
The OECD has published guidance on responsible supply chain management6. The guidance was developed as a collaborative project involving the countries where "conflict minerals" arise, industry and the UN, therefore it carries significant credibility. The guidance advocates a five step approach to due diligence:
1. Establish strong company management systems.
2. Identify and assess risk in the supply chain.
3. Design and implement a strategy to respond to identified risks.
4. Carry out independent third-party audit of supply chain due diligence at identified points in the supply chain.
5. Report on supply chain due diligence.
Options for industry in addressing conflict minerals
Industry has a number of options available in addressing the "conflict minerals" problem:
This approach is essentially untenable for a major supplier of products which, in general worldwide, may make use of "conflict minerals". Companies will come under increasing pressure to respond from its customers and investors.
Industry to request information on "conflict mineral" compliance from its suppliers: This is a very difficult process and the response and quality of response is frequently low even for relatively straightforward requirements like RoHS. The response for "conflict minerals" is likely to be even poorer. Companies should also incorporate "conflict minerals" in supply chain audits.
Gathering essential information about conflict mineral status
As gaining information from the upstream supply chain is likely to be long and somewhat convoluted, this activity should be started as soon as is practicable.
Companies should initiate a query of suppliers about the presence or otherwise of the "conflict minerals" in their product, and their country of origin; perhaps adapting the EICC supply chain tool for this purpose.
1. Modify your product database to allow the recording of key data:
a. The request to the supplier; date sent, person sent to, etc
b. A number of follow up contacts made, with dates and responses
c. The response from the supplier (date received, how it was validated, follow-up actions)
d. Which "conflict minerals" may be present in the product, and in what proportion
e. The country of origin of the "conflict minerals" and any additional pertinent details on the source.
f. Steps being taken by the supplier to reduce the risk of "conflict minerals" being present.
2. As is the case with RoHS compliance, any supplier documentation should be signed off by a senior member of their management team.
3. The OECD guidance recommends retaining documentary evidence for a minimum period of 5 years.
Find out more Conflict Mineral information.
Register with element14 to be able to receive the latest Conflict Mineral legislation updates
Found the information, now get the part! Visit our online store
Got an opinion you want to share? Leave your comments below
Dodd Frank Act Conflict Minerals legislation September 2011 v2.pdf 1.3 MB
Categories: Tags: none (add) legislation, conflict_materials, congo, dodd_frank, conflict_minerals, legislation_ebook, legislation_all, dodd_frank_act, dodd-frank, conflict_minerals_act, drc_conflict, conflict_metals, sec_conflict_minerals, conflict_minerals_legislation, africa_dfc, conflict_drc, conflict_in_the_drc, conflict_minerals_law, conflict_minerals_dodd_frank, conflict_minerals_sec, conflict_minerals_trade_act, drc_conflict_minerals, dodd_frank_act_conflict_minerals, conflict_minerals_dodd-frank, what_is_dodd_frank, dodd_frank_bill, dodd_frank_legislation, cm_guidance, conflict_mineral_ebook, americas_guidance
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Conflict Minerals– Why no SEC rule yet?
Congo Conflict Minerals: California passes state bill


I am curious if I should sell my Eventide Timefactor and possibly replace with GFI Specular Tempus?
How about getting a H9 to replace the Time Factor?
I have to admit the GFI Specular Tempus does sound vey cool though in the online demos, and I like have delay and reverb in one unit!


I'd never imagined Mom leaving and not coming back.
After that, the measuring started. Just occasionally. Grandma said it meant he was thinking of Mom. Then Grandpa read somewhere how Giants have a lot of twins, and the measurements turned weekly. We got a long letter with a picture of Mom and her new/old boyfriend all curled up to keep his head in the frame. Grandpa deleted it from Grandma's inbox, and the measurements turned daily.
Around two a.m., she finds us in the hallway. Maybe our flashlight beams seeped under their door, maybe she smelled the wet paint—Grandpa's dead to the world when he's asleep, but Grandma's always shown up for puke or tears or growing pains since Mom left.
He and Grandma are yelling in the yard, a paint bucket tipped over between them. She tries to push past him toward the house, and Grandpa shouts, spits, red-faced. He's no giant and he's frail, but he's still much bigger than Grandma.
Originally appeared in Fireside Fiction in September 2017.
Allison Mulder grew up in the Midwest, and enjoys cheesy jokes and eldritch horrors in equal measure. Some of her interests include anime, videogames, podcasts, and the creepiest creatures that the cold, black depths of the ocean have to offer. Her stories have appeared in Fireside Fiction, Escape Pod, InterGalactic Medicine Show, and more. You can find them at allisonmulder.wordpress.com, though Allison herself is more easily found on Twitter as @AMulderWrites.


text:
Q: Running SikuliX Script on VPS while computer off? I'm trying to run a SikuliX Script 24/7 on a VPS but I do not want to leave my computer on all the time while it runs. I know sikuli requires that you have a monitor, so I am not sure how I can have it run while my VPS isn't even open on my computer.

A: If your VPS is running Linux you could use a tool such as Xpra, it allows for an X application to be run even when the client disconnects. It can also be used with a nested X server like Xephyr to run a full remote desktop that persists disconnects.
On Windows Server this this superuser page details how to allow a Windows Server RDP session to persist even after the client has disconnected.


text:
Q: A word for alertness for the future course of events Being alert, though not necessarily, usually indicates, perhaps subconsciously taken as, alertness to present. Say I am playing a game and being alert to what might go wrong in the coming moves. Is there a word specifically stressing such state of mind?

A: I don't think words like clairvoyant or prescient quite do it - they imply more certainty in my mind. Foresightful might capture the sense better though.

A: It sounds like anticipate

: to think of (something that will or might happen in the future)

Merriam-Webster
The adjectival form is anticipatory

A: Premonition or Precognition would fit.

*

*Premonition is a feeling of anticipation of or anxiety over a future event; presentiment - Example : He had a vague premonition of danger.

*Precognition is the knowledge of a future event or situation, especially when this comes from a direct message to the mind, such as in a dream or by extrasensory means, rather than by reason
Also Foreknowledge (awareness of something before it happens or exists).

A: Say I am playing a game and being alert to what might go wrong in the coming moves.

You are on the ball.

Alert and efficient or effective:
      If you don't get on the ball, you'll be fired.
Indicating intelligence or ability:
      The tests show your students don't have much on the ball.
The new manager has a lot on the ball.
(Random House)


SANTA FE � Governor Bill Richardson and First Lady Barbara Richardson, along with the New Mexico Arts Commission, have announced seven individuals and a high-profile classical radio station as recipients of the 2008 Governor�s Awards for Excellence in the Arts.
Harry Benjamin of Silver City for Painting; Tammy Garcia of Santa Clara Pueblo for Visual Arts � Pottery, Sculpture and Glass; Jack Loeffler of Santa Fe for Ethnomusicology and Writing; Noel Marquez of Artesia for Painting and Mural Artistry; Eugene Newmann of Ribera for Painting; Alice �Ali� MacGraw of Tesuque � Major Contributor to the Arts; Eileen A. Wells of Santa Fe � Major Contributor to the Arts; and KHFM Radio 95.5 of Albuquerque � Major Contributor to the Arts.
The 2008 Governor�s Arts Awards ceremonies will be held on Friday evening, September 12, 5:15 to 7:00 pm at the St. Francis Auditorium, Museum of Fine Arts in Santa Fe. The ceremony is preceded by an afternoon reception and exhibition opening, 3:00 � 4:30 pm, in the Governor�s Gallery, 4th Floor, State Capitol. Both the awards ceremony and gallery reception are free and open to the public.
The Governor's Arts Awards were established in 1974 by Governor Bruce King and First Lady Alice King to celebrate the enormous roles � both economic and cultural � that artists, craftspeople and arts supporters play in the life of New Mexico. During its 34-year existence, a diverse and prestigious list of painters, weavers, sculptors, dancers, musicians, storytellers, poets, actors, playwrights and potters have been honored. Past awardees include Georgia O�Keeffe, Laura Gilpin, Max Evans, Wilson Hurley, Joy Harjo, Bill Mauldin, John Nichols, Pablita Velarde and Cipriano Vigil.
Nominations for the awards are invited each year from arts groups and interested New Mexicans. All nominations are reviewed by a committee of the New Mexico Arts Commission, which sends its recommendations to the full Commission and to the Governor. The 2008 Awards Selection Committee consisted of Charmay Allred of Santa Fe as chairperson, Janice Spence of Hobbs, John Rohovec of Silver City, Glenn Cutter of Mesilla, Karen Cotter of Albuquerque and Chuck Zimmer, interim manager of the state public art program for New Mexico Arts. Loie Fecteau, executive director of New Mexico Arts, and media manager Virginia Castellano served on the committee in a nonvoting capacity.


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Oslo Freedom Forum (OFF) is a series of global conferences run by the New York-based non-profit Human Rights Foundation under the slogan "Challenging Power". OFF was founded in 2009 as a one-time event and has taken place annually ever since. One of the key objectives of the conferences is to bring together notable people, including former heads of state, winners of the Nobel Peace Prize, prisoners of conscience, as well as of other public figures in order to network and exchange ideas about human rights and exposing dictatorships.

The main OFF conference is held annually in central Oslo, Norway, while satellite events have been held in San Francisco and New York, and on college campuses in the United States. OFF talks are live-streamed and consist of lectures and panel discussions taking place in front of a live audience.

OFF was founded by human rights activist Thor Halvorssen.

Events
2009
The inaugural Oslo Freedom Forum, titled "The Nobility of the Human Spirit and the Power of Freedom", featured more than 30 speakers with an emphasis on the importance of literature in advancing the cause of freedom. Participants included Nobel Laureate Elie Wiesel; Czech playwright and politician Václav Havel; Kurdish rights advocate Leyla Zana; and Tibetan former political prisoner Palden Gyatso.

Norwegian daily Klassekampen called the conference an "impressive assembly of people." The Wall Street Journal described OFF as "unlike any other human-rights conference... the emphasis was on promoting basic rights in all nations at all times."

2010
The 2010 Oslo Freedom Forum, organized around the theme "From Tragedy to Triumph", featured participants from more than 40 countries and six continents. The event focused on the progress made in the realms of civil liberties and freedoms over the past century, while highlighting the innovation of modern-day advocates—activists, policy makers, world leaders, and media entrepreneurs.

The Economist described the 2010 forum as "A spectacular human-rights festival… on its way to becoming a human right equivalent of the Davos economic forum."

2011
The 2011 Oslo Freedom Forum took place under the theme "Spark of Change", and featured speakers from across the world who gave presentations on an array of topics, from an analysis of some of the world's most prominent dictatorships, to a look at the impact that a single individual can have on the world.

The Economist described the 2011 OFF as "a glittering gathering of veterans of human-rights struggles." Peter Thiel (of Facebook, PayPal, and LinkedIn) praised the Oslo Freedom Forum "because [its] focus on dissidents engages the intellectual debate as well as the moral cause."

2012
Titled "Out of Darkness, Into Light", the fourth annual Oslo Freedom Forum explored numerous topics, including a spotlight on the many forms of modern-day slavery; exposés on how Western public relations agencies, IT firms, and arms companies support dictatorships; the drug war's impact on human rights; a focus on the burgeoning democracy movement in Russia; perspectives on fighting poverty through individual rights; an examination of global censorship; and a discussion on the state of the Arab uprisings. Supporters included the City of Oslo, the Norwegian Ministry of Foreign Affairs, Amnesty International, the Thiel Foundation, Fritt Ord, the Nobel Peace Center, and the Norwegian Helsinki Committee.

The Daily Beast described the forum as a conference of "do-gooders conspiring to stir up trouble," while The Guardian called it "a Davos for Revolutionaries."

2013
The 2013 Oslo Freedom Forum was themed "Challenging Power" and centered on a range of topics, such as the art of dissent, asymmetric activism, new tools for rights advocates, the power of media, women under Islamic law, and the threat of authoritarian regimes with façade capitalism.

2013 speakers included Chinese dissident Chen Guangcheng; recently escaped blogger Bahraini Ali Abdulemam; Peruvian Nobel laureate Mario Vargas Llosa; Zimbabwean artist Owen Maseko; Tibetan prime minister Lobsang Sangay; creator of the Magnitsky Act Bill Browder; Palestinian journalist Asmaa al-Ghoul; Malaysian lawyer and democracy advocate Ambiga Sreenevasan; and Serbian nonviolent resistance leader Srđa Popović.

The conference culminated with the presentation of the Václav Havel International Prize for Creative Dissent. The 2013 laureates were Syrian cartoonist Ali Ferzat, North Korean democracy activist Park Sang Hak, and Cuban civil society group Ladies in White—represented by their leader Berta Soler.

The conference was chronicled in VICE, BuzzFeed, El País, El Mundo, Aftenposten, and Verdens Gang. Speakers were profiled in The Atlantic, CNN, The Economist, The New York Times, CNET, The Wall Street Journal, NPR, Dagens Næringsliv, and Finansavisen. BuzzFeed described the conference as "an internationalist networking party where dissidents trade tips on overthrowing authoritarian regimes," while Al Jazeera characterized it as "an annual conference that gives the people who challenge repressive regimes a platform to speak."

The forum ended with the presentation of the second Václav Havel Prize for Creative Dissent to Ali Ferzat, a Syrian political cartoonist, the Ladies in White (from Cuba) and Park Sang Hak, a North Korean democracy advocate.

2014
The 2014 Oslo Freedom Forum included Egyptian satirist Bassem Youssef; Harvard cognitive scientist Steven Pinker; Cuban blogger Yoani Sánchez; American actor Jeffrey Wright; Ukrainian pro-democracy activist Yulia Marushevska; iconic Turkish protestor Erdem Gündüz, as well as Marcela Turati Muñoz, Yeonmi Park, Hyeonseo Lee, Iyad El-Baghdadi, Ti-Anna Wang, Suleiman Bakhit, Jamila Raqib, Nadezhda Tolokonnikova, Maria Aljokhina and Mikhail Khodorkovskij.

The New York Times called the Oslo Freedom Forum the place where "the world's dissidents have their say."

2015
The 2015 Oslo Freedom Forum took place on 25–27 May and was described by The Financial Times as a "Davos for dissidents." Speakers included Charlie Hebdo columnist Zineb El Rhazoui, North Korean Ji Seong-ho, Afghan entrepreneur Saad Mohseni, and Twitter vice president Colin Crowell.

2016
The 2016 Oslo Freedom Forum took place on 23–25 May 2016. Speakers included Afghan entrepreneur Roya Mahboob and Russian opposition activist Vladimir Kara-Murza.

2017
The 2017 Oslo Freedom Forum took place on 23–24 May 2017. Speakers included Norway's Prime Minister Erna Solberg, the Maldives' first democratically elected president, Mohamed Nasheed, Peter Thiel, and American Nobel Laureate and director of the International Campaign to Ban Landmines, Jody Williams. A second event took place on 19 September 2017, in New York City, parallel to the 72nd session of the United Nations General Assembly. The event took place at Alice Tully Hall, Lincoln Center, and was considered a "one-day" version of the event typically held in Oslo. Speakers included among others former World Chess Champion and chairman of the Human Rights Foundation Garry Kasparov, Iranian dissident Marina Nemat, Russian democracy activist Vladimir Kara-Murza, Serbian political activist Srđa Popović and Venezuelan democracy activist Wuilly Arteaga.

2018
The Oslo Freedom Forum took place from 28 to 30 May 2018. Speakers included Clare Rewcastle Brown, Lebo Mashile, Asma Khalifa, Galia Benartzi, Mu Sochua, Leyla Yunus, Tiff Stevenson, Antonio Ledezma, Netiwit Chotiphatphaisal, Fang Zheng, Wael Ghonim, Jason Silva, Rick Doblin, Maziar Bahari and Emmanuel Jal.

2019
In 2019, The Oslo Freedom Forum held a one-day event in Taipei. Speakers included Denise Ho Thae Young-ho and Thai education reformer and activist Netiwit Chotiphatphaisal.

2020
Due to the coronavirus pandemic in 2020, the forum was held online. Speakers included Park Eun-hee,

Participants

San Francisco Freedom Forum
Iconic Burmese pro-democracy leader Aung San Suu Kyi headlined the 2012 San Francisco event, which featured talks exploring various paths to freedom. Other speakers included: Saudi women's rights pioneer Manal al-Sharif; conflict psychologist Justine Hardy; Iranian author and former prisoner of conscience Marina Nemat; Slate editor and author William J. Dobson; drug policy reformer Ethan Nadelmann; Chinese scholar and former political prisoner Yang Jianli; Ghanaian economist George Ayittey; Moroccan journalist Ahmed Benchemsi; and Kazakhstani theater director Bolat Atabayev.

As part of her first trip to the United States after more than 20 years under house arrest in Burma, Suu Kyi accepted the 2012 Václav Havel International Prize for Creative Dissent.

The 2016 San Francisco Freedom Forum took place at the Regency Center in the last week of September and included Rosa Maria Paya; Zineb El Rhazoui, Danilo Maldonado Machado, Lee Hyeon-seo, Roya Mahboob, Yulia Marushevska, Abdalaziz Alhamza, and Kimberley Motley.

College Freedom Forum
The first College Freedom Forum took place at Tufts University on 5 November 2013. Speakers included Iranian author and former prisoner of conscience Marina Nemat; Moroccan journalist and media entrepreneur Ahmed Benchemsi; Equatoguinean human rights lawyer Tutu Alicante; president and CEO of Liberty in North Korea (LiNK) Hannah Song; and Egyptian journalist Abeer Allam.

The second College Freedom Forum took place at the University of Colorado Boulder on 11 March 2014. Speakers included Chinese civil rights activist Chen Guangcheng; Ugandan LGBT rights advocate Kasha Jacqueline; Bahraini human rights activist Maryam al-Khawaja; and North Korean defector and democracy advocate Park Sang Hak. Talks were followed by a moderated question and answer session with the audience. The event was a joint initiative of the Human Rights Foundation (HRF) and the University of Colorado Boulder Distinguished Speakers Board and Cultural Events Board.

The third College Freedom Forum took place at Yale University on 26 March 2015. Speakers included Iranian author and former prisoner of conscience Marina Nemat; North Korean defector Yeonmi Park; American journalist William Dobson; and Serbian expert on nonviolent resistance Srđa Popović. Talks were followed by a question and answer session with the audience. The event was a joint initiative of the Human Rights Foundation (HRF) and Yale ThiNK (There is Hope in North Korea).

References

External links

Human rights organisations based in Norway
International conferences in Norway
Annual events in Norway
Recurring events established in 2009
2009 establishments in Norway

redpajama_set_name: RedPajamaWikipedia


text:
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text:
Justia Patents US Patent Application for RADIOLABELED BETA-GALACTOSIDASE SUBSTRATE FOR PET IMAGING OF SENESCENCE Patent Application (Application #20190381198)
RADIOLABELED BETA-GALACTOSIDASE SUBSTRATE FOR PET IMAGING OF SENESCENCE
Aug 21, 2019 - Eberhard Karls Universitaet Tuebingen Medizinische Fakultaet
The present invention relates to novel compounds useful for visualizing cell senescence in vitro and in vivo, the preparation of said compounds and their use. In particular, the present invention pertains to novel hexose and particularly galactose derivatives which are useful as senescence tracers in vitro and in vivo.
Latest Eberhard Karls Universitaet Tuebingen Medizinische Fakultaet Patents:
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CROSS REFERENCES TO RELATED APPLICATIONS
This application is a continuation of international patent application PCT/EP2018/054356, filed on Feb. 22, 2018 designating the U.S., which international patent application has been published in English language and claims priority from German patent application DE 10 2017 103 600.8, filed on Feb. 22, 2017.
The present invention relates to compounds useful for visualizing cell senescence in vitro and in vivo, the preparation of said compounds and their use. In particular, the present invention pertains to novel hexose and particularly galactose derivatives which are useful as senescence tracers in vitro and in vivo.
Cell senescence is the biological process by which cells exit the growth cycle. Cell senescence was first characterized in fibroblasts, which could only be subjected to a limited number of passages, before growth became permanently arrested. This phenomenon is named the Hayflick limit and serves to explain the physiological course of aging. It is accompanied by distinct change in metabolic pathways (Roninson, E. B., Tumor Cell Senescence in Cancer Treatment, Cancer Res., 2003, 63:2705-2715).
Senescent cells exhibit a senescence associated secretory phenotype, containing pro-inflammatory cytokines and growth factors. In certain instances, the removal of senescent tissues can convey great benefits to a patient. Senescence is recognized to play an important role in cancer treatment and therapy resistance. Treatment-associated senescence marks as stable clinical end point, and thus can be a measurement of chemotherapeutic success. The detection of senescence cells might also offer diagnostic opportunities for detecting pre-neoplastic lesions (Roninson, E. B., Tumor Cell Senescence in Cancer Treatment, Cancer Res., 2003, 63:2705-2715;and Campisi, J. and d'Adda di Fagagna, F., Cellular Senescence: when bad things happen to good cells, Nature Reviews Molecular Cell Biology, 2007, 8:729-740).
The most widely used surrogate marker for senescence cells is the senescence-associated beta-galactosidase (Roninson, E. B., Tumor Cell Senescence in Cancer Treatment, Cancer Res., 2003, 63:2705-2715). Such beta-galactosidase substrates have, however, the drawback that they merely may be used ex vivo and in vitro for following beta-galactosidase expression.
The present invention seeks to overcome the problems associated with prior art compounds.
There exists therefore an unmet need for further compounds that can act as tracers for cell senescence which may be used in vitro and in vivo at the same time. There is also a need in developing improved methods in the treatment of disorders related to cell senescence, such as cancer. A further objective of the present invention resides in alternative methods for selectively eliminating senescent cells in vivo. Still a further objective of the present invention is the provision of alternative means for determining the efficiency of cancer treatment.
The present invention is based on the finding that hexose derivatives, preferably galactoside derivatives and more preferably beta galactoside derivatives, carrying radioactive labels accumulate in senescent cells and can be used to reliably mark and detect said senescent cells both in vitro and in vivo. In vivo detection of senescent cells offers in turn improved access to such cells by surgery or therapy.
The present invention therefore provides a compound of the formula:
G-S-L,
wherein R is selected from H, substituted or unsubstituted C1 to C5 alkyl, substituted or unsubstituted C1 to C10 cycloalkyl, or substituted or unsubstituted C1 to C10 heterocycloalkyl,
and wherein * represents the binding site between G and S,
S is selected from the group consisting of
wherein X is independently selected from the group consisting of H, halogen, methyl halogen, OH and SH,
wherein Y is independently selected from the group consisting of C, S, N, and O, with the proviso that at least 3 C-atoms are present,
wherein # represents the binding site between S and L, and
L is
wherein R′ is CH2, NH, S, or O,
wherein n is 0 or 1,
wherein R″ is substituted or unsubstituted C1 to C5 alkyl,
wherein Z is a radioactive detectable label, a radioactive therapeutic residue, a chelator coordinating a radioactive detectable label or a chelator coordinating a radioactive therapeutic residue;
or a salt thereof.
It has been surprisingly found that the present compounds are highly and selectively enriched in senescent cells. Without wishing to be bound by theory, it is assumed that the rather small size of radioactive detectable labels in comparison to conventional non-radioactive labels, which is often based on the detection of bulky enzymes, such as horseradish peroxidase, alkaline phosphatase, or bulky dye compounds, offers improved access of the present compounds to the cells and the beta-galactosidase's binding site. Furthermore, the use of radioactive labels confers the advantage of highly enhanced sensitivity versus conventional labeling strategies. This is in part due to the high tissue penetration of the resulting gamma photons and very high sensitivity of the radio detectors used. This in turn allows for the denoted compounds to be translatable, in vivo, making them superior to nonradioactive alternatives. The present compounds thereby offer the possibility to quantitatively, selectively and sensitively detect senescent cells, particularly tumors, in vivo. This highly selective enrichment in senescent cells may be used in diagnosis or treatment of senescence related disorders by employing radioactive detectable labels or radioactive therapeutic residues in the present compounds.
Another advantage resides in the possibility to treat tumors with non-invasive techniques, i.e. by administering the present compounds exhibiting a radioactive therapeutic residue, which are almost exclusively incorporated in tumor tissue. Another advantage of the use of the present compounds resides in that they may be easily adapted to the changing requirements in the clinical field since the present compound may be easily provided with another radioactive detectable label or radioactive therapeutic residue.
The compounds of the invention may, depending on their structure, exist in stereoisomeric forms (enantiomers, diastereomers). The invention therefore also encompasses the enantiomers or diastereomers and respective mixtures thereof. The stereoisomerically uniform constituents can be isolated in a known manner from such mixtures of enantiomers and/or diastereomers.
It is clear that the salts preferred for the purposes of the present invention are pharmaceutically acceptable salts of the compounds of the invention. However, also encompassed are salts which are themselves not suitable for pharmaceutical applications but can be used for example for the isolation or purification of the compounds of the invention.
Salts in general are composed of related numbers of cations and anions so that the product is electrically neutral. It will be appreciated that a respective counterion is subject to the preparation process of the salt and needs not to be expressively mentioned.
Pharmaceutically acceptable salts as well as the preparation thereof are well-known in the art. Types and preparation of such pharmaceutically acceptable salts may be derived from Stahl, P. H. and Wermuth, C. G., Handbook of Pharmaceutical Salts: Properties, Selection and Use, Weinheim/Zürich: Wiley-VCH/VHCA, 2002.
Examples of pharmaceutically acceptable salts of the present compounds include salts of inorganic basis like ammonium salts, alkali metal salts, alkaline earth metal salts, salts of organic basis, or salts with basic amino acids. Also included are inorganic acids or salts with acidic amino acids. Preferred pharmaceutically acceptable salts encompass
It will be appreciated that the compounds of the present invention as well as the salts thereof may be present in form of solvates. In such a case, the present compounds or salts, particularly the pharmaceutically acceptable salts, thereof form in the solid or liquid state a complex by coordination with solvent molecules. Hydrates are a specific form of solvates in which the coordination displays with water.
If the compounds of the present invention may occur in tautomeric forms, the present invention encompasses all tautomeric forms.
The term "C1-C5 alkyl" generally refers to branched or straight-chain alkyl, preferably (C1-C4)-alkyl, such as in particular methyl, ethyl, propyl, butyl, isopropyl, isobutyl and tert.-butyl.
The term "C1-C10 cycloalkyl" generally refers to saturated or partially unsaturated monocyclic or polycyclic ring comprising carbon and hydrogen atoms.
The term "C1-C10 heterocycloalkyl" means a non-aromatic monocyclic or polycyclic ring comprising carbon and hydrogen atoms and at least one heteroatom, preferably, 1 to 4 heteroatoms selected from nitrogen, oxygen, and sulfur. A heterocycloalkyl group can have one or more carbon-carbon double bonds or carbon-heteroatoms double bonds in the ring as long as the ring is not rendered aromatic by their presence. Examples of heterocycloalkyl groups include aziridinyl, pyrrolidinyl, pyrrolidino, piperidinyl, piperidino, piperazinyl, piperazino, morpholinyl, morpholino, thiomorpholinyl, thiomorpholino, tetrahydrofuranyl, tetrahydrothiofuranyl, tetrahydropyranyl, and pyranyl. Preferably, the heterocycloalkyl group is a monocyclic or bicyclic ring, more preferably, a monocyclic ring, wherein the ring comprises from 2 to 6 carbon atoms and from 1 to 3 heteroatoms. More preferably the molecular weight of a heterocycloalkyl residue has a molecular weight of ≤200 g/mol, such as ≤150 g/mol, 120 g/mol or ≤100 g/mol.
The term "substituted" as used herein refers to one or more residues connected to C1-C5 alkyl, C1-C10 cycloalcyl and C1-C10 heterocycloalkyl and/or a modification to the carbon chain by introducing one or more heteroatoms, such as N; S or O. A residue may be furthermore selected from the group consisting of halogen, methyl, ethyl or functional groups, such as a hydroxyl group, amino group or carboxyl group. The overall molecular weight of residues connected to particular C1-C5alkyl, C1-C10 cycloalcyl or C1-C10heterocycloalkyl is preferably 250 g/mol, more preferably 150 g/mol or 100 g/mol. Examples of preferred substituted to C1-C5 alkyl residues encompass compounds such as CH2—O—CH2, C2H4—O—CH2, CH2—O—CH2—O—CH2, CH2—O—CH2—O—CH2, C2H4—O—CH2—O—CH2, CH2—O—C2H4—O—CH2, C2H4—O—C2H4—O—CH2, C2H4—O—CH2—O—C2H4, CH2—O—CH2—O—CH2—O—CH2, C2H4—O—CH2—O—CH2—O—CH2, and CH2—O—C2H4—O—CH2—O—CH2.
The term "halogen" or halo refers to fluorine, chlorine, bromine or iodine; preferably fluorine, chlorine or bromine. In some embodiments the term halogen or halo preferably refers to 76Br, 75Br, 19F and 18F.
The term "methyl halogen" refers to a methyl group having a single fluorine, chlorine, bromine or iodine, or two or three halogens which are independently selected from fluorine, chlorine, bromine or iodine. In some embodiments the methyl halogen exhibits preferably 76Br, 75Br, 19F and 18F.
The term "radioactive detectable label" encompasses for instance 11C, 40K, 13N, 15O, 18F, 75Br, 76Br, 82Rb, 68Ga, 64Cu, 62Cu, 123I, 124I, 125I, 131I, 210At, 211At and 111In. Preferred radioactive detectable labels are 11C, 18F, 68Ga, 64Cu, and 124I. More preferred is 18F. The radioactive detectable label allows quantitatively and/or qualitatively assessment of senescent cells as well as their location particularly in vivo. Hence, senescent tissue may be properly identified and for instance surgically removed with high selectivity.
Alternatively, a complex containing such an atom in a coordinated form is encompassed. In such a case the complex may be also denominated a chelator coordinating a radioactive detectable label. A (coordination) complex consists of a central atom or ion, which is usually metallic, and a surrounding array of ligands or complexing agents. Usually the central atom or ion is the radioactive detectable label. Residue Z therefore encompasses the central atom or ion as well as the surrounding array of ligands or complexing agents. Examples of such chelators coordinating a radioactive detectable label are well known to the skilled person. Preferably such a chelator is an amino acid, preferably a proteinogenic/natural amino acid, such as histidine. Other suitable examples of chelators comprise DOTA, NOTA, NODAGA and desferrioxamine, such as desferrioxamine B.
The term "radioactive therapeutic residue" as used herein refers to an atom, such as 32P, 60Co, 89Sr, 186Re and 153Sm. Other examples of radioactive therapeutic residues encompass 125I and 131I, which have been already mentioned as examples for radioactive detectable labels, as well as 86Y, 177Lu and 67Cu. Alternatively, a complex containing such an atom in a coordinated form is encompassed. In such a case the complex may be also denominated a chelator coordinating a radioactive therapeutic residue. The use of the present compounds with therapeutic residues in treatment of disorders associated with cell senescence, particularly cancer, combines the advantage of target selectivity with that of being systemic, as with chemotherapy, and it may be used as part of a therapeutic strategy with curative intent or for disease control and palliation.
It will be appreciated that respective radioactive therapeutic residues may be generated and attached to the present compounds in the same manner as radioactive detectable labels. Preferably, therapeutic residue and detectable label are identical permitting diagnosis and treatment of disorders associated with cell senescence, particularly cancer.
In formulae, the group which is represented by G, the end point of the line adjacent to which there is a *, is not a carbon atom or a CH2 group but rather a component of the bond to the atom to which G is attached.
In its broadest form, group G having the formula
may be also denominated hexose derivative including allose, altrose, glucose, mannose gulose, idose, galactose or talose derivatives having residue R, respectively. The hexose derivatives are preferably D-isomers. The most preferred hexose derivative is a beta-D-galactopyranoside.
R is selected from H, substituted or unsubstituted C1 to C5 alkyl, substituted or unsubstituted C1 to C10 cycloalkyl, or substituted or unsubstituted C1 to C10 heterocycloalkyl, as indicated above. In case R is substituted or unsubstituted C1 to C5 alkyl, preferably unsubstituted Ci to C5 alkyl, more preferably methyl, prolonged retention time of the present compound in comparison to R═H may be expected due to an impeded enzymatic cleavage. Said compounds may be therefore present in higher levels within senescent cells.
In formulae, the group which is represented by S, the end point of the line adjacent to which there is a * or #, is not a carbon atom or a CH2 group but rather a component of the bond to the atom to which S is attached.
In formulae, the group which is represented by L, the end point of the line adjacent to which there is a #, is not a carbon atom or a CH2 group but rather a component of the bond to the atom to which L is attached. Residue L may be either Z, wherein Z is a radioactive detectable label, a radioactive therapeutic residue, a chelator coordinating a radioactive detectable label or a chelator coordinating a radioactive therapeutic residue. In said case, Z is directly attached to S. Alternatively, residue Z attached via a spacer to S. In this case the spacer has the formula
In the context of the present invention, the expression "have/contain" or "having/containing" designates an open enumeration and does not exclude other components apart from the expressly named components.
In the context of the present invention, the expression "consists of" or "consisting of" designates a closed enumeration and excludes any other components apart from the expressly named components.
In the context of the present invention, the expression "essentially consists of" or "essentially consisting of" designates a partially closed enumeration and designates preparations which apart from the named components only have such further components as do not materially alter the character of the preparation according to the invention.
When in the context of the present invention a preparation is described with the use of the expression "have" or "having", this expressly includes preparations which consist of said components or essentially consist of said components.
Radiology as used herein refers to any kind of apparatus or device, capable of producing a visual signal or an image upon detecting the present residue Z, i.e. a radioactive detectable label, a radioactive therapeutic residue, a chelator coordinating a radioactive detectable label or a chelator coordinating a radioactive therapeutic residue. Preferably the apparatus permits localization of residue Z within a mammal. Non limiting examples of radiology include positron-emissions-tomography (PET), which produces a three-dimensional image or map of functional processes in the body. The system detects pairs of gamma rays emitted indirectly by a positron-emitting radioisotope, which is introduced into the body on a metabolically active molecule, i.e. a substrate to beta-galactosidase. Images of metabolic activity in space are then reconstructed by computer analysis, which may be supported by a CT X-ray scan performed on the patient in the same session, preferably at the same time, and in the same device in order to obtain a three dimensional image enabling localization of tissue in which the prodrug is enriched. In positron emission a proton is converted via the weak force to a neutron, a positron and a neutrino. Isotopes, which undergo this so called beta plus decay, thereby emit positrons. Suitable positron-emitting radionuclides for this purpose include 11C, 40K, 13N, 15O, 18F, 75Br, 76Br, 89Zr, 82Rb, 68Ga, 62Cu and 64Cu, of which 11C and 18F are preferred. Other useful radionuclides include 123I, 124I, 125I, 131I, 210At, 211At and 111In. The present compounds may be also labeled with technetium and rhenium isotopes using known chelating complexes. Methods for the generation of radionuclides as well as radio labeling of compounds are well known to the skilled person. US 2007/0273308 and WO 2007/122488 pertain e.g. to the production of radionuclides. Radiolabeling is for example outlined in WO 2007/148089 and WO 2007/148083.
PET is preferably coupled with a computer tomography (CT). Such PET/CT devices enable quantitative detection and allocation of the signals detected to particular tissues, i.e. a localization of the radionuclides employed and hence of the prodrugs attached thereto. Function and operation of PET/CT as well as devices are well known to the skilled person. Other suitable techniques comprise single photon emission computed tomography (SPECT) and techniques based on nuclear magnetic resonance (NMR) wherein the quantum mechanical magnetic properties of an atom's nucleus is detected.
The radiology device employed is preferably a PET device, more preferably a PET/CT device or a PET/MR device.
In the Figures
FIG. 1 shows a motive explaining functioning of a radio-labelled hexose substrate;
FIGS. 2A-2C show the tracer 9 as a substrate of commercially available betagalactosidase;
FIGS. 3A-3B show stability of the tracer 9 under in-vivo conditions;
FIGS. 4A-4B show in vitro validation of tracer in HCT116 and Hras cells;
FIG. 5 shows uptake of tracer 9 in senescent versus control HCT116 tumors;
FIG. 6 shows tracer 9 uptake in senescent versus control Hras tumors; and
FIGS. 7A-7B show immunohistochemical staining from (a) HCT116 and (b) Hras driven tumor models.
EMBODIMENTS
According to a preferred embodiment of the present invention G is
R is as indicated above. Preferably R is H, thereby defining a beta-D-galactose residue, in particular a beta-D-galactopyranoside, which is the best possible sugar moiety for cleavage by beta-galactosidase.
According to another preferred embodiment of the present invention R is H or CH3, X is independently selected from H or methyl halide, Y is independently selected from C and N with the proviso that at least 5 C-atoms are present, R′ is NH, n is 1, and R″ is a linear ether having 2 to 5 C-atoms.
According to still another preferred embodiment of the present invention R is H, S is selected from the group consisting of
In said compounds, L may be as indiacted above.
In case L is R″ is preferably selected from the group consisting of CH2—O—CH2, C2H4—O—CH2, CH2—O—CH2—O—CH2, CH2—O—CH2—O—CH2, C2H4—O—CH2—O—CH2, CH2—O−C2H4—O—CH2, C2H4—O—C2H4—O—CH2, C2H4—O—CH2—O—C2H4, CH2—O—CH2—O—CH2—O—CH2, C2H4—O—CH2—O—CH2—O—CH2, and CH2—O—C2H4—O—CH2—O—CH2. R′ is CH2, NH or O, preferably NH. Even more preferably, Z is 18F. In case R′ is CH2, it is preferred that n is 0 and R″ is an unsubstituted Ci to C5 alkyl. More preferably, Z is 18F.
In case L is
Z is preferably 18F.
Particular preferred compounds of the present invention R is H, S is selected from the group consisting of
In said compounds, G is
wherein R is H.
More preferably, Z is 18F.
According to a preferred embodiment of the present invention the compound is
The second structure employing a spacer molecule is particularly suitably for comparatively large radioactive detectable labels, such as 123I, 124I, 125I, 131I, 210At, 211At and 111In, and large radioactive therapeutic residues, such as 89Sr, 186Re and 153Sm. It is however, preferred that Z is 18F.
According to another preferred embodiment of the present invention the radioactive detectable label is selected from the group consisting of 11C, 40K, 13N, 15O, 18F, 75Br, 76Br, 82Rb, 68Ga, 64Cu, 62Cu, 123I, 124I, 125I, 131I, 210At, 211At and 111In, preferably 11C, 18F, 68Ga, 64Cu, and 124I, more preferably 18F.
According to still another preferred embodiment of the present invention the radioactive therapeutic residue is selected from the group consisting of 32P, 60Co, 64Cu, 89Sr, 90Y, 177Lu, 186Re and 153Sm.
According to a preferred embodiment of the present invention the present compound is for use in surgery. The present compound is preferably employed in combination with at least one inert, non-toxic, pharmaceutically suitable excipient.
According to a preferred embodiment of the present invention the present compound is for use as a medicament. The present compound is preferably employed in combination with at least one inert, non-toxic, pharmaceutically suitable excipient.
According to another preferred embodiment of the present invention the present compound for use in a method for detecting cell senescence.
According to another preferred embodiment of the present invention the present compound is for use in a method of determining the efficiency of cancer treatment.
The cancer to be diagnosed and/or treated by the present compounds may be selected from prostate carcinoma, colorectal carcinoma, breast cancer, lung tumors, tumors of the male or female genitourinary system, malign melanoma, cervix and throat tumor/cervical tumor, malign lymphoma, neoplasia of the hematopoietic system and musculoskeletal tumors.
According to still another preferred embodiment of the present invention the method for detecting cell senescence comprises contacting cells with the present compound.
According to still another preferred embodiment of the present invention the method for determining the efficiency of cancer treatment comprises contacting cells with the present compound.
According to still another preferred embodiment of the present invention the method is performed in vivo.
According to still another preferred embodiment of the present invention the method is performed in vitro.
The above methods can be performed both in vivo, e.g. in a human patient for monitoring the efficiency of cancer treatment, or in vitro, e.g. for screening for new medicaments.
The present compound will preferably be administered parenterally. For this administration route, the present compounds may be administered in suitable administration forms. Parenteral administration can take place with avoiding of an absorption step (e.g. intravenous, intraarterial, intracardiac, intraspinal or intralumbar) or with inclusion of an absorption step (e.g. intramuscular, subcutaneous, intracutaneous, percutaneous or intraperitoneal). Administration forms suitable for parenteral administration are, inter alia, preparations for injection and infusion in the form of solutions, suspensions, emulsions, lyophilisates or sterile powders.
The present compounds may be converted into the stated administration forms. This can take place in a manner known per se by mixing with inert, non-toxic, pharmaceutically acceptable excipients. These excipients include inter alia carriers (for example microcrystalline cellulose, lactose, mannitol), solvents (e.g. liquid polyethylene glycols), emulsifiers and dispersants or wetting agents (for example sodium dodecyl sulfate, polyoxysorbitan oleate), binders (for example polyvinylpyrrolidone), synthetic and natural polymers (for example albumin), stabilizers (e.g. antioxidants such as, for example, ascorbic acid), colors (e.g. inorganic pigments such as, for example, iron oxides) and taste and/or odor corrigents.
The principle underlying the present invention is generally shown in FIG. 1. A radio-labelled glycosidase substrate, i.e. a compound of the present invention, is converted by a glycosidase, in particular beta-galactosidase, to the corresponding sugar and the radio-labelled alcohol. The present compound is shown with residue W indicative of the moieties forming a hexose derivative, such as a galactose derivative, preferably beta-D-galactose. The glycosidase, particularly the beta-galactosidase, is overexpressed and accumulated in senescent cells. The radiolabelled alcohol in turn is accumulated in acidic lysosomes and may be detected by a radiology device, such as a PET/CT device.
The compounds of the present invention may be generally prepared by the route indicated in reaction scheme I.
In the upper row of reaction scheme I, generation of compound 5 is shown. Compound 5 does not exhibit a radioactive label, i.e. F is 19F. Reaction I is conducted with HBF4, NaNO2 at a temperature of 0° C. The yield of compound 2 is 63% w/w. Reaction II is conducted with Ag2CO3 and at a temperature of 0° C. The yield of compound 4 is between 40 to 60% w/w. Reaction III is conducted with NaOMe and MeOH at a temperature in the range from 20 to 25° C. followed by reaction IV in presence of amberlite IR120.
In the lower row of reaction scheme I, generation of compound 9 (tracer 9), a compound according to the present invention, is shown. Reaction II is conducted with Ag2CO3 at a temperature of 0° C. Reaction VI is conducted with Ag2CO3 at a temperature of 0° C. Reaction V is conducted with 18F− and DMSO at a temperature of 150° C. for 5 min. Reaction VI is conducted with NaOMe/MeOH, or MeOH, Et3N, H2O (10:1:1)
The compounds of the present invention show a valuable range of pharmalogical effects which could not have been predicted. They are capable of marking senescent cells in vitro and in vivo. Particularly, in vivo marking is made to such an extent that senescent cells may be unambiguously identified in course of a surgical procedure which in turn allows targeted elimination of such cells, particularly cancer cells.
It is to be understood that the above description is intended to be illustrative only and not restrictive. Many embodiments will be apparent to those skilled in the art upon reviewing the above description. By way of example, the invention has been described preliminarily with reference to synthesis as well as diagnosis of tracer 9. It should be clear that all kinds of suitable detectable labels and therapeutic residues may be synthesized and attached to the present compounds. The scope of the invention should, therefore, be determined not with reference to the above description, but should instead be determined with reference to the appended claims, along with the full scope of equivalents to which such claims are entitled.
The percentage data in the following tests and examples are, unless indicated otherwise, percentages by weight; parts are parts by weight. Solvent ratios, dilution ratios and concentration data of liquid/liquid solutions are based in each case on volume. The statement "w/v" means "weight/volume". Thus, for example, "10% w/v" means: 100 ml of solution or suspension contain 10 g of substance.
Optimized radio synthesis utilizes a TRACERIab FX N Pro (GE). 18F is produced as hydrofluoric acid (HF) using a PETtrace cyclotron (GE). 18F labelling relied on the nucleophilic substitution of an aromatic nitro group. The synthetic route is described in reaction scheme I.
The following subcutaneous animal models are used for tracer evaluation.
A xenograft model of colorectal cancer cell line (HCT116) with therapy induced senescence, doxorubicin is used as a chemotherapeutic. Senescence is induced in HCT116 xenograft tumors by intravenous (i.v.) administration of doxorubicin (10 mg/kg). Through tail vain of the mice, 5 days later dynamic PET/MRT scans (1 h) with compound 9 are performed.
As the second model, transgenic liver progenitor cell line (Hras) was used for subcutaneous allografts in nude mice. The cell line expresses HrasG12v and p53shRNA under doxycycline (doxy) conditions. After the removal of doxycycline, the p53 mRNA can be restored and translated to p53 protein. The high amount of p53 triggers senescence. The animals bearing Hras tumors received doxy-water (0.2 mg/ml) for tumor development and 14 days, after removal of doxy-water, the PET/MRT scans are performed with compound 9.
Example 1 discloses synthesis of compounds according to reaction scheme I.
Synthesis of Compound 4
(2R,3S, 4S, 5R,6S)-2-(acetoxymethyl)-6-((2-fluoropyridin-3-yl)oxy)tetrahydro-2H-pyran-3,4,5-triyltriacetate, 4: A solution comprising 2,3,4,6-Tetra—O—acetyl-α-D-galactopyranosyl bromide 3 (1.0 g, 2.4 mmol), Ag2CO3 (0.74 g, 2.7 mmol), MS-4Å (5 g) and 2-fluoro-pyridin-3-ol 5 (0.33 g, 2.9 mmol) in anhydrous DCM (15 mL) is stirred overnight, in the dark, at room temperature, under an argon atmosphere. TLC analysis indicated the complete consumption of the starting bromide and the formation of a new nonpolar product. The solution is filtered through a celite, which was rinsed with DCM (3×10 mL). After removing the solvent under reduced pressure, the crude residue is purified directly using silica gel column chromatography, using an increasing gradient of EtOAc in PE. The product afforded is a colorless solid (0.51 g, 51%).
TLC: Rf=0.5 (EtOAc: PE=1:1); 1H NMR (600 MHz, Chloroform-d) δ 7.94 (d, J=4.6 Hz, 1H, ArH), 7.58 (t, J=7.8 Hz, 1H, ArH), 7.13 (dd, J=7.8, 4.8 Hz, 1H, ArH), 5.50 (dd, J=10.5, 7.9 Hz, 1H), 5.45 (d, J=3.4 Hz, 1H), 5.10 (dd, J=10.5, 3.4 Hz, 1H), 4.96 (d, J=7.9 Hz, 1H), 4.22 (dd, J=11.4, 6.8 Hz, 1H), 4.15 (dd, J=11.4, 6.3 Hz, 1H), 4.01 (td, J=6.8, 1.1 Hz, 1H), 2.19 (s, 3H, OAc), 2.11 (s, 3H, OAc), 2.04 (s, 3H, OAc), 2.02 (s, 3H, OAc); 13C NMR (151 MHz, CDCl3) δ 170.26 (COquart), 170.13 (COquart), 170.04 (COquart), 169.44 (COquart), 154.75 (d, J=239.9 Hz), 141.59 (d, J=13.4 Hz, ArC), 139.49 (d, J=25.5 Hz, ArC), 130.22 (d, J=3.5 Hz, ArC), 121.85 (d, J=4.3 Hz, ArC), 121.23 (CH), 101.12 (CH), 71.38 (CH), 70.5 (CH), 68.29 (CH), 66.70 (CH), 61.17 (CH2), 21.03 (OAc), 20.63 (OAc), 20.61 (OAc), 20.55 (OAc).
(2S,3R,4S,5R,6R)-2-((2-fluoropyridin-3-yl)oxy)-6-(hydroxyl-methyl)tetrahydro-2H-pyran-3,4,5-triol, 5: Catalytic sodium methoxide is added to a solution of 4 (0.5 g, 1.1 mmol) in dry methanol (5 mL). The solution is stirred at room temperature until TLC analysis revealed that the hydrolysis of the acetylated sugar is complete. Acetic acid (0.2 mL) is added, after which the solvents are removed under reduced pressure. The remaining solid was of high purity, but subsequent recrystallization from methanol afforded 6 as a highly pure crystalline solid (225 mg, 73%).
TLC: Rf=0.25 (EtOAc: MeOH=9: 1); 1H NMR (600 MHz, Deuterium Oxide) δ 7.79 (dd, J=8.1, 5.0 Hz, 1H, ArH), 7.72 (t, J=8.9, 8.1 Hz, 1H, ArH), 7.26 (dd, J=8.0, 5.0 Hz, 1H, ArH), 5.06 (d, J=7.8 Hz, 1H), 3.95 (dd, J=3.4, 1.8 Hz, 1H), 3.84-3.78 (m, 2H), 3.75-3.70 (m, 3H), 1.85 (dd, J=2.3, 1.1 Hz, 1H); 130 NMR (151 MHz, Deuterium Oxide) δ 153.70 (d, J=238.5 Hz, CF), 141.49, 139.72 (d, J=23.9 Hz, ArC), 139.29 (d, J=11.6 Hz, ArC), 128.11 (d, J=3.7 Hz, ArC), 122.75 (d, J=4.2 Hz, ArC), 101.24 (CH), 75.68 (CH), 72.41 (CH), 70.31 (CH), 68.36 (CH), 60.68 (CH2); HRMS (ESI): [M+Na]+(theor.)=298,06974, measured=298,06998
(2R,3S,4S,5R,6S)-2-(acetoxymethyl)-6-((2-nitropyridin-3-yl)oxy)tetrahydro-2H-pyran-3,4,5-triyltriacetate, 7: A solution comprising 2,3,4,6-Tetra—O—acetyl-α-D-galactopyranosyl bromide 3 (1.0 g, 2.4 mmol), Ag2CO3 (1.3 g, 4.8 mmol), MS-4Å (5 g) and 2-nitro-pyridin-3-ol 6 (1.4 g, 9.7 mmol) in anhydrous DCM (10 mL) is stirred over night, in the dark, at room temperature, under an argon atmosphere. TLC analysis indicates the complete consumption of the starting bromide and the formation of a new nonpolar product. The solution is filtered through a celite, which is rinsed with DCM (3×10 mL). After removing the solvent under reduced pressure, the crude residue is purified directly using silica gel column chromatography, using an increasing gradient of EtOAc in PE. The product 7 afforded is a colorless solid (0.7 g, 63%).
TLC: Rf=0.65 (EtOAc: PE=4: 1); 1H NMR (600 MHz, Chloroform-0 6 8.26 (dd, J=4.6, 1.4 Hz, 1H, ArH), 7.82 (dd, J=8.4, 1.4 Hz, 1H, ArH), 7.54 (dd, J=8.4, 4.6 Hz, 1H, ArH), 5.51 (dd, J=10.5, 7.9 Hz, 1H), 5.47 (dd, J=3.4, 1.2 Hz, 1H), 5.10 (dd, J=10.5, 3.4 Hz, 1H), 5.06 (d, J=7.9 Hz, 1H), 4.25 (dd, J=11.4, 6.1 Hz, 1H), 4.16 (dd, J=11.4, 6.1 Hz, 1H), 4.06 (ddd, J=6.1, 6.1, 1.3 Hz, 1H), 2.19 (s, 3H, OAc), 2.14 (s, 3H, OAc), 2.06 (s, 3H, OAc), 2.01 (s, 3H, OAc); 13C NMR (151 MHz, CDCl3) δ 170.21 (COquart), 170.04 (COquart), 170.01 (COquart), 169.32 (COquart), 150.42 (ArCquart), 144.25 (ArCquart), 142.91 (ArC), 129.86 (ArC), 128.29 (ArC), 100.94 (CH), 71.65 (CH), 70.31 (CH), 67.64 (CH), 66.58 (CH), 61.26 (CH2), 20.62 (OAC), 20.60 (OAC), 20.56 (OAC), 20.51 (OAC); HRMS (ESI): [M+Na]+(theor.)=493,10649 measured=493,10686.
Synthesis of Compounds 8 and 9
Radiolabeling of 7 proceeds readily in DMSO at 150° C. Azeotropically dried [K.2.2.2]+18F− fluoride complex is used, producing the acetylated 18F-labelled intermediate, which is purified using semi-preperative HPLC. The tracer was concentrated by trapping onto a 018 SPE cartridge, deprotected with NaOH (185 gt, 2.0 M) to produce tracer 9 and eluted with 10% ethanol in water (3 mL). It is appropriately formulated with HCl (195 gt, 2.0 M), NaHCO3 (500 μL, 1.0 M) and water (5 mL) to ensure subisotonic sodium concentration (0.1 M) and a final pH (7.5) that is biologically tolerated. The tracer is produced with a radiochemical purity of >98%, the decay corrected yield was 18.6+/−2.5% (n=10) with a molar radioactivity of 18.8±3.5 GBq* μmole−1 (n=5).
To assess whether tracer 9 is a substrate of beta-galactosidase, it is incubated at 30° C. for 30 minutes in citrate buffer of pH 5.5 containing commercially available betagalactosidase (Sigma). The reaction is stopped by adding 2 volume equivalents of acetonitrile, followed by 5 minutes of centrifugation. The supernatant is examined directly using radio-HPLC. The reaction produces the radioactive metabolite with the same retention time as the non-radioactive 2-fluoropyridinol. The expected radioactive metabolite is compared with the authentic non-radioactive standard (FIG. 2). In particular, compound 10 is radioactive 2-fluoropyridinol (FIG. 2a), compound 9 is tracer 9 (FIG. 2b), and compounds 1 and 2 are other compounds of reaction scheme I (FIG. 2c). Compound 11 is galactose.
The tracer 9 is incubated at 37° C. in mouse serum to assess its stability. After 1 hour, 2 volume equivalents of acetonitrile are added and the mixture is centrifuged for 5 minutes. The supernatant is examined directly using radio-HPLC. Serum stability of tracer 9 after 1 h (FIG. 3a) vs tracer 9 (FIG. 3b) is derivable from FIG. 3. The experiment reveals a few minor radioactive metabolites, but tracer's integrity remains largely intact.
In-vitro evaluation of compound 9 is shown in FIG. 4. HCT116 (FIG. 4a) and Hras cells (FIG. 4b) are treated with 100-200 pCi of tracer and incubated for 50 minutes. The cells are measured in a gamma-counter, with the activity normalized to the number of cells used. In both cases, the senescent cells show significantly higher uptake compared to the controls.
The PET-tracer is tested in two different models in vitro. In HCT116 cells senescence is induced by overnight incubation with doxorubicin, followed by 4 days of culture under normal conditions. Senescence in a HRas driven liver progenitor cell line with a doxycycline regulatable p53-specific shRNA. After induction of senescence, both cell lines are incubated with 3.7-7.4 MBq of tracer for 50 minutes. The cells are then washed, counted and measured in a gamma counter. The activity taken up by the cells is analyzed and normalized to a million cells.
In vivo tests are performed in nude mice bearing s.c. HCT116 or HRas driven liver progenitor cells. Mice bearing HCT116 tumors are treated with doxorubicin (10 mg/Kg bodyweight) and imaged 4 days after the onset of senescence. Mice bearing Hras driven tumor are supplied with doxycycline in their drinking water, which has been later removed to induce senescence. After induction of senescence, the tracer is administered i.v. and PET scans are performed. The %ID/cc and tumor-to-muscle ratios are determined through quantitative analysis of the PET data, permitting evaluation of the tracers. In vitro senescent HCT116 cells show an uptake of 11 kBq/1 mio cells, while the uptake in non-senescent control cells is only 4 kBq/1 mio cells. In the HRas driven liver progenitor model senescent cells show an uptake of 192 kBq/1 mio cells and is significantly increased in comparison to non-senescent cells (63 kBq/1 mio cells).
The in vivo tracer uptake in senescent and non-senescent HCT116 tumors (FIG. 5) is 1.7+/−0.7 (n=7) vs 1.1+/−0.4 (n=5) %ID/cc respectively; the respective TMR values are 1.7 and 1.1. In particular, FIG. 5 indicates that tracer 9 shows higher uptake in senescent vs control HCT116 tumors. Both %ID/cc and TMR are higher in senescent vs control tumors. The excised tumors are subjected to ex vivo analyses. Autoradiography confirmed findings, showing higher tracer uptake in the senescent tumor section vs control. Senescent tumor slices also showed higher X-Gal staining than control sections.
In the HRas driven model (FIG. 6), the tracer uptake in senescent tumors is significantly increased to that of control tumors (1.5+/−0.3 (n=16) vs 0.9+/−0.3 (n=11) %ID/cc); the TMR is subject to a similar trend, with respective values of 2.1 and 1.2. FIG. 6 indicates that tracer 9 shows higher uptake in senescent vs control HRas tumors. Both %ID/cc and TMR are higher in senescent vs control tumors. The excised tumors are subjected to ex vivo analyses. Autoradiography confirmed findings, showing higher tracer uptake in the senescent tumor section vs control. Senescent tumor slices also show higher X-Gal staining than control sections. The nonsenescent tumor expresses GFP, which can be visualized with optical imaging.
Induction of senescence is confirmed by ex vivo beta-gal staining and immunohistology of Ki67, Caspase3, HP1γ, p53 and p16 (FIG. 7). FIG. 7 shows immunohistochemical staining from HCT116 (FIG. 7a) and HRas (FIG. 7b) driven tumor models. In both cases, immunohistochemical analysis was performed and reveals an increase of the expected senescence markers HP1γ and p53. The Hras driven tumors also show high expression of ki67. The low abundance of caspase-3 in both tumor models suggests that apoptosis is not a key contributor to the tumors status and confirms assertion of senescence.
1. A compound of the formula:
G-S-L, or a salt thereof
wherein G is
R is H, substituted or unsubstituted C1 to C5 alkyl, substituted or unsubstituted C1 to C10 cycloalkyl, or substituted or unsubstituted C1 to C10 heterocycloalkyl,
* represents the binding site between G and S,
S is
wherein X is independently H, halogen, methyl halogen, OH or SH,
wherein Y is independently C, S, N, or O, with the proviso that at least 3 C-atoms are present,
wherein # represents the binding site between S and L, and wherein L is
wherein Z is a radioactive detectable label, a radioactive therapeutic residue, a chelator coordinating a radioactive detectable label or a chelator coordinating a radioactive therapeutic residue.
2. The compound according to claim 1, wherein G is
3. The compound according to claim 1, wherein
R is H or CH3,
X is independently H or methyl halide,
Y is independently C or N with the proviso that at least 5 C-atoms are present,
R′ is NH,
n is 1, and
R″ is a linear ether having 2 to 5 C-atoms.
R is H,
5. The compound according to claim 1, wherein the compound is
6. The compound according to claim 1, wherein the radioactive detectable label is 11C, 40K, 13N, 15O, 18F, 75Br, 76Br, 82Rb, 68Ga, 64Cu, 62Cu, 123I, 124I, 125I, 131I, 210At, 211At and 111In.
7. The compound according to claim 1, wherein the radioactive therapeutic residue is 32P, 60Co, 64Cu, 89Sr, 90Y, 177Lu, 186Re and 153Sm.
8. A method for detecting senescent cells, comprising the steps of contacting cells suspected to comprise senescent cells with the compound of claim 1 or the salt thereof,
wherein Z is the radioactive detectable label or the chelator coordinating the radioactive detectable label, thereby enriching said compound in senescent cells, if senescent cells are present among said cells, and
detecting the senescent cells having the compound enriched therein.
9. A method for detecting tumor cells, comprising:
contacting cells suspected of comprising tumor cells with the compound of claim 1 of the salt thereof, wherein Z is a radioactive detectable label or the chelator coordinating the radioactive detectable label, thereby enriching the compound in tumor cells, if tumor cells are present among said cells, and
detecting tumor cells having the compound enriched therein.
10. A method for determining the efficiency of cancer treatment, comprising:
contacting cells with the compound of claim 1 or the salt thereof, wherein Z is the radioactive detectable label or the chelator coordinating the radioactive detectable label.
11. A method for determining the efficiency of cancer treatment, the method comprising:
(i) contacting cells of a subject undergoing a cancer treatment with the compound of claim 1 or the salt thereof, wherein Z is the radioactive detectable label or the chelator coordinating the radioactive detectable label, thereby enriching said compound in cancer cells, if present in said cells, and thereby determining a first extent to which cancer cells have enriched therein the compound;
(ii) repeating step (i) after a certain time period during the cancer treatment to determine a second extent to which cancer cells have enriched therein the compound; and (iii) comparing the first and second extent, thereby determining the efficiency of cancer treatment.
12. A method for treating cancer in a subject in need of a treatment, comprising:
administering to the subject a therapeutically effective amount of the compound of claim 1 or the salt thereof, wherein Z is the radioactive therapeutic residue or the chelator coordinating the radioactive therapeutic residue.
13. The method of claim 12, wherein the compound is administered to the subject in a pharmaceutical composition comprising the compound and an inert, non-toxic, pharmaceutically suitable exci
Applicant: Eberhard Karls Universitaet Tuebingen Medizinische Fakultaet (Tuebingen)
Inventors: Jonathan Cotton (Tuebingen), Anna Kuehn (Hechingen), Andreas Maurer (Tuebingen), Bernd Pichler (Scheyern), Klaus Schulze-Osthoff (Tuebingen), Kerstin Fuchs (Steinenbronn), Marcel André Krueger (Reutlingen), Lars Zender (Rottenburg - Wendelsheim)
International Classification: A61K 51/04 (20060101); C07H 17/02 (20060101); C07B 59/00 (20060101); G01N 33/50 (20060101);